Aim:To obtain recombinant human proinsulin C-peptide,a novel expression vector pEDCC was constructed to facilitate the expression and purification of C-peptide. Methods:Gene fragments encoding a truncated asparaginase fragment mutant,native C-peptide,a hinge fragment of human IgG1,an extra acid-labile dipeptide and a basic-amino-acid-riched octopeptide were introduced in turn into plasmid pET28a. The fusion protein ansB-C-hinge-DPKRKRKKSRNGSGR-C-peptide was expressed effectively as inclusion bodies after induced by lactose and partially purified by means of washing and ethanol fractionation. After being hydrolyzed,the polypeptide PKRKRKKSRNGSGR-C-peptide was liberated from the fusion partner. The N-terminal tetradecapeptide extension of C-peptide was subsequently cleaved by trypsin and removed by DE52 column. Results:The nucleotides sequence of plasmid pEDCC was confirmed to be identical with that of designed fusion protein. Recombinant human proinsulin C-peptide was obtained with high purity after purification. Conclusion:Employing truncated asparaginase as the fusion partner and basic-amino-acid-riched octopeptide to modulate isoelectric point is an effective approach to produce recombinant human proinsulin C-peptide.

Objective To determine the urinary polypeptide patterns of glomerulonephritis by magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technology. Methods Urinary samples of 29 healthy volunteers and 34 patients with glomerulonephritis, including 10 cases of IgA nephropathy (IgAN), 10 cases of membranous nephropathy (MN), 9 cases of minimal change disease (MCD) and 5 cases of lupus nephritis (LN), were collected and separated by magnetic bead,and were screened for polypeptide patterns with a novel high throughput method, MALDI-TOF MS. Results Under the relative molecular weights 10 000 Da, 85 protein peaks were detected in healthy controls group and 109 protein peaks were detected in glomerulonephritis group. Six peaks of 3371.5 Da, 4026.35 Da, 4085.32 Da, 4116.96 Da, 4126.32 Da and 9527.31 Da were up-regulated,while 8 peaks of 861.28 Da, 1205.41 Da, 1642.52 Da, 1913.15 Da, 1976.52 Da, 2087.74 Da, 2193.47 Da and 3015.57 Da were down-regulated by more than 2 folds (P<0.01) in glomerulonephritis group as compared to healthy controls. Urinary polypeptide patterns in different diseases differed significantly from each other, indicating specific disease pattern of polypeptide excretion. Conclusions MALDI-TOF MS is a fast, convenient and high throughput analyzing method capable of screening some relative specific, potential biomarkers from the urine of glomerulonephritis patients thus it possesses better clinical value.

Objective To evaluate the effect of early enteral nutrition (EN) on the pancreatic exocrine secretion in dogs with acute necrotizing panereatitis (ANP). Methods ANP model was induced by injection of mixtured solution of 5% sodium tanrecholate and trypsin into the pancreatic duct. Thirty dogs were randomly divided into total parenteral nutrition (TPN) group (n=5), duodenal PEPTI-2000VARIENT (DP) group (n=5), duodenal Nutrison MuhiFibre (DN) group (n=5), jejunal PEPTI-2000VARIENT (JP) group (n=5), jejunal Nutrison MuhiFibre (JN) group (n=5). The dogs were treated by either TPN or EN 24 hours after ANP model induction and the nutrition support lasted for 5 days. Serum amylase, LDH, lipase, secretin (SEC), cholecystokinin (CCK) and gastrin were measured at 1, 2, 3, 4, 5 d. Pancreatic juice was collected for 3 hours after TPN or EN started, and the amount of pancreatic juice and levels of proteinase, amylase, lipase, HCO3-, K+, Cl-, Na+ were determined. Dogs in each group were sacrificed at day 7. Histological and ultra-structure changes of the pancreatic tissues were evaluated pathologically. Results The levels of serum amylase, LDH, lipase, CCK, amount of pancreatic secretion and K+, Cl+, Na+ were not significantly different among these groups. The levels of plasma SEC and gastrin, HCO3-, proteinase, amylase, lipase in the duodenal nutrition groups were significantly higher than those in TPN group (P<0.05). The above mentioned parameters in the jejunal nutrition group were significantly lower than those in the duodenal group (P<0.05) and higher than those in the TPN group without significant difference. Among the 2 jejunal nutrition groups, the levels of plasma gastrin, HCO3- in pancreatic juice, proteinase, amylase, lipase in the JP group were significantly higher than those in the JN group (P<0.05). The above mentioned parameters in the DP group were significantly lower than those in the DN group (P<0.05). The amount of pancreatic secretion, HCO3-,K+, Cl+, Na+ were not significantly different among these groups. The pathological changes were similar among these groups, and the extent of pathological changes was relatively better in the JP group. The amount and density of intracytoplasm zymogen granules of pancreatic acinar cell were not significantly lower than those in the TPN group. Conclusions The delivery of nutrients to the proximal jejunum with elemental low-fat diets did not increase the pancreatic exacrine activity.

Objective To evaluate the efficacy and safety of wuling capsule compared with deanxit in treatment of patients with depression or anxiety status. Methods A total of 139 patients with depression or anxiety status were enrolled in a multi-center double blind clinical trial. All patients were randomized into two groups, 67 patients in wuling capsule group and 73 patients in deanxit group. Efficacy and adverse effects were evaluated with Hamilton Anxiety Scale (HAMA), Hamilton Depression Scale (HAMD),Pittsburgh Sleep Quality Index (PSQI) and Treatment Emergent Symptoms Scale (TESS) before treatment and 1, 2, 4, 6 weeks after treatment to verify the noninferiority of wuling capsule. Results There was no significant difference in score decline of HAMA and HAMD between the two groups (HAMA: wuling vs deanxit group: 9.0 ± 5.4 vs 10.3 ± 5.4, F = 2.11, P =0.148 ; HAMD: wuling vs deanxit: 9.1±5.6 vs 10.5±5.7, F = 1.61, P=0.207). There was no significant difference in response rate and remission rate between the two groups. No significant difference was found in score decline of PSQI between the two groups (wuling vs deanxit group: 4.1±4.5 vs 5.0 ± 4.2, F = 0.72, P=0.192). The incidence of adverse events was 41% in the wuling group and 55% in the deanxit group respectively. The main adverse effects were symptoms of gastrointestinal and neurology system. Conclusion Safe and well tolerated, wuling capsule improves the anxiety, depression and insomnia symptoms of patients with depression or anxiety status.

OBJECTIVETo determine the expression of Skp2 in different prostate cancer (PCa) cell lines and tissues, and explore its influence on the androgen receptor (AR) signaling pathway and development of castration-resistant prostate cancer (CRPC).

METHODSThe expression levels of Skp2 and AR in different PCa cell lines were detected by Western blot. After knockdown of Skp2 in the C4-2 and 22RV1 cells transfected with shRNA, the expressions of AR and P27 were determined and the activity of ARR3-Luc measured by dual-luciferase reporter gene assay following treatment with dihydrotestosterone (DHT). The expressions of AR and Skp2 in human naïve PCa or CRPC specimens were detected by immunohistochemical staining followed by analysis of their differences and correlation.

RESULTSThe Skp2 protein expression level was significantly higher in the C4-2 or 22RV1 cells than in the LNCaP cells. DHT treatment increased the expression of Skp2 in the C4-2 cells, but knock-down of Skp2 significantly up-regulated the expression of the well-known downstream protein P27 and down-regulated that of AR. Consistently, DHT treatment increased the activity of ARR3-Luc, while knockdown of Skp2 remarkably decreased it in the C4-2 and 22RV1 cells (P < 0.05). In addition, significantly higher expressions of Skp2 and AR were observed in the CRPC than in the naïve specimens (P < 0.05), with a positive correlation between the two proteins (r = 0.658 1, P < 0.05).

CONCLUSIONSkp2 can enhance the expression and transcription activity of the AR protein in CRPC cells or tissues and is promising to be a critical molecular therapeutic target.

Androgens ; pharmacology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Disease Progression ; Gene Knockdown Techniques ; Humans ; Male ; Neoplasm Proteins ; genetics ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; metabolism ; Receptors, Androgen ; genetics ; metabolism ; S-Phase Kinase-Associated Proteins ; physiology ; Transcriptional Activation ; Up-Regulation

Objective:To explore the efficiency of a new scoring system of gastric cancer screening for early gastric cancer in health examination population.Methods:The risk score of gastric cancer was assessed based on the new scoring system in health examination population. A notice for further gastroscopy was sent to the medium-risk and high-risk people. Gastroscopy was performed on those who agreed to undergo the examination.Results:From January to April 2019, a total of 5 357 people in health system visited the Physical Examination Center of Shanghai Songjiang Clinical Medical College of Nanjing Medical University for health examination. Seven hundred and forty people were classified as medium- and high-risk groups by the new screening system, 576 in medium-risk group, and 164 in high-risk group. Among them, 131 cases (17.70%) came for further gastroscopy, of whom 91 (69.47%) were in the medium-risk group and 40 (30.53%) in the high-risk group. After gastroscopy, 4 cases of gastric cancer and 1 case of esophageal cancer were detected, and both were early cancer. In the medium-risk group, 2 cases (2/91, 2.20%) of early gastric cancer and 1 case (1/91, 1.10%) of early esophageal cancer were found. In the high-risk group, 2 cases (2/40, 5.00%)of early gastric cancer were found. The tumor detection rate of high-risk group (5.00%) was higher than that of medium-risk group (3.30%), but there was no significant difference ( P>0.05). Conclusion:Risk stratification with the new scoring system of gastric cancer screening can improve the detection rate of early gastric cancer.

Docetaxel-based chemotherapy remains the first-line treatment for patients with metastatic castration-resistant prostate cancer (mCRPC) in China; however, the prognostic factors associated with effects in these patients are still controversial. In this study, we retrospectively reviewed the data from 71 eligible Chinese patients who received docetaxel chemotherapy from 2009 to 2016 in our hospital and experienced a reduction of prostate-specific antigen (PSA) level ≥50% during the treatment and investigated the potential role of time to nadir (TTN) of PSA. TTN was defined as the time from start of chemotherapy to the nadir of PSA level during the treatment. Multivariable Cox regression models and Kaplan-Meier analysis were used to predict overall survival (OS). In these patients, the median of TTN was 17 weeks. Patients with TTN ≥17 weeks had a longer response time to chemotherapy compared to TTN <17 weeks (42.83 vs 21.50 weeks, P < 0.001). The time to PSA progression in patients with TTN ≥17 weeks was 11.44 weeks compared to 5.63 weeks when TTN was <17 weeks. We found several factors to be associated with OS, including TTN (hazard ratio [HR]: 3.937, 95% confidence interval [CI]: 1.502-10.309, P = 0.005), PSA level at the diagnosis of cancer (HR: 4.337, 95% CI: 1.616-11.645, P = 0.004), duration of initial androgen deprivation therapy (HR: 2.982, 95% CI: 1.104-8.045, P = 0.031), neutrophil-to-lymphocyte ratio (HR: 3.963, 95% CI: 1.380-11.384, P = 0.011), and total PSA response (Class 1 [<0 response] compared to Class 2 [0-50% response], HR: 3.978, 95% CI: 1.278-12.387, P = 0.017). In conclusion, TTN of PSA remains an important prognostic marker in predicting therapeutic outcome in Chinese population who receive chemotherapy for mCRPC and have >50% PSA remission.

Objective To understand the situation of downward dislocation of intrauterine device (IUD) and the impact related to the effectiveness of IUD use, in China. Methods An epidemiological survey with cross-sectional, retrospective and prospective study designs was conducted to investigate 18 922 IUD users who were selected by a multi-phase stratified cluster sampling method. Results IUD's downward dislocation had been an important unsuccessful issue related to the IUD insertion that accounted for 20% of total the cases of failure. The top three failure outcomes would include extrusion,removal due to downward dislocation and unintended pregnancy. Respectively, the cumulative rates and the ranking due to IUD failure (per 100 women) in the first, third, sixth and twelfth month were shown as follows: extrusion appeared as 0.33%, 1.13%, 2.21% and 4.30%; removal as 0.10%, 0.37%, 0.80% and 2.34% ; while unintended pregnancy were 0.03%, 0.14%, 0.41% and 1.14%. Conclusion IUD' s downward dislocation made great impact on the effectiveness of IUD use that should call for attention from relative governmental sectors and researchers in the areas of prevention, diagnosis and treatment.

OBJECTIVE: To explore the genetic basis for pedigree affected with hereditary nephrogenic diabetes insipidus (HNDI). METHODS: Next generation sequencing (NGS) with an osteology system gene panel was carried out for the proband. Suspected mutation was validated by Sanger sequencing of two relatives with similar symptoms and two unaffected relatives from the pedigree. RESULTS: The proband was found to carry a c.856C>T mutation of the AVPR2 gene. The same mutation was detected in the two relatives with similar symptoms and one unaffected healthy relative. CONCLUSION The HNDI in this pedigree may be attributed to the c.856C>T mutation of the AVPR2 gene.
Diabetes Insipidus, Nephrogenic ; High-Throughput Nucleotide Sequencing ; Humans ; Mutation ; Pedigree ; Receptors, Vasopressin