Objective To evaluate the expression of KAII (CD82) gene inhibited by small interfering RNA (siRNA) in human pancreatic cancer cell line T3. Methods Four sequences of siRNA including A, B,C, D were designed, which were based on the KAI1 gene sequence using online RNA interfering designing software and lentivirus vector was built. Then they were used to transfect T3 cells by liposome 2000 and virus titer was determined. Empty vector containing siRNAd1 lentivrus particle ( MOI =5) was also used to infect T3 cells. The expression of CD82 mRNA was detected by real-time PCR. Results The expression of CD82 mRNA in normal control group, empty vector group, A group, B group, C group, D group were 1. 398 ±0.242,1. 311±0.048, 0. 664 + 0. 093, 0. 345 ± 0. 032, 0. 641 ± 0. 049 and 0. 147 ± 0. 049, respectively, the difference between the expression of CD82 mRNA in empty vector group and that of A, B, C, D groups was significant (P＜0.01 ). Conclusions RNAi was able to inhibit the expression of KAI1 gene CD82 in human pancreatic cancer cell line T3.

Objective To evaluate the effect of two polymorphisms(rs761737 and rs2269058) of RNF8 and the interactions with cigarette smoking and alcohol consumption on sperm DNA fragment index (DFI)and primary male infertility .Methods Based on case-control design ,polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology was used to de-tect the genotype of rs761737 and rs2269058 in RNF8 between 332 primary male infertile patients (composed by 87 patients of azoospermia ,166 patients of oligoasthenozoospermia and 79 patients of normozoospermia ) and 329 controls ,and Sperm Chromatin Dispersion(SCD) assay was used to assess sperm DNA fragment index (DFI) .Results Genotype and allele frequencies distribution of rs761737 and rs2269058 between cases and controls had no statistically significant difference (P>0 .05) .Sperm DFI in infertile group (46 .2 ± 22 .3)% was significantly higher than that of control group (21 .4 ± 9 .2)% (P<0 .05) ,stratified analysis suggested that Sperm DFI in oligoasthenozoospermia group (50 .0 ± 22 .1)% was also significantly higher than that of normozoospermia group (38 .2 ± 20 .7)% .The statistic differences of Sperm DFI in individuals who carried different genotypes of rs 761737 and rs2269058 in oligoasthenozoospermia group and normozoospermia group had no statistically significant difference (P>0 .05) .There was an inter-action between RNF8 rs2269058 and Cigarettes smoking(P<0 .05 ,OR=2 .37 ,95% CI 1 .06-5 .27) .Conclusion Although RNF8 rs761737 and rs2269058 have no effects on primary male infertility and sperm DFI ,cigarettes smoking increase the risk of primary male infertility in individuals who carry RNF8 rs2269058 AC+AA genotype .Sperm DFI is an important test to assess sperm quali-ty ,it is vital to reveal the etiology of primary male infertility and provide therapy guidance to clinicians .

This study is to explore the effects of quercetin (QUE) on the 3 week-old mice ovarian development and relative hormone levels. The 3 week-old mice were exposed to QUE (45, 25, and 5 mg x kg(-1) x hd(-1)) by gavage for 50 days. The estrous cycle during 50 days and the changes of hormone level such as FSH, LH, etc were monitored. Moreover, the ovaries were removed after sacrifice. The organ index was measured, and the ratios of different stages of follicles were analyzed by HE staining. Furthermore, the proportion of PCNA positive cells during all stages was detected by immunohistochemistry. The results showed that QUE could increase body weight of mice and reduce the anogenital distance (AGD) to some extent, and was able to disrupt mice's estrous cycle, but it could not extend or reduce the cycle regularity. It increased ovarian organ index with a dose-dependent manner. The proportion of the primordial follicle and secondary follicles rose obviously, and that of mature follicles', atretic follicles' and corpus luteums' reduced, while primordial follicle had no change. Immunohistochemistry analysis showed that QUE could effectively increase the percentage of proliferating cells in all kinds of follicles. Serum hormone assay showed that there were significant changes of FSH and LH levels. In summary, QUE showed an estrogen-like effect on mice's ovarian development. The weight of ovary, the proportion of all kinds of follicles, the development of ovarian cells and the level of plasma hormone in mice were altered obviously by oral administration of QUE.

Objective To investigate the effect of continuous veno-venous hemofiltration (CVVH) on pulmonary vascular permeability in the early stage of septic shock.Methods Fifty-one patients of septic shock admitted in intensive care unit of the First Affiliated Hopsital of Medical College,Zhejiang University between June 2010 and December 2011 were randomized into two groups by simple random method.Routine treatment was carried out to patients in group A,and routine treatment plus CVVH to patients in group B in the first 72 hours.Repeated respiratory mechanic hemodynamic measurements were done at baseline (Tb) before treatment,at 48 hours (T48) and 72 hours (T72) after the treatment.Pulmonary compliance (Cst),platform pressure (Pplat),oxygenation index (PaO2/FiO2),extravascular lung water index (EVLWI) and E-selectin determined at each time point were recorded in two groups.Results (1) Pplat were decreased significantly after treatment in two groups,and the Pplat in group B were all lower than those in group A at T48 and T72 (t =2.215,P ＜ 0.05 ; t =4.266,P ＜ 0.01).Cst were elevated after treatment in two groups,and Cst in group B were all higher than those in group A at T48 and T72 (t =2.516,P ＜0.05 ; t =3.052,P ＜ 0.01).(2) Compared to before treatment,PaO2/FiO2 increased significantly after treatment in two groups,and PaO2/FiO2 in group B were all higher than those in group A at T48 and T72 (t =2.732,P ＜0.01 ; t =3.511,P ＜0.01).(3) EVLWI were decreased significantly after treatment in two groups,and the EVLWI in group B were all lower than those in group A at T48 and T72 (t =2.597,P ＜0.05; t =2.125,P ＜ 0.05).ITBVI,CI,SVRI and MAP did not change over time in two groups compared with those at Tb (all P ＞ 0.05).(4) E-selectin were decreased significantly after treatment in two groups,and the E-selectin in group B were all lower than those in group A at T48 and T72 (t =2.154,P ＜0.05 ; t =3.581,P ＜ 0.01).Conclusion CVVH in early stage of septic shock can improve pulmonary vascular permeability and oxygenation,increase Cst,and decline EVLWI,with neither increased hemodynamics.

OBJECTIVE:To study the effects of Clebopride(CBP)bioadhensive sustained-release tablets on experimental gas-tric ulcer and gastrointestinal motility disorder. METHODS:Gastric ulcer rat model was induced by ethanol and aspirin,and then divided into model group (normal saline),common tablet (CBP tablet 0.072 mg/kg) and sustained-release tablet high-dose and low-dose groups (CBP bioadhensive sustained-release tablet 0.072,0.036 mg/kg);normal rats were included in normal control group (normal saline);they were given relevant medicine intragastrically,twice a day for sustained-release tablet,three times a day for other. Ulcer area were observed 2 and 4 days after medication to calculate healing rate of ulcer(n=6). Gastrointestinal mo-tility disorder mice model was induced by atropine,and then divided into model group (normal saline),common tablet group (CBP tablet 0.1 mg/kg)and sustained-release tablet high-dose,medium-dose and low-dose groups(CBP bioadhensive sustained-re-lease tablet 0.1,0.05,0.025 mg/kg);normal mice were included in normal control group(normal saline);they were given rele-vant medicine intragastrically,once a day,for consecutive 3 days. The rate of gastric emptying and small intestinal propulsion were detected (n=6). RESULTS:Compared with normal control group,ulcer area of rats increased in model group;compared with model group,that of rats decreased in common tablet group and sustained-release tablet high-dose,low-dose groups,with statisti-cal significance (P<0.01);healing rates of gastric ulcer were 32.35%-48.24% 2 days after medication,and those were above 70% 4 days after medication. Compared with normal control group,the rate of gastric emptying and small intestinal propulsion in mice decreased in model group;compared with model group,those of mice increased in common tablet group and sustained-re-lease tablet high-dose,medium-dose,low-dose groups. The effects of sustained-release tablet high-dose and medium-dose groups were better than that of common tablet group;those difference had statistical significance (P<0.01 or P<0.05). CONCLU-SIONS:CBP bioadhensive sustained-release tablets have im-provement effects against gastric ulcer of rats and gastrointesti-nal motility disorder of mice.

Objective To observe the inhibitory effect on metastasis and growth of pancreatic cancer in mice by injection of KAI1 gene in vivo. Methods Pancreatic cancer cell line MiaPaCa Ⅱ was used to construct the nude mice models bearing tumors, then the mice were divided into normal saline group, Ad group and Ad-KAI1 group. Since the 10th days of model construction, the Ad-KAI1 was injected every 7 d and repeated twice, then the tumor size, the weight of liver, lung and their pathologic changes were evaluated. Results The tumor sizes were not significantly different between the three groups. The weight of lung and liver of Ad-KAI1 group was (0.366±0.041) g and (1. 35±0.21) g, respectively; the weight of lung and liver of Ad group was (0.57±0.065) g and (1.58±1.828) g, respectively; the weight of lung and liver of control group was (0.66±0.13)g and (1.95±0.344)g, respectively. The difference between Ad-KAI1 group and control group was significantly different (t = 5.984, P < 0. 05), and there was no significant difference between Ad group and control group (t=1.089, P > 0.05). The number of pulmonary, liver and lymph node metastasis in Ad-KAI1 group was (1±1), (2±1) and (2±2), respectively; in Ad group was (6±2), (5 ±1), (10±2), respectively; in control group was (7±2), (6±2), (11±3), respectively. The difference between Ad-KAI1 group and control group was significantly different (t = 7.44, 4.34, 8. 16, P < 0.05), while the difference between Ad group and control group was not significantly different (t=0.92, 0.64, 0.42, P >0.05). Conclusions KAI1 gene directly injected into tumors of nude mice may inhibit the growth and metastasis of pancreatic cancer.

Objective:To investigate the clinicopathological features, treatment and prognosis of neuroendocrine carcinoma of the breast.Methods:Clinical data of 26 patients with neuroendocrine carcinoma of the breast admitted to the Northern Jiangsu People's Hospital from July 2013 to Mar 2021 were analyzed.Results:All 26 cases were female, the average aged of (62.81±11.95) years, the first clinical manifestations were painless breast masses, the average size being of (23.34±9.47) mm. At the time of diagnosis, regional lymph node metastasis was found in 4 cases, 1 case developed distant metastasis. Most patients' were on stage Ⅱ by TNM staging, molecular typing was Luminal A, and invasive mammary carcinoma with neuroendocrine differentiation was most common, with positive rates of ER and PR of 96%, the positive rate of CgA and Syn were 69% and 100%, and there was not positive expression of HER2. All patients received surgical treatment, 25 patients underwent postoperative adjuvant therapy. Twenty-five patients were followed up for a median follow-up time of 39.50 months. During the follow-up, 3 cases developed distant metastasis, 1 case died, the mean survival time was (40.81±26.90) months, there was ao satistically significant difference compared with invasive mammary carcinoma ( t=1.291, P=0.209). The mean disease free interval is (39.96±27.58) months. The overall survival and disease free survival at 1, 2 and 5 years are 100%, 100% and 87%, respectively. Conclusions:Neuroendocrine carcinoma of the breast occurs more frequently in elderly women, often with large tumor size, low rate of regional lymph node and distant metastasis, moderate histological grade, early clinical stage, and the molecular typing is mostly Luminal A.The overall prognosis is fair.

OBJECTIVE: To explore the genetic basis for a pedigree affected with Charcot-Marie-Tooth (CMT) disease through high-throughput sequencing. METHODS: Potential variants of the genes associated with CMT were screened by next-generation sequencing (NGS) of the members of the pedigree. RESULTS: NGS has revealed that the two affected sisters both harbored homozygous c.1A>G variant of the GDAP1 gene, which caused replacement of the first amino acid Methionine by Valine (p.Met1Val). Their parents were both carriers of the heterozygous c.1A>G variant. The variant was unreported previously and has an extremely low frequency in the population. Meanwhile, one of the sisters and the mother also carried heterozygous c.710A>T variant of the BAG3 gene. CONCLUSION The homozygous c.1A>G variant of the GDAP1 gene probably underlay the CMT in both children. Above result has enabled clinical diagnosis and genetic counseling for this pedigree.
Adaptor Proteins, Signal Transducing/genetics* ; Apoptosis Regulatory Proteins/genetics* ; Charcot-Marie-Tooth Disease/genetics* ; Child ; Female ; Fibula/abnormalities* ; Homozygote ; Humans ; Mutation ; Nerve Tissue Proteins/genetics* ; Pedigree

Objective:To investigate the clinicopathological differences in laterally spreading tumor (LST) from the rectum and colon.Methods:Clinicopathological records of 198 patients with LST (116 cases in rectum,82 cases in colon) from the Second Xiangya Hospital of Central South University between January 2012 and January 2017 were evaluated.Results:A total of 198 colorectal LST were included.According to the endoscopic classification,nodular mixed type (LST-GM),homogeneous type (LST-GH),flat elevated type(LST-FE) and pseudodepressed type (LST-PD) were 127(64.1％),13(6.6％),41(20.7％) and 17(8.6％),respectively.LST-GM was predominant in the rectum (71.7％),while LST-FE was predominant in the colon (78.0％),with significant difference (P＜0.01).The mean size of LST was (52.03±35.62) mm or (25.37±11.56) mm in the rectum or the colon,with significant difference between them (P＜0.01).High grade intraepithelial neoplasia frequency was higher in the rectum than that in the colon (31.0％ vs 18.3％),while the low grade intraepithelial neoplasia frequency was lower in the rectum than that in the colon (61.2％ vs 75.6％) (both P＜0.05).The mean size of LSTGM and LST-GH diameter were larger in the rectum than that in the colon,and the malignant potential of LST-GM was higher in the rectum than that in the colon.The percentage of high grade intraepithelial neoplasia + invasive carcinoma was 41.8％ and 22.2％,respectively (both P＜0.05).LST in colon was mostly treated with endoscopic mucosal resection,while LST in rectum was treated by endoscopic submucosal dissection predominantly.Conclu sion:LSTs from the rectum and colon show different clinicopathological characteristics to some extent.LST-GM is predominant in the rectum,while LST-FE is predominant in the colon.The malignant potential of LST-GM is higher in the rectum than that in the colon.

Objective To study the influence of clinical nutritional support on the effects of mechanical ventilation (MV), and to find the factors affecting the outcome of patients undergoing MV. Methods A case-control study was conducted. The clinical data of 235 patients undergoing MV admitted to intensive care unit (ICU) of Tongde Hospital of Zhejiang Province from January 2015 to June 2017 were retrospectively analyzed. The patients were divided into two groups according to whether weaning successfully within 7 days. The clinical data of patients in the two groups were collected including gender, age, acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score, underlying disease, nutritional indicators, nutritional support, and complications. The outcome of withdrawal within 7 days was served as a dependent variable, all observed indicators were served as independent variables, and Logistic regression analysis was carried out to screen the influencing factors of the weaning results within 7 days. Results 235 patients undergoing MV were enrolled, 128 patients were successfully withdrawn within 7 days, and 107 were unsuccessfully withdrawn. Compared with the successful weaning group, the patients of weaning failure group were older, and had higher APACHEⅡ score and lower albumin (Alb) and hemoglobin (Hb), more patients with internal medical underlying diseases and receiving parenteral nutrition (PN) and mixed nutrition, and the incidences of secondary infection, vomiting, abdominal distension, abnormal bowel sound, gastric retention, and diarrhea were higher. However, there was no statistical significance in gender between the two groups. The variables of statistical significance in univariate analysis were enrolled in the multifactor analysis model showing that age [odds ratio (OR) = 1.269, 95% confidence interval (95%CI) = 1.119-1.439, P < 0.001], APACHEⅡ score (OR = 1.643, 95%CI = 1.423-1.897, P < 0.001), internal medical underlying diseases (OR = 6.298, 95%CI = 4.012-9.887, P < 0.001), secondary infection (OR = 8.323, 95%CI = 2.568-26.975, P < 0.001), abdominal distension (OR = 3.368, 95%CI = 1.586-7.152, P = 0.002), abnormal bowel sounds (OR = 2.856, 95%CI = 1.215-6.713, P = 0.017), gastric retention (OR = 1.996, 95%CI = 1.183-3.368, P = 0.010), diarrhea (OR = 3.035, 95%CI = 1.337-6.890, P = 0.008) were risk factors for unsuccessful weaning,and compared with PN, enteral nutrition (EN; OR = 0.191, 95%CI = 0.098-0.372, P < 0.001) and mixed nutrition (OR = 0.375, 95%CI = 0.150-0.938, P = 0.037) were protective factors of successful weaning. The gender, Alb and Hb before and after MV, vomiting, gastrointestinal hemorrhage were not associated with weaning outcome within 7 days. Conclusions Elder, high APACHEⅡ score, internal medical underlying diseases, or secondary infection, abdominal distension, abnormal bowel sounds, gastric retention, diarrhea were risk factors of weaning failure within 7 days in patients undergoing MV. Compared with PN, EN and mixed nutrition were protective factors for successful weaning. For patients undergoing MV, EN should be performed early in the case of full recovery, hemodynamic stability, and serious metabolic disorders.