AIM: To investigate p16 gene alterations in gastric carcinoma. METHODS: 36 fresh tumor specimens taken from gastric cancer patients were analyzed for p16 gene deletion and mutation by polymerase chain reaction (PCR) and DNA sequencing. RESULTS: Homozygous deletion of exon 1 and exon 3 was observed in 2 cases and 3 cases of diffuse carcinoma, respectively. The frequency of homozygous deletion was 13.89%. No p16 gene point mutation was detected. CONCLUSION: The deletion of p16 genes may be related to gastric carcinogenesis.

With the development of medical research, the treatment of diabetes has not only limited to the internal medicine medications, surgery has become a new treatment. International Dibetes Federation officially recommended that metabolic surgery can be used for the treatment of obesity combined type 2 diabetes in 2011.

OBJECTIVETo establish a stable method of isolation, culture and cryopreservation of adult primary hepatocytes to provide potential hepatocyte resources for therapeutic usage in hepatocyte transplantation and bioartificial liver support systems for the treatment of acute and chronic liver diseases,and for experimental usage as an in vitro model of the liver.

METHODSAdult hepatocytes from 20 human donors undergoing partial hepatectomy were isolated using a two-step extracoporeal collagenase perfusion technique.Seven preincubation time points (2h,6h,12h,24h,36h,48h and 72h) were selected for optimization.After pre-incubation at 4 degrees C for 12-24h in HepatoZYME-SFM (the optimal condition),hepatocytes were microencapsulated using alginate-poly-L-lysine-alginate microcapsules,transferred to a complete medium containing 10% dimethyl sulphoxide and immediately placed into an isopropanol progressive freezing container for overnight freezing at -80 degrees C followed by immersion in liquid nitrogen the next day.During the post-thawing culture period,the cells were tested for albumin secretion,urea synthesis,cell cycling,transcription and protein synthesis (measuring mRNA and protein levels),and the morphological structure and pathology,for comparison with the features from before microencapsulated cryopreservation (PMC).

RESULTSThe viability and plating efficiency of the hepatocytes isolated using the two-step extracorporeal collagenase perfusion technique were 75.0+/-4.6% and 72.0+/-6.0%,respectively.The pre-incubation times of 12h and 24h (viability:61.4+/-4.8% and 62.0+/-5.6%; plating efficiency:3.2+/-5.8% and 62.6+/-3.6%,respectively) showed significantly higher albumin secretion than all other time points tested (F =40.3,all P less than 0.05).Compared with the immediate cryopreservation (immediately frozen control) hepatocytes,the PMC hepatocytes showed significantly better transcription and protein synthesis and higher albumin secretion and urea levels.The PMC group did not show a significantly different level of albumin production from the directly cultured hepatocytes (culture day 2:ll9.2ng/ml vs.131.36ng/ml,P =0.051; day 3:110ng/ml vs.120.4ng/ml,P=0.063; day 4:98.2ng/ml vs.109.8ng/ml,P more than 0.05).However,over culturing days 2,3 and 4,comparison of the PMC hepatocytes to the immediate cryopreservation hepatoeytes showed the former to have significantly higher secretion of albumin (119.2ng/ml vs.101.2ng/ml,110.0ng/ml vs.87.6ng/ml and 98.2ng/ml vs.73.8ng/ml; all P less than 0.05) and urea level (7.83 mug/ml vs.6.79 mug/ml,6.83 mug/ml vs.5.89 mug/ml and 5.85 mug/ml vs.4.83 mug/ml; all P less than 0.05).The post-thawed PMC hepatoeytes showed preservation of the morphological structure,while the immediate cryopreservation hepatocytes did not.

CONCLUSIONThe two-step extracorporeal collagenase perfusion technique after partial hepatectomy is a novel,simple,and reliable method for hepatocyte isolation.Pre-incubation at 4 degrees C for 12-24h before the microencapsulation cryopreservation allows for efficient recovery of functional and morphological integrity after thawing and provides viable hepatoeytes that may be useful for clinical applications in pharmacotoxicology,bioartificial liver therapy and cell therapy in humans.

Albumins ; Alginates ; Capsules ; Cell Cycle ; Cell Survival ; Cryopreservation ; Dimethyl Sulfoxide ; Hepatectomy ; Hepatocytes ; cytology ; Humans ; Perfusion ; Polylysine ; analogs & derivatives

Metabolic syndrome is a group of clinical syndromes characterized by overweight or obesity, hypertension, dyslipidemia, impaired glucose metabolism, and insulin resistance.It is a group of metabolic associated risk factors.Lumbar disc herniation (LDH) is one of the most common diseases nowadays, and its risk factors include obesity, diabetes, and abnormal blood lipid.In recent years, the relationship between LDH and metabolic syndrome (MetS) has been valued by scholars.Although some studies have suggested that MetS is related to LDH, the study of the correlation between MetS and LDH at home and abroad is relatively rare.To explore the possibility of using MetS for the prevention and control of LDH, this paper reviews the research progress of the relationship among MetS, the various components, and LDH.

Objective To investigate the clinical characteristics,diagnosis and treatment of congenital tracheobiliary fistula (CTBF) in children.Methods A case of CTBF admitted into the Department of Pediatric Respiration,Hubei Maternal and Child Health Care Hospital in 2016 was reported,and the related literatures were reviewed.The clinical features,diagnostic methods,treatment status and clinical outcomes of the disease were analyzed.Results The patient was 3 years and 7 months old.The main clinical manifestations were recurrent cough,pneumonia and atelectasis.CTBF was diagnosed by means of iodine oil radiography and treated with biological glue plugging under bronchoscopic guidance.So far,only 30 cases of CTBF have been reported in the English literatures,but only 2 cases in the Chinese literatures.The main clinical manifestations were cough,dyspnea,sputum or bile vomiting,aspiration pneumonia,atelectasis or emphysema.Diagnostic methods for CTBF included bronchoscopy,bronchial angiography and cholangiography,hepatobiliary scan,CT scan and magnetic resonance imaging.Except for this case treated with biological glue plugging under bronchoscopic guidance,all other patients were treated with surgery.The operation methods included fistula ligation,gastrostomy,liver resection,fistula jejunum Roux-en-Y anastomosis,hepatic hilum jejunum anastomosis,gallbladder jejunum anastomosis,etc.Only 4 cases died,and the rest of the patients recovered.Conclusions CTBF should be suspected in children with persistent chronic cough.Iodine oil radiography through bronchoscopy is a simple and feasible method for diagnosis of CTBF.Besides surgery,the lavage and the biological glue plugging method through bronchoscopy is also an effective way to treat CTBF in children without severe biliary malformation.

Objective:To investigate the association between quantified CT (QCT)-measured body composition and metabolic syndrome (MS) components in obese populations before bariatric surgery.Methods:A cross-sectional study. A retrospective analysis was conducted on a cohort of 97 obese patients scheduled for weight-loss surgery at the First Affiliated Hospital of Wannan Medical College from January 2021 to March 2023. The patients′ body mass index (BMI), biochemical parameters and body composition measurements obtained by QCT were recorded. The patients were stratified into groups based on gender, obesity severity and the number of MS components. Differences in body composition among the groups were compared. Additionally, the correlations between each body composition parameter and metabolic indicators were analyzed. The diagnostic efficacy of each body composition parameter for identifying obese individuals with different MS components was assessed using receiver operating characteristic (ROC) curve analysis.Results:There were 75 females (77.3%). Male obese patients had higher total abdominal fat area [(693.23±148.90) vs (574.99±114.89) cm 2, t=-3.958, P<0.001], visceral fat area [(289.65±57.67) vs (195.60±57.37) cm 2, t=-6.753, P<0.001], fat content of pancreatic head [27.45%(21.65%, 45.48%) vs 21.60%(17.6%, 26.9%), Z=-2.675, P=0.007], and skeletal muscle index [73.36(68.74, 81.26) vs 61.52(55.74, 66.41) cm 2/m 2, Z=-5.246, P<0.001]. With the increase of obesity, abdominal fat mainly increases in subcutaneous fat. With the increase of MS components (MS2 group, MS3 group, MS4 group, MS5 group), the abdominal fat area, abdominal fat/subcutaneous fat, liver fat content, pancreatic head fat content, and skeletal muscle index of patients all increased accordingly. In diagnosing the presence of two components of MS, area under the curve of visceral fat area was the largest (AUC=0.706, 95% CI=0.577-0.834). For diagnosing the presence of three, four and five components of MS, area under curve of liver fat content were all the largest (MS3=0.712, 95% CI=0.605-0.818; MS4=0.652, 95% CI=0.537-0.766; MS5=0.706, 95% CI=0.576-0.836). Conclusion:There are differences in QCT body composition among obese patients with different MS components, and there is a correlation between each body composition and MS component. Among them, intra-abdominal fat area and liver fat content are of great value in evaluating obese patients with different MS components.