Effects of fermented cottonseed meal inclusions on growth performance, serum biochemical parameters and hepatic lipid metabolism of geese during 28-70 d of age.

The aim of this study was to investigate the effects of solid-state fermented cottonseed meal (FCSM) inclusion levels on the growth performance, serum biochemical parameters and hepatic lipid metabolism in geese from 28 to 70 d of age. A total of 288 twenty-eight-d-old male geese were randomly divided into 4 treatments with FCSM levels of 0, 5, 15 and 25% including 0, 22.74, 67.33, 111.27 mg FG/kg diet, respectively. Each treatment contained 6 replicates and 12 birds per replicate. Treatments of FCSM inclusions from 0 to 25% had no effect on growth rate and feed intake in geese during d 28 to 70. The F/G ratio was increased (P < 0.05) in geese fed the diet with 25% FCSM compared with birds fed the diet with 0% FCSM. Treatment with 25% FCSM levels had no effect on the contents of TC, TG, HDL-C, LDL-C, but increased (P < 0.05) AST and ALT activities in serum of geese at d 70. Treatment with 25% FCSM increased the contents of FG, HDL-C, TC, C18:2n6, C20:4n6 and PUFA and decreased (P < 0.05) the contents of NEFA, SFA, MUFA in liver compared with treatment of 0% FCSM inclusion. Additionally, treatment with 25% FCSM decreased (P < 0.05) the PPARα, AMPK, and LXR mRNA expression related to lipid deposition, and increased (P < 0.05) PPARγ and ACC mRNA expression related to lipolysis in liver compared with birds fed the diet with 0% FCSM. Overall, treatment with 0 to 15% FCSM (<=67.33 mg FG/kg diet) had no adverse effects on the growth performance and lipid metabolism of geese. However, treatment fed 25% FCSM (111.27 mg FG/kg diet) decreased feed efficiency and promoted hepatic lipid deposition associated with the alteration of related gene expression in geese at 28 to 70 d of age.

[Comparison of 1-year outcomes between Roux-en-Y gastric bypass and sleeve gastrectomy with Roux-en-Y duodenojejunal bypass in the patients of obesity with type 2 diabetes mellitus].

Objective: To compare the 1-year effects of Roux-en-Y gastric bypass (RYGB) and sleeve gastrectomy with Roux-en-Y duodenal bypass (SG+RYDJB) on weight loss, remission of diabetes, and postoperative complications in patients with obesity and type 2 diabetes. Methods: A single-center retrospective cohort study was conducted at the First Affiliated Hospital of Nanjing Medical University from January 2020 to December 2020. Sixty-four patients with type 2 diabetes and body mass index (BMI) of 27.5-40.0 kg/m2 were included in this study and divided into the RYGB group (n=34) and the SG+RYDJB group (n=30). In both procedures, the biliopancreatic branch was measured 100 cm distal to the Treitz ligament, and the food branch was measured 100 cm distal to the gastric or duodenojejunal anastomosis. Patients were followed up by telephone or WeChat, a free messaging and calling app at 1, 3, 6, and 12 months postoperatively to determine their weight loss and remission of diabetes. The primary outcomes were the weight loss and reduction in blood glucose concentrations at 1 year after surgery and postoperative complications. Other postoperative changes, including body weight, BMI, percentage of total weight loss (%TWL), percentage of excess weight loss (%EWL), glycated hemoglobin A1c (HbA1c), and fasting blood glucose at 1 year after surgery were also assessed. Results: There were no significant differences in baseline data between the two groups (all P>0.05). No conversion to open surgery or death occurred in either group. Operation time was longer in the SG+RYDJB than the RYGB group (137.8±22.1 minutes vs. 80.0±24.9 minutes, t=9.779, P<0.001) and the incidence of perioperative complications was higher in the SG+RYDJB than the RYGB group (20% [6/30] vs. 2.9% [1/34], χ2=4.761, P=0.029). However, the postoperative hospital stay was similar between the two groups [3.0 (3.0, 4.3) days vs. 3.0 (4.0, 6.0) days, U=641.500, P=0.071]. Perioperative complications comprised small gastric pouch anastomotic leakage in one patient in the RYGB group and leakage (three patients) and bleeding (two patients with gastrointestinal bleeding and one with trocar site bleeding) in the SG+RYDJB group. Long-term complications were as follows. The incidence of anemia was significantly higher in the RYGB than the SG+RYDJB group (26.5% [9/34] vs. 3.3% [1/30], χ2=6.472, P=0.011). However, there were no significant differences in incidences of postoperative reflux, dumping syndrome, alopecia, diarrhea, constipation or foul-smelling flatus between the two groups (all P>0.05). Compared with 1 year before surgery, the body weights and fasting plasma glucose concentrations of patients in the SG+RYDJB and RYGB group (72.4±10.6 kg vs. 98.5±14.2 kg, respectively; 68.2±10.0 kg vs. 91.9±14.8 kg, respectively), BMI (25.2±2.9 kg/m2 vs. 34.3±4.2 kg/m2, respectively; 24.3±2.4 kg/m2 vs. 32.7±3.7 kg/m2, respectively) (5.5±1.6 vs. 10.6±3.3, respectively; 5.8±2.1 vs. 9.0±3.4, respectively); HbA1c (5.7±0.8 vs. 9.7±1.2, respectively; 9.1±1.9 vs. 5.9±0.9, respectively) were significantly lower at 1 year after surgery (all P<0.05). However, the % TWL (26.5%±6.0% vs. 25.6%±4.4%, t=0.663, P=0.510) and % EWL (109.1%±38.2% vs. 109.4%±40.3%, t=-0.026, P=0.026), rate of complete remission of diabetes at 1 year (80.0% [24/30] vs. 82.4% [28/34], χ2=0.058, P=0.810] did not differ significantly between the two groups (all P>0.05). Conclusions: Although SG+RYDJB surgery compared with RYGB is more difficult to perform, it can achieve similar weight loss and remission of diabetes and is associated with a lower incidence of anemia because of the preservation of the pylorus.

[Clinical efficacy and prognostic risk factors of salvage liver transplantation, rehepatectomy, and local ablation in the treatment of postoperative recurrence of hepatocellular carcinoma].

Objective: To investigate and analyze the clinical efficacy of salvage liver transplantation (SLT), rehepatectomy (RH), local ablation (LA), and prognostic risk factors in patients with postoperative recurrence of hepatocellular carcinoma. Methods: Clinical data of 145 patients with recurrent liver cancer in the 900th Hospital of the Joint Logistics Support Force of the People's Liberation Army from January 2005 to June 2018 were retrospectively collected. SLT group, RH group, and LA group included 25, 44, and 76 cases, respectively. Follow-up and statistics were recorded on the overall survival rate, relapse-free survival rate, and complications of the three groups of patients at 1, 2, and 3 years after surgery. Univariate and multivariate COX analyses were used to analyze the prognostic risk factors in patients with recurrent HCC. Results: The overall survival rates of 1, 2, and 3 years following surgery in the SLT, RH, and LA groups were 100.0%, 84.0%, 72.0%, 95.5%, 77.3%, 65.9%, 90.8%, 76.3%, and 63.2%, respectively, when the recurrence of liver cancer met the Milan criteria. The overall survival rate did not differ statistically between SLT and RH (P = 0.303) or between RH and LA (P = 0.152). There were statistically significant differences in recurrence-free survival between SLT and RH or RH and LA (P = 0.046). There was no statistically significant difference in the incidence of complications between SLT and RH or RH and LA (P > 0.017). Age > 65 years was an independent risk factor affecting the overall survival rate in patients with recurrent HCC. Age > 65 years and recurrence time < 24 months were independent risk factors affecting the recurrence-free survival rate in patients with recurrent HCC. Conclusion: SLT is the best treatment option when the recurrence of HCC meets Milan's criteria. RH and LA are the appropriate treatment plans for recurrent HCC when the liver source is limited.

[Analysis of early severe postoperative complications and risk factors in 4255 patients who underwent bariatric and metabolic surgery in a single cente].

Objective: To analyze the incidence of early severe complications following bariatric and metabolic surgery and the experience of their diagnosis, treatment, and risk factors. Methods: In this retrospective observational study, the clinical data of 4255 patients who underwent bariatric and metabolic surgery between May 2010 and May 2022 in the Department of Bariatric and Metabolic Surgery of the First Affiliated Hospital of Nanjing Medical University were retrospectively collected. Among these patients, 1125 were male and 3130 were female. The mean age and body mass index (BMI) of the patients at the time of operation were 31.3±4.5 years and 36.5±6.4 kg/m2, respectively. Regarding surgical type, 2397 patients underwent sleeve gastrectomy (SG), 489 underwent Roux-en-Y gastric bypass (RYGB), 1028 underwent sleeve gastrectomy plus jejunojejunal bypass (SG+JJB), and 341 underwent single anastomosis duodenal switch (SADS). The inclusion criteria were patients (1) with a Clavien-Dindo grade of ≥III; (2) who were undergoing SG, RYGB, SG-JJB, or SADS; and (3) who had complete clinical data. The exclusion criteria were patients (1) undergoing revisional surgery and (2) other operations during the bariatric and metabolic surgery. The Clavien-Dindo classification was used to analyze the incidence of early severe postoperative complications and their prognosis. Early severe postoperative complications were defined as Clavien-Dindo ≥ III complications within 30 days after surgery. Meanwhile, multivariate logistic regression model was used to identify risk factors of the complications. Results: Summary of early severe complications following bariatric and metabolic surgery: (1) of the 4255 patients, 22 (12 male and 10 female) exhibited early severe complications (0.52%). The mean age and BMI of these patients were 41.1±9.9 years and 36.9±8.2 kg/m2, respectively. Preoperatively, 7 patients had hypertension, 10 had type 2 diabetes mellitus, 1 had respiratory failure, and 1 had heart failure. The severe complications included 9 patients (0.21%) with grade IIIa, 11 (0.26%) with grade IIIb, 1 (0.02%) with grade IVa, and 1 (0.02%) with grade V complications. The incidences of severe postoperative complications in the different surgical procedures were 0.17% for SG (4/2397), 0.61% for RYGB (3/489), 0.58% for SG+JJB (6/1028), and 2.64% for SADS (9/341). The common severe complications were leakage (0.28%, 12 patients), bleeding (0.14%, 6 patients), and obstruction (0.05%, 2 patients). (2) Management of complications: Grade IIIa complications (including eight patients with leakage and one with severe inflammation) were treated with antibiotics, nasogastric and nutritional tube placements, and CT-guided drainage. For grade IIIa complications, five patients with bleeding were treated with reoperation, and all the patients recovered; four patients with leakage were treated with reoperation, wherein three were converted to RYGB and one patient underwent resuturing of the leakage site; two patients with obstruction were treated with adhesiolysis. The patient with grade IVa complication (including respiratory failure complicated with acid aspiration) was treated in the ICU. For the grade V complication, bleeding in a patient with SG+JJB was treated with reoperation, which confirmed the bleeding of short gastric vessels. Unfortunately, the patient died. (3) Risk factor analysis of early severe complications: univariate analysis detected that sex, age, type 2 diabetes mellitus, operation time, and surgical type were associated with postoperative complications (P<0.05). However, multivariate analysis indicated that an age of ≥31.3 years (odds ratio [OR] = 5.423, 95% confidence interval [CI]: 1.004-29.278, P=0.049) and surgical type (SADS: OR = 19.758, 95%CI: 5.803-67.282, P<0.001; RYGB: OR = 9.752, 95%CI: 2.456-38.723, P=0.001; SG+JJB: OR = 5.706, 95%CI: 1.966- 16.559, P=0.001) were independent risk factors of early severe complications following bariatric and metabolic surgery. Conclusion: Bariatric and metabolic surgery is safe. Its common postoperative complications include leakage, bleeding, and obstruction, which require early detection, diagnosis, and treatment to improve treatment outcomes. Age and surgical type are independent risk factors of early severe complications following bariatric and metabolic surgery.

Survival analysis of 80 elderly patients with esophageal squamous cell carcinoma receiving definitive concurrent chemoradiotherapy with S-1.

PURPOSE: This single-center retrospective study aimed to observe survival and prognosis of elderly patients with esophageal cancer receiving radical radiotherapy combined with S1 chemotherapy, and to preliminarily explore whether hematologic indicators or inflammatory indicators before radiotherapy are prognostic factors. MATERIAL AND METHODS: We retrospectively collected data of 80 elderly patients with esophageal cancer who had received radical concurrent chemoradiotherapy from January 2015 to July 2018. The patients were older than 70 years. Radiation therapy was delivered with intensity-modulated irradiation therapy (IMRT) or volumetric modulated arc therapy (VMAT), while the chemotherapy regimen was S1 alone. Toxicities were evaluated in accordance with the criteria of the Radiation Therapy Oncology Group. Baseline hematologic basic nutritional indicators, such as hemoglobin (HGB), albumin (ALB), and prealbumin (PAB), along with inflammatory indicators, such as the ratio of neutrophils to lymphocytes (NLR), the ratio of platelets to lymphocytes (PLR), were collected to preliminarily analyze their relationship with progression and survival. RESULTS: The median follow-up time was 39 months (range 30-65 months). The median overall survival and progression-free survival times were 24 and 17 months, respectively. The 1-, 2-, and 3-year overall survival rates were 76.5%, 48.1%, and 31.3%, respectively. The 1-, 2-, and 3-year progression-free survival rates were 57.5%, 37.8%, and 29.4%, respectively. T stage, clinical staging, and lesion region were independent prognostic factors for OS. No significant associations with prognosis were observed either for baseline hematologic or for inflammatory indicators (all P>0.05). The rates of esophagitis (grade 2 and above) and hematologic toxicity were 60% and 45%, respectively. CONCLUSION: Oral S-1 combined with definitive concurrent radiotherapy for elderly patients with esophageal cancer has a significant survival benefit. The toxicities are well tolerated. It seems that prognosis does not correlate with baseline hematologic nutritional indicators and inflammatory indicators.

[Follow-up study of fetal cardiac birth defects after prenatal diagnosis and graded counseling].

Objective: To explore accurate prenatal diagnosis, full-coverage graded counseling and follow-up for the fetus with cardiac birth defects (CBD). Methods: CBD fetus diagnosed prenatal by echocardiography from January 2018 to December 2020 in Guangdong Provincial People's Hospital were enrolled. Fetal CBD was graded (Ⅰ-Ⅵ) according to prognosis and possible operation time after birth, and the classification criteria and common diseases included were proposed. After the prenatal grading counseling, the outcome of the fetus was followed-up. The induced labor rate, live birth rate, prenatal and postnatal ultrasound diagnosis coincidence rate and other indicators were calculated. The disease composition ratio, prognosis of fetus with different grades and the outcome of integrated treatment were analyzed. Results: The detection rate of fetal CBD was up to 16.2% (1 971/12 188), 30 cases of which were excluded. A total of 1 941 cases were included in this study, including 196 cases (10.1%) of gradeⅠ, 433 cases (22.3%) of gradeⅡ, 615 cases (31.7%) of grade Ⅲ, 261 cases (13.4%) of grade Ⅳ, 388 cases (20.0%) of gradeⅤ, 48 cases (2.5%) of grade Ⅵ. Grade Ⅱ and gradeⅢ (the operation time was within 1 year after birth) accounted for 54.0% (1 048/1 941). The distribution of some diseases in different grades had obvious proportion advantage, which was representative. Among 1 747 CBD fetus, 736 cases (induced labor rate 42.1%) chose to terminate pregnancy due to CBD. Of the 1 010 live births, 975 cases (96.5%) had the same prenatal and postnatal diagnosis, 3 cases were missed diagnosis and 32 cases were misdiagnosed. The diagnostic accuracy of live births with severe and complex congenital heart disease was 383 out of 389 (98.5%). A total of 258 cases have received surgery or intervention. The age at the time of surgery or intervention was different among grades（χ²=47.3，P<0.001）. With the improvement of prognosis from gradeⅠ to Ⅴ, the live birth rate increased and the induced labor rate decreased accordingly; the difference between grades was significant（χ²=623.6，P<0.001）. Conclusions: Prenatal diagnosis and graded counseling is important in the integrated model. Fetal CBD grading could refine post-natal treatment strategies, guide delivery decisions and become an evaluation standard.

Survival and complications of cytoreductive surgery and hyperthermic intraperitoneal chemotherapy in elderly patients: a systematic review and meta-analysis.

OBJECTIVE: Peritoneal cancer is an uncommon form of terminal malignancy with substantial morbidity and mortality. While both young and elderly population groups with peritoneal cancer are treated by joint cytoreductive surgery and hyperthermic intraperitoneal chemotherapy, studies suggest that they might have a differential prognostic outcome in terms of postoperative morbidity and mortality. To date, only one review has attempted to evaluate the comparative impact of postoperative complications and overall mortality in these age groups. However, a recent publication of several high-quality cohort trials needs an update of the existing consensus. To compare the impact of cytoreductive surgery and hyperthermic intraperitoneal chemotherapy on postoperative complications and overall mortality in younger and elderly population groups. MATERIALS AND METHODS: A systematic search of the academic literature was performed according to the PRISMA guidelines across five databases: Web of Science, EMBASE, CENTRAL, Scopus, and MEDLINE. A random-effect meta-analysis was conducted to evaluate the comparative outcomes between the impact of cytoreductive surgery and hyperthermic intraperitoneal chemotherapy on postoperative complications and overall mortality in younger and elderly population groups. RESULTS: From 963 studies, 16 eligible studies that evaluated the comparative outcomes of morbidity and mortality between 3067 young and 878 elderly patients undergoing cytoreductive surgery and hyperthermic intraperitoneal chemotherapy were included. A meta-analysis revealed higher risks of postoperative complications (Odds ratio: 1.18, 95% C.I: 0.90 to 1.55) and overall mortality (3.28, 1.93 to 5.5) for elderly patients as compared to the younger patients undergoing cytoreductive surgery and hyperthermic intraperitoneal chemotherapy. There were no differences in risks of the onset of anastomotic leakage (1.0, 0.47 to 2.14) and duration of hospital stay (Hedge's g: 0.02, -0.08 to 0.14) between elderly and younger patients undergoing cytoreductive surgery and hyperthermic intraperitoneal chemotherapy. CONCLUSIONS: The study provides updated evidence regarding poor postoperative morbidity and mortality outcomes in elderly patients as compared to younger patients undergoing cytoreductive surgery and hyperthermic intraperitoneal chemotherapy and may help clinicians to better stratify the risks associated with the conventional management of peritoneal carcinomatosis in elderly population groups.

Application of Automatic Nucleic Acid Extractor Combined with Vacuum Concentrator in Forensic Science.

ABSTRACT: Objective To explore the application value of automatic nucleic acid extractor combined with vacuum concentrator in forensic DNA extraction. Methods Gradient samples of human peripheral venous blood were collected at 40, 80, 120, 160, 200, 240, 280 and 320 fold dilution. The samples of each gradient were treated with no inhibitor, black oil, rust, fruit acid, tin foil and indigo, respectively. The automatic nucleic acid extractor was used for DNA purification and extraction of the above samples. The extracted DNA eluent （6 µL） was taken for amplification directly, and the rest was concentrated by vacuum concentrator. DNA was amplified and examined using the Investigator 26plex QS kit before and after concentration. Results Only gradient samples treated with fruit acid obtained complete STR typing results at 40 fold dilution. The other 5 methods obtained complete STR typing results at 40-160 fold dilution. The results of STR typing after DNA concentration showed that the average peak height and detection rates of gene loci both increased to a certain extent, but the effect was not obvious. Conclusion The automatic nucleic acid extractor has an efficient inhibitor removal ability and high extracting efficiency of DNA. The vacuum concentrator can concentrate DNA samples to a certain extent. Combining the automatic nucleic acid extractor with the vacuum concentrator can improve the examination success rate of forensic materials.

[Current status of registered drug and vaccine pediatric clinical trials in China].

Objective: To evaluate the current status of the registered pediatric drug or vaccine clinical trials in China for the purpose of providing a reference for the development of pediatric clinical trials in China. Methods: We collected the data about registered pediatric clinical trials that were conducted from September 6, 2013(Mandatory registration start date) to September 6, 2019 (Cut-off date) at Chinadrugtrials.org.cn platform. The survey items included trial name and number, drug classification, sponsor's information, current trial status, completion status, etc. The clinical trials were categorized by drug group (includes chemical medicine, traditional Chinese medicine and natural medicine, biological products) and by vaccine group. Results: During the six years 349 pediatric clinical trials were registered on the platform, including 162 pediatric drug trials and 187 vaccine trials. The numbers of chemical drugs and biological products registered in 2018 were 23 and 11, respectively, the highest in the history. The number of pediatric clinical trials of traditional Chinese medicine and natural medicine was 11 in 2014, but from 2015 to 2018 only 2 to 4 trials were registered each year. The overall completion rates of the registered drug and vaccine clinical trials were 22.8% (37/162) and 41.7%(78/187), respectively. Only 42 international multicenter pediatric clinical trial projects were registered on the platform. The numbers of drug and vaccine phase Ⅰ clinical trials were 4 and 46, respectively. Thirty-six pediatric endocrine system agent clinical trials were carried out, with the largest number of all the drug categories registered on the platform. Conclusions: In recent years the number of registered pediatric drug and vaccine clinical trials increased in China. However, the number is still very limited. It is urgent to further promote the development of pediatric clinical trials.

Mineral requirements in ducks: an update.

Mineral nutrition plays a critical role in growth and bone mineralization in meat ducks as well as reproductive performance in duck layers and duck breeders. In addition to improving production performance parameters, minerals are also essential to support several enzymatic systems to enhancing antioxidant ability and immune function. This review explores the biological function and metabolism of minerals in the body, as well as mineral feeding strategy of various species of ducks. Topics range from mineral requirement to the physiological role of macroelements such as calcium and phosphorus and microelements such as zinc and selenium, etc. As with the improvement of genetic evolution and upgrade of rearing system in duck production, mineral requirements and electrolyte balance are urgent to be re-evaluated using sensitive biomarkers for the modern duck breed characterized by the rapid growth rate and inadequate bone development and mineralization. For duck breeders, mineral nutrition is not only required for maximal egg production performance but also for maintaining normal embryonic development and offspring's performance. Therefore, the proper amounts of bioavailable minerals need to be supplemented to maintain the mineral nutritional state of duck species during all phases of life. In addition, more positive effects of high doses microelements supplementations have been revealed for modern meat ducks subjected to various stresses in commercial production. The nutritional factors of mineral sources, supplemental enzymes, and antinutritional factors from unconventional ingredients should be emphasized to improve the effectiveness of mineral nutrition in duck feed formulation. Organic mineral sources and phytase enzymes have been adopted to reduce the antagonistic action between mineral and antinutritional factors. Therefore, special and accurate database of mineral requirements should be established for special genotypes of ducks under different rearing conditions, including rearing factors, environmental stresses and diets supplemented with organic sources, phytase and VD3.

Overexpression of miR-32 inhibits the proliferation and metastasis of ovarian cancer cells by targeting BTLA.

OBJECTIVE: The purpose of this study was to explore the role of microRNA-32 (miR-32) in ovarian cancer and the possible underlying mechanism. PATIENTS AND METHODS: Ovarian cancer tissues were collected from 100 patients diagnosed with ovarian cancer in our hospital. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) was used to detect the expression levels of miR-32 and its target gene in ovarian tissues. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay was conducted to detect the proliferation of ovarian cancer cells. Meanwhile, the transwell and wound healing assays were used to evaluate the migration and invasion abilities of ovarian cancer cells, respectively. Bioinformatics (including TargetScan, miRDB, and microRNA) were used to predict the target genes of miR-32. Furthermore, The Dual-Luciferase reporter gene assay was performed to verify the binding relationship. RESULTS: MiR-32 was significantly downregulated in both ovarian cancer tissues and cells. The overexpression of miR-32 significantly inhibited the proliferation, migration, and invasion of ovarian cancer cells. B and T lymphocyte associated (BTLA) was screened out as a target gene of miR-32. Furthermore, BTLA could counteract the effects of miR-32 on ovarian cancer cells. CONCLUSIONS: Acting as a suppressor gene, miR-32 inhibited the malignant behaviors of ovarian cancer cells by regulating its target gene BTLA.

LncRNA HOTAIR inhibited osteogenic differentiation of BMSCs by regulating Wnt/ß-catenin pathway.

OBJECTIVE: This study aims to investigate whether HOX transcript antisense RNA (HOTAIR) can participate in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by regulating the Wnt/ß-catenin pathway, thereby participating in the pathogenesis of osteoporosis. PATIENTS AND METHODS: We detected the expression level of HOTAIR in 60 osteoporosis patients and 60 normal controls by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). Meanwhile, BMSCs derived from human or rats were subjected to determination of HOTAIR level. Subsequently, the effects of HOTAIR on osteogenic differentiation were evaluated by the activity of Alkaline Phosphatase (ALP), Alizarin Red S (ARS) staining, ALP staining and osteogenic-specific gene expression. The expression level of proteins related to the Wnt/ß-catenin was determined by Western blot, and ALP activity was detected by ALP activity determination kit and alizarin red staining after knockdown or overexpression of HOTAIR, as well as the treatment of DKK1 or the Wnt pathway antagonist. Finally, osteoporosis model in rats was established by ovariectomy (OVX). We examined protein levels of HOTAIR, ß-catenin, CyclinD, C-myc, and Runx2 in rat bone tissues. Bone morphology was observed in each group as well. RESULTS: The serum and BMSCs levels of HOTAIR in patients with osteoporosis were remarkably higher than that in normal people. Inhibition of HOTAIR induced increased ALP activity increased osteogenic marker genes and enhanced number of calcified nodules in BMSCs. However, the overexpression of HOTAIR exhibited the opposite effects. HOTAIR inhibited the expression level of Wnt/ß-catenin pathway-related protein. Also, Wnt pathway antagonist DKK1 partially reversed the regulatory effects of HOTAIR on Wnt/ß-catenin. DKK1 treatment markedly reduced the promotive effect of HOTAIR knockdown on ALP activity, ALP content and calcification ability of BMSCs. DKK1 administration in rats undergoing OVX showed worse bone morphology relative to controls. Protein levels of HOTAIR, ß-catenin, CyclinD, C-myc and Runx2 remarkably downregulated in OVX rats administrated with DKK1. CONCLUSIONS: HOTAIR inhibits osteoblast differentiation of rat BMSCs. The underlying mechanism of which may be related to the mediation of Wnt/ß-catenin pathway.

[Etiology analysis and vestibular assessment of bilateral vestibular vestibulopathy].

Objective:To define clinical and laboratory characteristics of bilateral vestibulopathy(BVP) and to propose diagnostic criteria of this disorder based on clinical and laboratory vestibular function test findings.Method:Forty-two case series with a clinical suspicion of BVP were retrospectively analyzed, in an attempt to determine etiology. Presenting auditory-vestibular symptoms, bedside dynamic visual acuity tests and laboratory test were reviewed, including bithermal caloric test, rotatory chair tests, video head impulse test (vHIT), vestibular-evoked myogenic potentials (VEMP).Result:Among these 42 patients, dizziness was seen in 42 cases(100%), oscillopsia was seen in 21 cases(50%), hearing loss was seen in 30(71.4%). Eight cases(19%) had tinnitus. Twenty-five cases showed vestibular loss in dynamic visual acuity test (69.4%). Definite diagnosis of complete BVP was made in 36 patients when the patients showed abnormal findings on caloric test, rotatory chair test and vHIT in addition to the symptoms. Whereas probable diagnosis of partial BVP was obtained in 6 patients with abnormal caloric test and rotatory chair test but no pathological vHIT. VEMP (ocular or cervical) could be recorded in 20 patients. Fourteen cases were caused by ototoxic drugs while no causes could be determined in 6 cases among these 42 cases.Conclusion:The diagnosis of BVP is a challenge. Vestibular laboratory test battery which reflect full frequency function of VOR has great value to confirming the diagnosis and differentiate complete BVP to partial BVP. Diagnosis standard shall be made combining clinical history, characteristic symptoms and the results of auditory-vestibular function testing. Ototoxic drugs contribute most considering etiology.

[Effect of endoplasmic reticulum stress in trophocytes on the pathogenesis of intrahepatic cholestasis of pregnancy].

Objective: To evaluate the effect of endoplasmic reticulum stress in trophocytes, in patients with intrahepatic cholestasis of pregnancy (ICP). Methods: Sixty-one pregnant women who were hospitalized in Women's Hospital, School of Medicine, Zhejiang University from January to December 2015 were recruited. Thirty-one women who were diagnosed as ICP were defined as the ICP group and 30 healthy pregnant women were defined as the control group. The localization and expression intensity of glucose regulated protein 78 (GRP-78) in placental tissues were detected by immunohistochemistry technique. Electronic microscope was used to observe ultra-microstructure change of the endoplasmic reticulum in trophocytes and cell line Swan71. Reverse transcription (RT)-PCR and western blot were used to investigate the expression of GRP-78 mRNA and protein in Swan 71 cell. Results: (1) GRP-78 protein was mainly expressed in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts. The protein expression of GRP-78 in placentas of the ICP group (13.2±2.4) was significantly higher than that in the control group (7.8±1.3, P<0.01). (2) The volume of endoplasmie reticulum did not increase and the microvilli developed well, with no swelling and no expansion of endoplasmic reticulum in the control group.In the ICP group, microvilli injury, endoplasmic reticulum edema were found; the volume of endoplasmic reticulum increased, with dilation, vacuolation and significant degranulation. After treated with 100 µmol/L cholyglycine for 24 hours, universal dilatation of the endoplasmic reticulum were seen in the Swan71 cells. (3) In Swan71 cells, cholylglycine displayed a concentration-dependent up-regulation on the expression of GRP-78. The expressions of GRP-78 mRNA in 0, 25, 50, 100 µmol/L cholylglycine experimental group were 1.01±0.17, 2.17±0.16, 5.47±0.36, 5.65±0.82, respectively. The expression of GRP-78 protein in 0, 25, 50, 100 µmol/L cholylglycine experimental group were 1.01±0.04, 1.17±0.15, 1.33±0.13, 1.73±0.13, respectively. The expression of GRP-78 mRNA and protein in 100 and 50 µmol/L cholylglycine experimental group were significantly higher than 0 µmol/L (all P<0.01). Conclusion: The obvious expansion of endoplasmic reticulum and the increased expression of GRP-78 in trophocytes indicated that endoplasmic reticulum stress of trophocytes may be involved in the pathogenesis of ICP.

The reliability and validity of the Unified Classification System of periprosthetic femoral fractures after hip arthroplasty.

The Unified Classification System has expanded and updated the Vancouver Classification System and applied treatment principles to all periprosthetic fractures. This study assessed the reliability and validity of the Unified Classification System for femur fractures after hip arthroplasty. Thirty-eight radiographs were finally evaluated by 6 observers, 3 consultants and 3 trainees. Each observer read the radiographs on 2 separate occasions, independently, at least 2 weeks apart. Interobserver and intraobserver agreement and validity were analyzed, using weighted κ statistics. The mean κ value for interobserver agreement was found to be 0.849 (0.838-0.871) for consultants (almost perfect agreement) and 0.761 (0.707-0.836) for the trainees (substantial agreement). Intraobserver κ values ranged from 0.740 to 0.903, showing substantial to almost perfect agreement. Validity analysis of 23 type B cases revealed 79.710% agreement within B1, B2, and B3 subgroups with a mean κ value of 0.694 (0.670-0.741) (substantial agreement).

Circulating DNA for detection of gastric cancer.

OBJECTIVE: Gastric cancer (GC) is one of the most common malignant tumors worldwide, particularly, prevalent in China. Despite the decreasing incidence of GC in China, the 5-year survival rate is still not over 30% yet. Therefore, early diagnosis and therapeutic outcome evaluation of GC remains as the issue to be resolved in a clinical setting. MATERIALS AND METHODS: Recent studies have found the presence of a certain amount of circulating DNA in the peripheral blood of patients with malignant tumor and shown that these free DNA bear tumor-specific genetic information. The circulating DNA detection includes quantitative and qualitative methods and analysis. Combined monitoring of changes in circulating DNA levels and aberrant alteration of relevant tumor genes is likely to provide comprehensive real-time information to patients. RESULTS: Under normal conditions, oncogene presents in the form of proto-oncogene such as K-ras, which is in non-carcinogenic status under the influence of tumor suppressor gene. When tumor suppressor gene is damaged or mutated of oncogene itself is induced for instance P53, oncogene is then activated and induces tumorigenesis. However, compared to gene mutation detection, the detection of DNA methylation is relatively more well-developed and stable. CONCLUSIONS: This article reviews the current status of the research on circulating DNA in the diagnosis, assessment of response to therapy and prognostic evaluation in GC. In addition, the advantage, current issue and prospect of using circulating DNA as tumor marker are also analyzed.

Genome-wide association study on antipsychotic-induced weight gain in the CATIE sample.

Antipsychotic-induced weight gain (AIWG) is a common side effect with a high genetic contribution. We reanalyzed genome-wide association study (GWAS) data from the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) selecting a refined subset of patients most suitable for AIWG studies. The final GWAS was conducted in N=189 individuals. The top polymorphisms were analyzed in a second cohort of N=86 patients. None of the single-nucleotide polymorphisms was significant at the genome-wide threshold of 5x10(-8). We observed interesting trends for rs9346455 (P=6.49x10(-6)) upstream of OGFRL1, the intergenic variants rs7336345 (P=1.31 × 10(-5)) and rs1012650 (P=1.47 × 10(-5)), and rs1059778 (P=1.49x10(-5)) in IBA57. In the second cohort, rs9346455 showed significant association with AIWG (P=0.005). The combined meta-analysis P-value for rs9346455 was 1.09 × 10(-7). Our reanalysis of the CATIE GWAS data revealed interesting new variants associated with AIWG. As the functional relevance of these polymorphisms is yet to be determined, further studies are needed.The Pharmacogenomics Journal advance online publication, 1 September 2015; doi:10.1038/tpj.2015.59.

Compound heterozygous mutations in TTC7A cause familial multiple intestinal atresias and severe combined immunodeficiency.

Familial multiple intestinal atresias is an autosomal recessive disease with or without combined immunodeficiency. In the last year, several reports have described mutations in the gene TTC7A as causal to the disease in different populations. However, exact correlation between different genotypes and various phenotypes are not clear. In this study, we report identification of novel compound heterozygous mutations in TTC7A gene in a Malay girl with familial multiple intestinal atresias and severe combined immunodeficiency (MIA-SCID) by whole exome sequencing. We found two mutations in TTC7A: one that destroyed a putative splicing acceptor at the junction of intron 17/exon 18 and one that introduced a stop codon that would truncate the last two amino acids of the encoded protein. Reviewing the recent reports on TTC7A mutations reveals correlation between the position and nature of the mutations with patient survival and clinical manifestations. Examination of public databases also suggests carrier status for healthy individuals, making a case for population screening on this gene, especially in populations with suspected frequent founder mutations.

Effect of RNAi-mediated silencing of Livin gene on biological properties of colon cancer cell line LoVo.

This study aimed to investigate the effect of RNAi-mediated silencing of the Livin gene on biological properties of the colon cancer cell line LoVo. Interference vectors pSilencer4.1-Ll and pSilencer4.1-L2 targeting the Livin gene were constructed and transfected into LoVo cells. The expression of the Livin gene was determined by RT-PCR and Western blotting. The apoptosis, cell cycle, colony formation, proliferation of LoVo cells, as well as their sensitivity to cisplatin, were detected by flow cytometry, colony formation assay and MTT. Livin mRNA and protein expression in LoVo cells could be effectively silenced by pSilencer4.1-Ll but not pSilencer4.1-L2. In the pSilencer4.1-Ll transfection group, the apoptosis rate of LoVo cells was significantly higher than in the control group (24.2 ± 3.2 vs 8.1 ± 1.4%, P < 0.01), and after 72 h, cell proliferation was clearly decreased (about 70% inhibition). Compared with the control group, the colony formation rate in pSilencer4.1-Ll transfection group was obviously decreased (15 ± 4.6 vs 85 ± 5.8%, P < 0.01), with increased proportion of S phase cells (45.7 ± 4.9 vs 28.0 ± 3.0%, P < 0.01), decreased proportion of G1 phase cells (43.0 ± 5.2 vs 62.8 ± 5.1%, P < 0.01), and increased sensitivity to cisplatin (apoptosis rate increased from 43.4 ± 6.9 to 65.3 ± 6.2%, P < 0.01). pSilencer4.1-Ll can effectively silence Livin gene expression in LoVo colon cancer cells, inhibit cell proliferation and colony formation, induce apoptosis, and enhance sensitivity to cisplatin.

Cadmium toxicity is alleviated by AtLCD and AtDCD in Escherichia coli.

AIMS: Arabidopsis thaliana l- and d-cysteine desulfhydrases (AtLCD and AtDCD) are two important H(2) S-generating enzymes. This study determined the effects of H(2) S derived from AtLCD and AtDCD on cadmium (Cd) toxicity in Escherichia coli. METHODS AND RESULTS: AtLCD and AtDCD were cloned into pET28a vectors and transformed into wild-type E. coli strain BL21(DE3), named BL21(LCD) and BL21(DCD). In the induced BL21(LCD) and BL21(DCD) compared with wild type, significantly higher H(2) S generation rates were observed. Additionally, higher survival rates, reduced contents of malondialdehyde (MDA) and hydrogen peroxide (H(2) O(2)), decreased activities of superoxide dismutase and catalase under 220 µmol l(-1) Cd stress were noted. We obtained similar results in the wild type treated with NaHS, a H(2) S donor. The above changes were substantially counteracted by the mixture of ammonia and pyruvic acid potassium (NH(3) + C(3) H(3) KO(3)), a synthetic inhibitor of H(2) S. CONCLUSIONS: AtLCD and AtDCD catalyse the H(2) S production, generating an ameliorating effect against Cd-induced oxidative stress and resulting in E. coli resistance to Cd toxicity. SIGNIFICANCE AND IMPACT OF THE STUDY: H(2) S as a gasotransmitter is certified to have an ameliorating effect against Cd toxicity, thus providing information for further research regarding the role of H(2) S in regulating resistance to the heavy metal stress in organisms.