BACKGROUND The high incidence and mortality of gastric cancer(GC)pose a significant threat to human life and health,and it has become an important public health challenge in China.Body weight loss is a common complica...BACKGROUND The high incidence and mortality of gastric cancer(GC)pose a significant threat to human life and health,and it has become an important public health challenge in China.Body weight loss is a common complication after surgical treatment in patients with GC and is associated with poor prognosis and GC recurrence.However,current attention to postoperative weight change in GC patients remains insufficient,and the descriptions of postoperative weight change and its influencing factors are also different.AIM To investigate body weight changes in patients with GC within 6 mo after gastrectomy and identify factors that influence dynamic body weight changes.METHODS We conducted a prospective longitudinal study of 121 patients with GC and collected data before(T0)and 1(T1),3(T2),and 6(T3)mo after gastrectomy using a general data questionnaire,psychological distress thermometer,and body weight measurements.The general estimation equation(GEE)was used to analyze the dynamic trends of body weight changes and factors that influence body weight changes in patients with GC within 6 mo of gastrectomy.RESULTS The median weight loss at T1,T2,and T3 was 7.29%(2.84%,9.40%),11.11%(7.64%,14.91%),and 14.75%(8.80%,19.84%),respectively.The GEE results showed that preoperative body mass index(BMI),significant psychological distress,religious beliefs,and sex were risk factors for weight loss in patients with GC within 6 mo after gastrectomy(P<0.05).Compared with preoperative low-weight patients,preoperative obese patients were more likely to have weight loss(β=14.685,P<0.001).Furthermore,patients with significant psychological distress were more likely to lose weight than those without(β=2.490,P<0.001),and religious patients were less likely to lose weight 6 mo after gastrectomy than those without religious beliefs(β=-6.844,P=0.001).Compared to female patients,male patients were more likely to experience weight loss 6 mo after gastrectomy(β=4.262,P=0.038).CONCLUSION Male patients with GC with high preoperative BMI,significant psychological distress,and no religious beliefs are more likely to lose weight after gastrectomy.展开更多
AIM To clarify the mechanisms of connexin 32 (Cx32) downregulation by potential transcriptional factors (TFs) in Helicobacter pylori (H. pylori)-associated gastric carcinogenesis. METHODS Approximately 25 specimens at...AIM To clarify the mechanisms of connexin 32 (Cx32) downregulation by potential transcriptional factors (TFs) in Helicobacter pylori (H. pylori)-associated gastric carcinogenesis. METHODS Approximately 25 specimens at each developmental stage of gastric carcinogenesis [non-atrophic gastritis, chronic atrophic gastritis, intestinal metaplasia, dysplasia and gastric carcinoma (GC)] with H. pylori infection [H. pylori (+)] and 25 normal gastric mucosa (NGM) without H. pylori infection [H. pylori (-)] were collected. After transcriptional factor array analysis, the Cx32 and PBX1 expression levels of H. pylori-infected tissues from the developmental stages of GC and NGM with no H. pylori infection were measured by real-time polymerase chain reaction (RT-PCR) and Western blot analysis. Regarding H. pylori-infected animal models, the Cx32 and PBX1 mRNA expression levels and correlation between the gastric mucosa from 10 Mongolian gerbils with long-term H. pylori colonization and 10 controls were analyzed. PBX1 and Cx32 mRNA and protein levels were further studied under the H. pylori-infected condition as well as PBX1 overexpression and knockdown conditions in vitro. RESULTS Incremental PBX1 was first detected by TF microarray in H. pylori-related gastric carcinogenesis. The identical trend of PBX1 and Cx32 expression was confirmed in the developmental stages of H. pylori-related clinical specimens. The negative correlation of PBX1 and Cx32 was confirmed in H. pylori-infected Mongolian gerbils. Furthermore, decreased PBX1 expression was detected in the normal gastric epithelial cell line GES-1 with H. pylori infection. Enforced overexpression or RNAi-mediated knockdown of PBX1 contributed to the diminished or restored Cx32 expression in GES-1 and the gastric carcinoma cell line BGC823, respectively. Finally, dual-luciferase reporter assay in HEK293T cells showed that Cx32 promoter activity decreased by 30% after PBX1 vector co-transfection, indicating PBX1 as a transcriptional downregulator of Cx32 by directly binding to its promoters. ONCLUSION PBX1 is one of the determinants in the Cx32 promoter targeting site, preventing further damage of gap junction protein in H. pylori-associated gastric carcinogenesis.展开更多
Background:The supine position is the most common birth position adopted in China,but the World Health Organization recommends non-supine positions for delivery.The handsand-knees position shows several advantages,suc...Background:The supine position is the most common birth position adopted in China,but the World Health Organization recommends non-supine positions for delivery.The handsand-knees position shows several advantages,such as wide pelvic diameter and easy fetal rotation during delivery.Small trials conducted in China in 2011 revealed that the handsand-knees position resulted in improved maternal and neonate outcomes than those in the supine position.However,a comprehensive study must be conducted before the handsand-knees position can be introduced into clinical practice.Hence,we conducted this multicenter trial to comprehensively examine the benefits of the hands-and-knees position over the supine position during delivery.Methods:Our clinical study was conducted in 11 hospitals in China from May to December 2012.A total of 446 pregnant women who gave birth in the hands-and-knees position were assigned into the experimental group,and 440 women who gave birth in the supine position were classified into the control group.Episiotomy rate was evaluated as the primary outcome,and perineum laceration degree was considered the secondary outcome.Results:Women in the experimental group achieved lower rates of episiotomy and higher rates of intact perineum and first-degree perineum lacerations compared with those in the control.Postpartum bleeding amount,neonatal asphyxia,and APGAR scores at 1 and 5 min were not significantly different between the two groups.Conclusions:This study proves that women who delivered in the hands-and-knees position achieved low rates of episiotomy and intact perineum.Moreover,the rates of neonatal asphyxia and postpartum bleeding did not increase.Pregnant patients who prefer to adopt the hands-and-knees position should be assisted in assuming such position during delivery.展开更多
AIM:To identify genes potentially involved in Helicobacter pylori(H.pylori)-induced gastric carcinogenesis.METHODS:GES-1 cells were co-cultured with H.pylori strains isolated from patients with gastric carcinoma(GC,n ...AIM:To identify genes potentially involved in Helicobacter pylori(H.pylori)-induced gastric carcinogenesis.METHODS:GES-1 cells were co-cultured with H.pylori strains isolated from patients with gastric carcinoma(GC,n = 10) or chronic gastritis(CG,n = 10) for in vitro proliferation and apoptosis assays to identify the most and least virulent strains.These two strains were cagA-genotyped and used for further in vivo carcinogenic virulence assays by infecting Mongolian gerbils for 52 wk,respectively;a broth free of H.pylori was lavaged as control.Genomic profiles of GES-1 cells cocultured with the most and least virulent strains were determined by microarray analysis.The most differentially expressed genes were further verified using quantitative real-time polymerase chain reaction in GES-1cells infected with the most and least virulent strains,and by immunohistochemistry in H.pylori positive CG,precancerous diseases,and GC biopsy specimens in an independent experiment.RESULTS:GC-derived H.pylori strains induced a potent proliferative effect in GES-1 cells in co-culture,whereas CG-derived strains did not.The most(from a GC patient) and least(from a CG patient) virulent strains were cagA-positive and negative,respectively.At week 52,CG,atrophy,metaplasia,dysplasia,and GC were observed in 90.0%,80.0%,80.0%,90%,and 60.0%,respectively,of the animals lavaged with the most virulent strain.However,only mild CG was observed in 90% of the animals lavaged with the least virulent strain.On microarray analysis,800 differentially expressed genes(49 up-and 751 down-regulated),involving those associated with cell cycle regulation,cell apoptosis,cytoskeleton,immune response,and substance and energy metabolisms,were identified in cells co-cultured with the most virulent strain as compared with those co-cultured with the least virulent strain.The six most differentially expressed genes(with a betweenness centrality of 0.1-0.2) were identified among the significant differential gene profile network,including JUN,KRAS,BRCA1,SMAD2,TRAF1,and HDAC6.Quantitative real-time polymerase chain reaction analyses verified that HDAC6 and TRFA1 mRNA expressions were significantly more up-regulated in GES-1 cells cocultured with the most virulent strain than in those cocultured with the least virulent strain.Immunohistochemistry of gastric mucosal specimens from H.pyloripositive patients with CG,intestinal metaplasia(IM),dysplasia,and GC showed that moderately positive and strongly positive HDAC6 expression was detected in 21.7% of CG patients,30.0% of IM patients,54.5% of dysplasia patients,and 77.8% of GC patients(P < 0.001).The up-regulation of TRAF1 expressions was detected in 34.8%,53.3%,72.7%,and 88.9% specimens of CG,IM,dysplasia,and GC,respectively(P < 0.001).CONCLUSION:The overexpression of HDAC6 and TRAF1 in GES-1 cells co-cultured with the GC-derived strain and in H.pylori-positive dysplasia and GC suggests that HDAC6 and TRAF1 may be involved in H.pyloriinduced gastric carcinogenesis.展开更多
Objective: To investigate the azole susceptibility of Candida albicans(C.albicans)from vulvovaginal candidosis patients and to analyze the relationship between ERG11 gene mutations in these isolates and azole resistan...Objective: To investigate the azole susceptibility of Candida albicans(C.albicans)from vulvovaginal candidosis patients and to analyze the relationship between ERG11 gene mutations in these isolates and azole resistance.Methods: Three hundred and two clinical isolates of Candida species were collected.Azole susceptibility was tested in vitro in microdilution studies. The ERG11 genes of 17 isolates of C. albicans(2 susceptibles, 5 dose-dependent resistants and 10 resistants) were amplified and sequenced.Results: Of the 302 isolates collected, 70.2% were C. albicans, of which 8.5%, 3.8% and4.2% were resistant to fluconazole, itraconazole and voriconazole, respectively. In total,27 missense mutations were detected in ERG11 genes from resistant/susceptible dosedependent isolates. Among them, Y132 H, A114 S, and Y257 H substitutions were most prevalent and were known to cause fluconazole resistance. G464 S and F72 S also have been proved to cause fluconazole resistance. Two novel substitutions(T285A, S457P) in hotspot regions were identified.Conclusions: Twenty seven mutations in the ERG11 gene were identified in azoleresistant C. albicans isolates, which indicated a possible relation with the increase in resistance to azole drugs and the recurrence of vulvovaginal candidosis. The relationship of two novel substitutions(T285A, S457P) with fluconazole resistance needs to be further verified by site-directed mutagenesis.展开更多
Molting and metamorphosis are important physiological processes in insects that are tightly controlled by ecdysone receptor(EcR)through the 20-hydroxyecdysone(20E)signaling pathway.EcR is a steroid nuclear receptor(SR...Molting and metamorphosis are important physiological processes in insects that are tightly controlled by ecdysone receptor(EcR)through the 20-hydroxyecdysone(20E)signaling pathway.EcR is a steroid nuclear receptor(SR).Several FK506-binding proteins(FKBPs)have been identified from the mammal SR complex,and are thought to be involved in the subcellular trafficking of SR.However,their roles in insects are poorly understood.To explore whether FKBPs are involved in insect molting or metamorphosis,we injected an FKBP inhibitor(FK506)into a lepidopteran insect,Spodoptera litura,and found that molting was inhibited in 61.11%of the larvae,and that the time for larvae to pupate was significantly extended.A total of 10 FKBP genes were identified from the genome of s.litura and were clustered into 2 distinct groups,according to their subcellular localization,with FKBP13 and FKBP14 belonging to the endoplasmic reticulum(ER)group and with the other members belonging to the cytoplasmic(Cy)group.All the CyFKBPs were significantly upregulated in the prepupal or pupal stages,with the opposite being observed for the ER group members.FK506 completely blocked the transfer of EcR to the nucleus under 20E induction,and significantly downregulated the transcriptional expression of many 20E signaling genes.A similar phenomenon was observed after RNA interference of2 CyFKBPs(FKBP45 and FKBP12b),but not for FKBP13.Taken together,our data indicate that the cytoplasmic FKBPs,especially FKBP45 and FKBP12b,mediate the nuclear localization of EcR,thereby regulating the 20E signaling and ultimately affecting molting and metamorphosis in insects.展开更多
Background and Aims:Chronic hepatitis B(CHB)can cause liver fibrosis and lead to cirrhosis and cancer.As the effectiveness of antiviral therapy to reverse liver fibrosis is limited,We aimed to evaluate the effect of A...Background and Aims:Chronic hepatitis B(CHB)can cause liver fibrosis and lead to cirrhosis and cancer.As the effectiveness of antiviral therapy to reverse liver fibrosis is limited,We aimed to evaluate the effect of An-Luo-Hua-Xian pill(ALHX)on fibrosis regression in CHB patients treated with entecavir(ETV).Methods:Treatment-naïve patients with CHB were randomly treated with ETV alone or combined with ALHX(ETV+ALHX)between October 1,2013 and December 31,2020.Demographic,laboratory,and liver histology data before and after 78 weeks of treatment were collected.The Ishak fibrosis score(F)was used and fibrosis regression required a decrease in F of≥1 after treatment.Results:A total of 780 patients were enrolled,and 394 with a second liver biopsy after treatment were included in the per-protocol population,132 in ETV group and 262 in ETV+ALHX group.After 78 weeks of treatment,the fibrosis regression rate in the ETV+ALHX group was significantly higher than that of the ETV group at baseline F≥3 patients:124/211(58.8%)vs.45/98(45.9%),p=0.035.The percentage of patients with a decreased liver stiffness measurement(LSM)was higher in the ETV+ALHX group:156/211(73.9%)vs.62/98(63.%),p=0.056.Logistic regression analysis showed that ETV combined with ALHX was associated with fibrosis regression[odds ratio(OR)=1.94,p=0.018],and a family history of hepatocellular carcinoma was on the contrary.(OR=0.41,p=0.031).Conclusions:ETV combined with ALHX increased liver fibrosis regression in CHB patients.展开更多
Ecdysone receptor （EcR） and ultraspiracle （USP） form heterodimers to mediate ecdysteroid signaling during molting and metamorphosis. Various EcR/USP heterodimers have been reported. However, it is unclear what kin...Ecdysone receptor （EcR） and ultraspiracle （USP） form heterodimers to mediate ecdysteroid signaling during molting and metamorphosis. Various EcR/USP heterodimers have been reported. However, it is unclear what kind of EcR/USP combination is adopted by lepidopteran insects during the larval-pupal metamorphosis and whether the EcR/USP heterodimer varies among different tissues. To address these questions, two isoforms of each EcR and USP were cloned from the common cutworm, their messenger RNA expression patterns were examined by real-time quantitative polymerase chain reaction in different tissues during the larval-pupal metamorphosis and in the midgut in response to hormonal induction. Furthermore, their subcellular localization and protein-protein interaction were explored by transient expression and far-western blotting, respectively. All the four genes were significantly up-regulated in prepuae and/or pupae. The expression profiles of EcRB1 and USP1 were nearly identical to each other in the epidermis, fat body and midgut, and a similar situation also applied to EcRA and USP2. The three genes responded to 20-hydroxyecdysone （20E） induction except for USP2, and USP1 could be up-regulated by both 20E and juvenile hormone. The four proteins mainly localized in the nucleus and the nuclear localization was promoted by 20E. The protein-protein interaction between each EcR and USP was found in vitro. These results suggest that two types of EcR/USP heterodimer （EeRA/USP2 and EcRB 1/USP1） may exist simultaneously in the common cutworm, and the latter should play more important roles during the larval-pupal metamorphosis. In addition, the types of EcR/USP heterodimer do not vary in the tissues which undergo histolysis and regeneration during metamorphosis.展开更多
Chlorpyrifos （CPF） is a broad-spectrum organophosphate insecticide. Glu- tathione S-transferases （GSTs） in insects are a family of detoxification enzymes and they play critical roles in CPF detoxification. Spodopt...Chlorpyrifos （CPF） is a broad-spectrum organophosphate insecticide. Glu- tathione S-transferases （GSTs） in insects are a family of detoxification enzymes and they play critical roles in CPF detoxification. Spodoptera litura is one of the most destructive agricultural pests in tropical and subtropical areas in the world. In this study, 37 Slgsts from 46 unique transcripts of gsts in S. litura transcriptome data, including eight pre- viously reported GSTs, were identified and their expression patterns in susceptible and 12-generation-CPF-treated strains were analyzed to understand the roles of these Slgsts in sublethal doses of CPF tolerance. The results indicate that the members of the S. litura GST superfamily could be distinguished into three major groups： one group, includ- ing six cytosolic Slgsts （SIGSTel, SIGSTe3, SIGSTelO, SIGSTe15, SIGSTo2 and SIGSTs5） and two microsomal Slgsts （SIMGSTI-2 and SIMGST1-3）, was directly responsible for CPF induction in both 12-generation-treated and susceptible strains; the second group, including three cytosolic Slgsts （SlGSTe13, SIGSTtl and SIGSTzl） and one microsomal Slgst （SIMGSTI-1）, was induced only in the 12-generation-treated strain; the third group, including eight cytosolic Slgsts （two epsilon, three delta, one omega, one zeta and one un- classified Slgst）, was expressed 1.52-5.15-fold higher in the 12-generation-treated strain than in the susceptible strain.展开更多
Metamorphosis is one of the most important physiological processes in insects.It is regulated by a serial of ecdysone cascade genes.Recently,lots o f microRNAs(miRNAs)were investigated in insects;however,their functio...Metamorphosis is one of the most important physiological processes in insects.It is regulated by a serial of ecdysone cascade genes.Recently,lots o f microRNAs(miRNAs)were investigated in insects;however,their function in metamorphosis is largely unknown.In the present study,the dynamics of a small RNA population was investigated by RNA sequencing from the midgut of a lepidopteran pest Spodoptera litura during larval-pupal metamorphosis.A total of 101 miRNAs were identified,and 75 miRNAs were differentially expressed during the metamorphic process.The relationship between these differentially expressed miRNAs and 12 ecdysone cascade genes was analyzed by four classical software programs,and a multiple-to-multiple regulatory network was found to exist between these miRNAs and their targets.Among them,miR-14-3p and its two targets(EcR and E75)were chosen for further validation.MiR-14-3p had higher expression level in the 6 th instar larvae as compared with either the prepupae or pupae,which was opposite to that of both EcR and E75,two ecdysone cascade genes.Luciferase reporter assay confirmed that both EcR and E75 were regulated by miR-14-3p.Interestingly,the 3'untranslated regions are nearly identical to each other among different transcript variants of the ecdysone cascade genes,including EcR,USP,E75,E74,E78,E93,Hr3,Hr4,Hr39,K rhl and F tzfl.Thus,different transcript variants of one ecdysone cascade gene could be regulated by the same mi RNA.The above data suggest that the ecdysone signaling pathway is under the tight control of miRNA.These findings expand our understanding of the mechanism of insect metamorphosis and may also provide a novel possibility for the control of pest insects in the future.展开更多
Silkworm mutants are valuable resources for both transgenic breeding and gene discovery. PiggyBac-based random insertional mutagenesis has been widely used in gene functional studies. In order to discover genes involv...Silkworm mutants are valuable resources for both transgenic breeding and gene discovery. PiggyBac-based random insertional mutagenesis has been widely used in gene functional studies. In order to discover genes involved in silk synthesis, a piggyBac-based random insertional library was constructed using Bombyx mori, and the mutants with abnormal cocoon were particularly screened. By this means, a “thin cocoon” mutant was identified. This mutant revealed thinner cocoon shell and shorter posterior silk gland (PSG) compared with the wild type. The messenger RNA (mRNA) levels of all the three fibroin genes, including Fib-H, Fib-L and P25, were significantly down-regulated in the PSG of mutants. Four piggyBac insertion sites were identified in Aquaporin (AQP), Longitudinals lacking protein-like {Lola), Glutamyl aminopeptidase-like (GluAP) and Loc101744460. The mRNA levels of all the four genes were significantly altered in the silk gland of mutants. In particular, the mRNA amount of AQP, a gene responsible for the regulation of osmotic pressure, decreased dramatically immediately prior to the spinning stage in the anterior silk gland of mutants. The identification of the genes disrupted in the “thin cocoon” mutant in this study provided useful information for understanding silk production and transgenic breeding of silkworms in the future.展开更多
Objective To establish a reverse-phase liquid chromatography method for the determination of seven alkaloids(magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine)in Fufang Zhenzhu Tiao...Objective To establish a reverse-phase liquid chromatography method for the determination of seven alkaloids(magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine)in Fufang Zhenzhu Tiaozhi Capsule.Methods Chromatography was performed on a Dionex Acclaim C_(18)column(250 mm×4.6 mm,5.0μm)at 30℃.The mobile phase was composed of acetonitrile-potassium dihydrogen phosphate solution(0.015 mol/L,40:60,including 1.7 g/L sodium dodecyl sulfate and phosphoric acid used to regulate pH value to 3.0),with a flow rate of 1.0 mL/min.The detection wavelength was 270 nm.Results The calibration curves of magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine were linear in the range of 1.07-10.65,0.78-7.55,0.75-7.50,1.60-15.95,2.69-26.85,2.31-23.10,and 6.04-60.40 mg/mL.The average recoveries of magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine were 101.0%,101.2%,100.1%,100.0%,100.1%,101.1%,and 99.7%,respectively.Conclusion The method could be used for the quantitative determination of the preparation.展开更多
Juvenile hormone （JH） is one of the key insect hormones that regulate metamorphosis. Juvenile hormone diol kinase （JHDK） is an enzyme involved in JH metabolism and catalyzes JH diol to form a polar end product, JH...Juvenile hormone （JH） is one of the key insect hormones that regulate metamorphosis. Juvenile hormone diol kinase （JHDK） is an enzyme involved in JH metabolism and catalyzes JH diol to form a polar end product, JH diol phosphate that has no JH activity. In this study, a JHDK complementary DNA （cDNA） was cloned from Spodoptera litura and the structure and expression of the gene was characterized. The cDNA was 714 base pairs in length and encoded a protein of 183 amino acids with a molecular mass of 21 kDa and an isoelectric point of 4.55. Based on the structure, three putative calcium binding motifs and guanosine triphosphate-binding motifs were predicted in the protein. Modeling of the 3-D structure showed that the protein consisted of eight α-helixes linked with loops, with no β-sheets. The gene was expressed in the epidermis, fat body and midgut of fifth and sixth instar larvae. The expression level in the epidermis was lower than in the fat body and midgut. The gene was expressed at higher levels at the early stages than in the later stages of fifth and sixth instar midgut and fat body. The results suggest that this gene may be involved in the regulation of the JH titer in larvae ofS. litura.展开更多
文摘BACKGROUND The high incidence and mortality of gastric cancer(GC)pose a significant threat to human life and health,and it has become an important public health challenge in China.Body weight loss is a common complication after surgical treatment in patients with GC and is associated with poor prognosis and GC recurrence.However,current attention to postoperative weight change in GC patients remains insufficient,and the descriptions of postoperative weight change and its influencing factors are also different.AIM To investigate body weight changes in patients with GC within 6 mo after gastrectomy and identify factors that influence dynamic body weight changes.METHODS We conducted a prospective longitudinal study of 121 patients with GC and collected data before(T0)and 1(T1),3(T2),and 6(T3)mo after gastrectomy using a general data questionnaire,psychological distress thermometer,and body weight measurements.The general estimation equation(GEE)was used to analyze the dynamic trends of body weight changes and factors that influence body weight changes in patients with GC within 6 mo of gastrectomy.RESULTS The median weight loss at T1,T2,and T3 was 7.29%(2.84%,9.40%),11.11%(7.64%,14.91%),and 14.75%(8.80%,19.84%),respectively.The GEE results showed that preoperative body mass index(BMI),significant psychological distress,religious beliefs,and sex were risk factors for weight loss in patients with GC within 6 mo after gastrectomy(P<0.05).Compared with preoperative low-weight patients,preoperative obese patients were more likely to have weight loss(β=14.685,P<0.001).Furthermore,patients with significant psychological distress were more likely to lose weight than those without(β=2.490,P<0.001),and religious patients were less likely to lose weight 6 mo after gastrectomy than those without religious beliefs(β=-6.844,P=0.001).Compared to female patients,male patients were more likely to experience weight loss 6 mo after gastrectomy(β=4.262,P=0.038).CONCLUSION Male patients with GC with high preoperative BMI,significant psychological distress,and no religious beliefs are more likely to lose weight after gastrectomy.
基金Supported by The National Natural Science Foundation of China,No.81172301Changsha Municipal Science and Technology Project,No.K1106036-31
文摘AIM: To explore the mechanism of abnormal Connexin (Cx) 32 and Cx43 expression in the gastric mucosa after Helicobacter pylori (H. pylori) infection.
基金Supported by the New Xiangya Talent Project of the Third Xiangya Hospital of Central South University,No.20150310
文摘AIM To clarify the mechanisms of connexin 32 (Cx32) downregulation by potential transcriptional factors (TFs) in Helicobacter pylori (H. pylori)-associated gastric carcinogenesis. METHODS Approximately 25 specimens at each developmental stage of gastric carcinogenesis [non-atrophic gastritis, chronic atrophic gastritis, intestinal metaplasia, dysplasia and gastric carcinoma (GC)] with H. pylori infection [H. pylori (+)] and 25 normal gastric mucosa (NGM) without H. pylori infection [H. pylori (-)] were collected. After transcriptional factor array analysis, the Cx32 and PBX1 expression levels of H. pylori-infected tissues from the developmental stages of GC and NGM with no H. pylori infection were measured by real-time polymerase chain reaction (RT-PCR) and Western blot analysis. Regarding H. pylori-infected animal models, the Cx32 and PBX1 mRNA expression levels and correlation between the gastric mucosa from 10 Mongolian gerbils with long-term H. pylori colonization and 10 controls were analyzed. PBX1 and Cx32 mRNA and protein levels were further studied under the H. pylori-infected condition as well as PBX1 overexpression and knockdown conditions in vitro. RESULTS Incremental PBX1 was first detected by TF microarray in H. pylori-related gastric carcinogenesis. The identical trend of PBX1 and Cx32 expression was confirmed in the developmental stages of H. pylori-related clinical specimens. The negative correlation of PBX1 and Cx32 was confirmed in H. pylori-infected Mongolian gerbils. Furthermore, decreased PBX1 expression was detected in the normal gastric epithelial cell line GES-1 with H. pylori infection. Enforced overexpression or RNAi-mediated knockdown of PBX1 contributed to the diminished or restored Cx32 expression in GES-1 and the gastric carcinoma cell line BGC823, respectively. Finally, dual-luciferase reporter assay in HEK293T cells showed that Cx32 promoter activity decreased by 30% after PBX1 vector co-transfection, indicating PBX1 as a transcriptional downregulator of Cx32 by directly binding to its promoters. ONCLUSION PBX1 is one of the determinants in the Cx32 promoter targeting site, preventing further damage of gap junction protein in H. pylori-associated gastric carcinogenesis.
文摘Background:The supine position is the most common birth position adopted in China,but the World Health Organization recommends non-supine positions for delivery.The handsand-knees position shows several advantages,such as wide pelvic diameter and easy fetal rotation during delivery.Small trials conducted in China in 2011 revealed that the handsand-knees position resulted in improved maternal and neonate outcomes than those in the supine position.However,a comprehensive study must be conducted before the handsand-knees position can be introduced into clinical practice.Hence,we conducted this multicenter trial to comprehensively examine the benefits of the hands-and-knees position over the supine position during delivery.Methods:Our clinical study was conducted in 11 hospitals in China from May to December 2012.A total of 446 pregnant women who gave birth in the hands-and-knees position were assigned into the experimental group,and 440 women who gave birth in the supine position were classified into the control group.Episiotomy rate was evaluated as the primary outcome,and perineum laceration degree was considered the secondary outcome.Results:Women in the experimental group achieved lower rates of episiotomy and higher rates of intact perineum and first-degree perineum lacerations compared with those in the control.Postpartum bleeding amount,neonatal asphyxia,and APGAR scores at 1 and 5 min were not significantly different between the two groups.Conclusions:This study proves that women who delivered in the hands-and-knees position achieved low rates of episiotomy and intact perineum.Moreover,the rates of neonatal asphyxia and postpartum bleeding did not increase.Pregnant patients who prefer to adopt the hands-and-knees position should be assisted in assuming such position during delivery.
基金Supported by Grants from the Department of Science and Technology,No. 2011FJ6087the Natural Science Foundation of Hunan Province,China,No. 10JJ5035
文摘AIM:To identify genes potentially involved in Helicobacter pylori(H.pylori)-induced gastric carcinogenesis.METHODS:GES-1 cells were co-cultured with H.pylori strains isolated from patients with gastric carcinoma(GC,n = 10) or chronic gastritis(CG,n = 10) for in vitro proliferation and apoptosis assays to identify the most and least virulent strains.These two strains were cagA-genotyped and used for further in vivo carcinogenic virulence assays by infecting Mongolian gerbils for 52 wk,respectively;a broth free of H.pylori was lavaged as control.Genomic profiles of GES-1 cells cocultured with the most and least virulent strains were determined by microarray analysis.The most differentially expressed genes were further verified using quantitative real-time polymerase chain reaction in GES-1cells infected with the most and least virulent strains,and by immunohistochemistry in H.pylori positive CG,precancerous diseases,and GC biopsy specimens in an independent experiment.RESULTS:GC-derived H.pylori strains induced a potent proliferative effect in GES-1 cells in co-culture,whereas CG-derived strains did not.The most(from a GC patient) and least(from a CG patient) virulent strains were cagA-positive and negative,respectively.At week 52,CG,atrophy,metaplasia,dysplasia,and GC were observed in 90.0%,80.0%,80.0%,90%,and 60.0%,respectively,of the animals lavaged with the most virulent strain.However,only mild CG was observed in 90% of the animals lavaged with the least virulent strain.On microarray analysis,800 differentially expressed genes(49 up-and 751 down-regulated),involving those associated with cell cycle regulation,cell apoptosis,cytoskeleton,immune response,and substance and energy metabolisms,were identified in cells co-cultured with the most virulent strain as compared with those co-cultured with the least virulent strain.The six most differentially expressed genes(with a betweenness centrality of 0.1-0.2) were identified among the significant differential gene profile network,including JUN,KRAS,BRCA1,SMAD2,TRAF1,and HDAC6.Quantitative real-time polymerase chain reaction analyses verified that HDAC6 and TRFA1 mRNA expressions were significantly more up-regulated in GES-1 cells cocultured with the most virulent strain than in those cocultured with the least virulent strain.Immunohistochemistry of gastric mucosal specimens from H.pyloripositive patients with CG,intestinal metaplasia(IM),dysplasia,and GC showed that moderately positive and strongly positive HDAC6 expression was detected in 21.7% of CG patients,30.0% of IM patients,54.5% of dysplasia patients,and 77.8% of GC patients(P < 0.001).The up-regulation of TRAF1 expressions was detected in 34.8%,53.3%,72.7%,and 88.9% specimens of CG,IM,dysplasia,and GC,respectively(P < 0.001).CONCLUSION:The overexpression of HDAC6 and TRAF1 in GES-1 cells co-cultured with the GC-derived strain and in H.pylori-positive dysplasia and GC suggests that HDAC6 and TRAF1 may be involved in H.pyloriinduced gastric carcinogenesis.
基金Supported by the National Natural Science Foundation of China(No.31072125 and 31272549)by a National Science and Technology Project (China) grant (2008ZX-10004-011)
文摘Objective: To investigate the azole susceptibility of Candida albicans(C.albicans)from vulvovaginal candidosis patients and to analyze the relationship between ERG11 gene mutations in these isolates and azole resistance.Methods: Three hundred and two clinical isolates of Candida species were collected.Azole susceptibility was tested in vitro in microdilution studies. The ERG11 genes of 17 isolates of C. albicans(2 susceptibles, 5 dose-dependent resistants and 10 resistants) were amplified and sequenced.Results: Of the 302 isolates collected, 70.2% were C. albicans, of which 8.5%, 3.8% and4.2% were resistant to fluconazole, itraconazole and voriconazole, respectively. In total,27 missense mutations were detected in ERG11 genes from resistant/susceptible dosedependent isolates. Among them, Y132 H, A114 S, and Y257 H substitutions were most prevalent and were known to cause fluconazole resistance. G464 S and F72 S also have been proved to cause fluconazole resistance. Two novel substitutions(T285A, S457P) in hotspot regions were identified.Conclusions: Twenty seven mutations in the ERG11 gene were identified in azoleresistant C. albicans isolates, which indicated a possible relation with the increase in resistance to azole drugs and the recurrence of vulvovaginal candidosis. The relationship of two novel substitutions(T285A, S457P) with fluconazole resistance needs to be further verified by site-directed mutagenesis.
基金supported by grants from the National Natural Science Foundation of China(grant no.32272523)the Natural Science Foundation of Guangdong Province,China(grant no.2023A1515010178)the Laboratory of Lingnan Modern Agriculture Project(grant no.NT2021003).
文摘Molting and metamorphosis are important physiological processes in insects that are tightly controlled by ecdysone receptor(EcR)through the 20-hydroxyecdysone(20E)signaling pathway.EcR is a steroid nuclear receptor(SR).Several FK506-binding proteins(FKBPs)have been identified from the mammal SR complex,and are thought to be involved in the subcellular trafficking of SR.However,their roles in insects are poorly understood.To explore whether FKBPs are involved in insect molting or metamorphosis,we injected an FKBP inhibitor(FK506)into a lepidopteran insect,Spodoptera litura,and found that molting was inhibited in 61.11%of the larvae,and that the time for larvae to pupate was significantly extended.A total of 10 FKBP genes were identified from the genome of s.litura and were clustered into 2 distinct groups,according to their subcellular localization,with FKBP13 and FKBP14 belonging to the endoplasmic reticulum(ER)group and with the other members belonging to the cytoplasmic(Cy)group.All the CyFKBPs were significantly upregulated in the prepupal or pupal stages,with the opposite being observed for the ER group members.FK506 completely blocked the transfer of EcR to the nucleus under 20E induction,and significantly downregulated the transcriptional expression of many 20E signaling genes.A similar phenomenon was observed after RNA interference of2 CyFKBPs(FKBP45 and FKBP12b),but not for FKBP13.Taken together,our data indicate that the cytoplasmic FKBPs,especially FKBP45 and FKBP12b,mediate the nuclear localization of EcR,thereby regulating the 20E signaling and ultimately affecting molting and metamorphosis in insects.
基金supported by National Science and Technology Major Project (2013ZX10002005 and 2017ZX10203202).
文摘Background and Aims:Chronic hepatitis B(CHB)can cause liver fibrosis and lead to cirrhosis and cancer.As the effectiveness of antiviral therapy to reverse liver fibrosis is limited,We aimed to evaluate the effect of An-Luo-Hua-Xian pill(ALHX)on fibrosis regression in CHB patients treated with entecavir(ETV).Methods:Treatment-naïve patients with CHB were randomly treated with ETV alone or combined with ALHX(ETV+ALHX)between October 1,2013 and December 31,2020.Demographic,laboratory,and liver histology data before and after 78 weeks of treatment were collected.The Ishak fibrosis score(F)was used and fibrosis regression required a decrease in F of≥1 after treatment.Results:A total of 780 patients were enrolled,and 394 with a second liver biopsy after treatment were included in the per-protocol population,132 in ETV group and 262 in ETV+ALHX group.After 78 weeks of treatment,the fibrosis regression rate in the ETV+ALHX group was significantly higher than that of the ETV group at baseline F≥3 patients:124/211(58.8%)vs.45/98(45.9%),p=0.035.The percentage of patients with a decreased liver stiffness measurement(LSM)was higher in the ETV+ALHX group:156/211(73.9%)vs.62/98(63.%),p=0.056.Logistic regression analysis showed that ETV combined with ALHX was associated with fibrosis regression[odds ratio(OR)=1.94,p=0.018],and a family history of hepatocellular carcinoma was on the contrary.(OR=0.41,p=0.031).Conclusions:ETV combined with ALHX increased liver fibrosis regression in CHB patients.
基金Acknowledgments The research was supported by the grants from National Natural Science Foundation of China （Grant No. 31172154） and the National Basic Research Program of China （973 Program, No. 2012CB114101）.
文摘Ecdysone receptor （EcR） and ultraspiracle （USP） form heterodimers to mediate ecdysteroid signaling during molting and metamorphosis. Various EcR/USP heterodimers have been reported. However, it is unclear what kind of EcR/USP combination is adopted by lepidopteran insects during the larval-pupal metamorphosis and whether the EcR/USP heterodimer varies among different tissues. To address these questions, two isoforms of each EcR and USP were cloned from the common cutworm, their messenger RNA expression patterns were examined by real-time quantitative polymerase chain reaction in different tissues during the larval-pupal metamorphosis and in the midgut in response to hormonal induction. Furthermore, their subcellular localization and protein-protein interaction were explored by transient expression and far-western blotting, respectively. All the four genes were significantly up-regulated in prepuae and/or pupae. The expression profiles of EcRB1 and USP1 were nearly identical to each other in the epidermis, fat body and midgut, and a similar situation also applied to EcRA and USP2. The three genes responded to 20-hydroxyecdysone （20E） induction except for USP2, and USP1 could be up-regulated by both 20E and juvenile hormone. The four proteins mainly localized in the nucleus and the nuclear localization was promoted by 20E. The protein-protein interaction between each EcR and USP was found in vitro. These results suggest that two types of EcR/USP heterodimer （EeRA/USP2 and EcRB 1/USP1） may exist simultaneously in the common cutworm, and the latter should play more important roles during the larval-pupal metamorphosis. In addition, the types of EcR/USP heterodimer do not vary in the tissues which undergo histolysis and regeneration during metamorphosis.
文摘Chlorpyrifos （CPF） is a broad-spectrum organophosphate insecticide. Glu- tathione S-transferases （GSTs） in insects are a family of detoxification enzymes and they play critical roles in CPF detoxification. Spodoptera litura is one of the most destructive agricultural pests in tropical and subtropical areas in the world. In this study, 37 Slgsts from 46 unique transcripts of gsts in S. litura transcriptome data, including eight pre- viously reported GSTs, were identified and their expression patterns in susceptible and 12-generation-CPF-treated strains were analyzed to understand the roles of these Slgsts in sublethal doses of CPF tolerance. The results indicate that the members of the S. litura GST superfamily could be distinguished into three major groups： one group, includ- ing six cytosolic Slgsts （SIGSTel, SIGSTe3, SIGSTelO, SIGSTe15, SIGSTo2 and SIGSTs5） and two microsomal Slgsts （SIMGSTI-2 and SIMGST1-3）, was directly responsible for CPF induction in both 12-generation-treated and susceptible strains; the second group, including three cytosolic Slgsts （SlGSTe13, SIGSTtl and SIGSTzl） and one microsomal Slgst （SIMGSTI-1）, was induced only in the 12-generation-treated strain; the third group, including eight cytosolic Slgsts （two epsilon, three delta, one omega, one zeta and one un- classified Slgst）, was expressed 1.52-5.15-fold higher in the 12-generation-treated strain than in the susceptible strain.
基金grants from the National Natural Science Foundation of China(Grant No.31672357 and 31872285)。
文摘Metamorphosis is one of the most important physiological processes in insects.It is regulated by a serial of ecdysone cascade genes.Recently,lots o f microRNAs(miRNAs)were investigated in insects;however,their function in metamorphosis is largely unknown.In the present study,the dynamics of a small RNA population was investigated by RNA sequencing from the midgut of a lepidopteran pest Spodoptera litura during larval-pupal metamorphosis.A total of 101 miRNAs were identified,and 75 miRNAs were differentially expressed during the metamorphic process.The relationship between these differentially expressed miRNAs and 12 ecdysone cascade genes was analyzed by four classical software programs,and a multiple-to-multiple regulatory network was found to exist between these miRNAs and their targets.Among them,miR-14-3p and its two targets(EcR and E75)were chosen for further validation.MiR-14-3p had higher expression level in the 6 th instar larvae as compared with either the prepupae or pupae,which was opposite to that of both EcR and E75,two ecdysone cascade genes.Luciferase reporter assay confirmed that both EcR and E75 were regulated by miR-14-3p.Interestingly,the 3'untranslated regions are nearly identical to each other among different transcript variants of the ecdysone cascade genes,including EcR,USP,E75,E74,E78,E93,Hr3,Hr4,Hr39,K rhl and F tzfl.Thus,different transcript variants of one ecdysone cascade gene could be regulated by the same mi RNA.The above data suggest that the ecdysone signaling pathway is under the tight control of miRNA.These findings expand our understanding of the mechanism of insect metamorphosis and may also provide a novel possibility for the control of pest insects in the future.
基金the grants from Science and Technology Project of Guangzhou City (Grant No.201510010055)Natural Science Foundation of Guangdong Province (Grant No. 2015A030313390)National Natural Science Foundation of China (Grant No. 31672357).
文摘Silkworm mutants are valuable resources for both transgenic breeding and gene discovery. PiggyBac-based random insertional mutagenesis has been widely used in gene functional studies. In order to discover genes involved in silk synthesis, a piggyBac-based random insertional library was constructed using Bombyx mori, and the mutants with abnormal cocoon were particularly screened. By this means, a “thin cocoon” mutant was identified. This mutant revealed thinner cocoon shell and shorter posterior silk gland (PSG) compared with the wild type. The messenger RNA (mRNA) levels of all the three fibroin genes, including Fib-H, Fib-L and P25, were significantly down-regulated in the PSG of mutants. Four piggyBac insertion sites were identified in Aquaporin (AQP), Longitudinals lacking protein-like {Lola), Glutamyl aminopeptidase-like (GluAP) and Loc101744460. The mRNA levels of all the four genes were significantly altered in the silk gland of mutants. In particular, the mRNA amount of AQP, a gene responsible for the regulation of osmotic pressure, decreased dramatically immediately prior to the spinning stage in the anterior silk gland of mutants. The identification of the genes disrupted in the “thin cocoon” mutant in this study provided useful information for understanding silk production and transgenic breeding of silkworms in the future.
基金Cooperation Project in Industry,Education and Research of Guangdong Province and Ministry of Education of China(2009B090300349)a Cooperation Project of National Science Foundation of Guangdong Province(10351022401000000)
文摘Objective To establish a reverse-phase liquid chromatography method for the determination of seven alkaloids(magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine)in Fufang Zhenzhu Tiaozhi Capsule.Methods Chromatography was performed on a Dionex Acclaim C_(18)column(250 mm×4.6 mm,5.0μm)at 30℃.The mobile phase was composed of acetonitrile-potassium dihydrogen phosphate solution(0.015 mol/L,40:60,including 1.7 g/L sodium dodecyl sulfate and phosphoric acid used to regulate pH value to 3.0),with a flow rate of 1.0 mL/min.The detection wavelength was 270 nm.Results The calibration curves of magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine were linear in the range of 1.07-10.65,0.78-7.55,0.75-7.50,1.60-15.95,2.69-26.85,2.31-23.10,and 6.04-60.40 mg/mL.The average recoveries of magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine were 101.0%,101.2%,100.1%,100.0%,100.1%,101.1%,and 99.7%,respectively.Conclusion The method could be used for the quantitative determination of the preparation.
基金This study was supported by the Scientific Research Cultivation Fund for Young Faculty of South China Normal University （Grant No.： 14KJ15） and the National Natural Science Foundation of China （Grant No.： 31172265）.
文摘Juvenile hormone （JH） is one of the key insect hormones that regulate metamorphosis. Juvenile hormone diol kinase （JHDK） is an enzyme involved in JH metabolism and catalyzes JH diol to form a polar end product, JH diol phosphate that has no JH activity. In this study, a JHDK complementary DNA （cDNA） was cloned from Spodoptera litura and the structure and expression of the gene was characterized. The cDNA was 714 base pairs in length and encoded a protein of 183 amino acids with a molecular mass of 21 kDa and an isoelectric point of 4.55. Based on the structure, three putative calcium binding motifs and guanosine triphosphate-binding motifs were predicted in the protein. Modeling of the 3-D structure showed that the protein consisted of eight α-helixes linked with loops, with no β-sheets. The gene was expressed in the epidermis, fat body and midgut of fifth and sixth instar larvae. The expression level in the epidermis was lower than in the fat body and midgut. The gene was expressed at higher levels at the early stages than in the later stages of fifth and sixth instar midgut and fat body. The results suggest that this gene may be involved in the regulation of the JH titer in larvae ofS. litura.
