OBJECTIVE: To establish an HPLC method for the content determination of prostaglandin A1 and prostaglandin B1 in Alprostadil for injection.METHODS: The determination was performed on Alltech Alltima C18 column with mobile phase consisted of phosphate puffer(pH=6.3)-acetonitrile-methanol(70 ∶ 25 ∶ 5) at a flow rate of 1.5 mL? min-1.The detection wavelength was set at 196 nm.The column temperature was set at room temperature and the injection volume was 20 ?L.RESULTS: The prostaglandin A1 and prostaglandin B1 were well separated from main component and other impurities.The linear range of prostaglandin A1 and prostaglandin B1 were 0.175～19.00 ?g?mL-1(r=0.999 7) and 0.23～19.90 ?g?mL-1(r=0.999 2).The contents of prostaglandin A1 in 3 batches of samples were 4.7%,4.9% and 4.3%,and the contents of prostaglandin B1 in 3 batches of samples were 0.6%,0.8% and 0.5% respectively.CONCLUSIONS: This method is proved to be simple,specific and suitable for the content determination of related substances in Alprostadil for injection.

Objective To evaluate the clinical features and guanosine triphosphate cyclohydrolase 1 (GCH-1) gene mutation in a family with dopa-responsive dystonia (DRD).Methods The clinical features of this family were collected and their peripheral blood samples were screened for mutation in GCH-1 gene using PCR and DNA direct sequencing.Results The clinical features among each patient in this family were different.But all affected family members had quite a good response to levodopa treatment without significant adverse reactions.DNA test showed an AT deletion mutation at point of 631-632 in the 6th exon of GCH-1 gene in 5 affected members and 1 asymptomatic immediate family member.Conclusions Clinical heterogeneity is an important characteristic of DRD and clinical symptoms vary intra-families.Same gene type may cause different phenotype and not all carriers are patients.The deletion mutation at point of 631-632 in the 6th exon of GCH-1 gene should be considered as a pathogenic mutation for DRD.

BACKGROUND: Previous studies have demonstrated that LRP16 is an estrogen-responsive gene. Its expression level is strongly associated with the proliferation and invasive growth of human breast cancer cells.OBJECTIVE: To construct a LRP16 targeting vector and screen mouse embryonic stem cell clones with homolougous recombination of an inactive LRP16 gene.DESIGN: Constructing an inserting inactivation target by inserting SA-RIES-β geo expression cassette.SETTING: Bioregulatory Laboratory of the Third Medical Department of Kyushu University in Japan and Department of Molecular Biology, General Hospital of Chinese PLA.MATERIALS: The materials used here were mainly provided by the Bioregulatory Laboratory, the Third Medical Department of Kyushu University in Japan. The mouse genomic library in pBeloBAC11 Vector was purchased from lnvitrogen Corp. The competent TopF10 was purchased from Beijing Tiangen Biotech Corp. pcDNA3.1(+) vector was kept in our laboratory. Mouse ES cells were provided by Kyushu University.METHODS: The experiment was performed in Kyushu University and Department of Molecular Biology of PLA General Hospital from November 2004 to May 2005. Targeting sequence of LRP16 gene was obtained from 129 mouse genomic Bacterial Artificial Chromosomes library based on polymerase chain reaction (PCR) screening. The SA-RIES-β geo fragment was inserted within LRP16 fifth exon to inactivate LRP16. ES cells were screened with G418 and the homologously recombinant clone was identified by Southern blot analysis.MAIN OUTCOME MEASURES: Clones with homologous recombination.RESULTS: The LRP16 fragment including exon 5 to 11 was subcloned into the pBluescript SK Ⅱvector. Restriction map demonstrated that the SA-IRES-β geo fragment was correctly inserted into the LRP16 fifth exon. Southern blot results showed that there was an ES clone with targeting sequence homologously inserted.CONCLUSION: A LRP16 gene targeting vector is constructed and a homologous recombinant is obtained.

Objective To investigate the predictive value of cumulative body mass index (cumBMI) on new-onset cholelithiasis.Methods The retrospective cohort study was conducted.The data of 31 794 subjects who participated health examination at the Kailuan Hospital,Kailuan Linxi Hospital,Kailuan Zhaogezhuang Hospital,Kailuan Tangjiazhuang Hospital,Kailuan Fan'gezhuang Hospital,Kailuan Lyujiatuo Hospital,Kailuan Jinggezhuang Hospital,Kailuan Linnancang Hospital,Kailuan Qianjiaying Hospital,Kailuan Majiagou Hospital and Kailuan Branch Hospital in 2006,2008,2010,2012 and 2014 were collected.All the subjects were allocated into 4 groups according to squartiles of cumBMI:7 949 with cumBMI＜ 140.81 kg/m2 ×year in the Q1 group,7 946 with 140.81 kg/m2×year≤ cumBMI＜ 159.69 kg/m2 ×year in the Q2 group,7 949 with 159.69 kg/m2×year≤cumBMI＜ 180.49 kg/m2 ×year in the Q3 group and 7 950 with cumBMI ≥ 180.49 kg/m2×year in the Q4 group.All the subjects received respectively the five health examinations in 2006,2008,2010,2012 and 2014 at the same place.Epidemiological investigation,anthropometric parameters and biochemical indicators were collected.Observation indicators:(1) incidence of cholelithiasis in the 4 groups;(2) risk factors analysis affecting newonset cholelithiasis:sex,age,cumBMl,BMI,drinking,smoking,physical exercise,hypertension,diabetes,C-reactive protein (CRP),triglyceride (TG) and total cholesterol (TC).Measurement data with normal distribution were represented as-x±s and comparisons among groups were analyzed using the one-way ANOVA.Pairwise comparison and homogeneity of variance were done using the LSD test.Heterogeneity of variance was done using the Dunnett's T3 test.Measurement data with skewed distribution were described as M (Q) and comparisons among groups were analyzed using the nonparametric test.Count data were analyzed by the chi-square test.The incidence of cholelithiasis in the 4 groups were calculated by the Kaplan-Meier method and comparisons of incidence were done by the Log-rank test.The univariate analysis and multivariate analysis were done using the COX regression model.Results (1) Incidence of cholelithiasis in the 4 groups:31 794 subjects were observed for (2.1 ± 0.4) years,and 236 had new-onset cholelithiasis with an incidence of 7.42‰.Incidences of cholelithiasis in the Q1,Q2,Q3 and Q4 groups were respectively 4.03‰,7.17‰,7.93‰ and 10.57‰,with a statistically significant difference among the 4 groups (x2 =72.39,P＜0.05).(2) Risk factors analysis affecting new-onset cholelithiasis:results of univariate analysis showed that sex,age,cumBMI,BMI,hypertension and CRP were independent risk factors affecting new-onset cholelithiasis of subjects [HR =1.61,1.75,1.64,1.36,1.39,1.39,95％ confidence interval (CI):1.23-2.10,1.49-2.05,1.45-1.86,1.21-1.53,1.07-1.79,1.18-1.62,P＜0.05].Results of multivariate analysis showed that female,age between 50 years and 60 years,age≥60 years,140.81 kg/m2×year ≤cumBMI ＜159.69 kg/m2×year,159.69 kg/m2×year≤cumBMI＜ 180.49 kg/m2 ×year,cumBMI ≥ 180.49 kg/m2 × year were independent risk factors affecting new-onset cholelithiasis of subjects (HR=1.59,1.78,2.33,2.04,2.42,3.66,95％CI:1.21-2.09,1.31-2.44,1.63-3.34,1.29-3.24,1.47-3.95,2.15-6.25,P＜0.05).Conclusion Female,advanced age and increasing cumBMI are independent risk factors affecting new-onset cholelithiasis,and the incidence of cholelithiasis rises as cumBMI increases.

Objective To investigate the expression levels of the CD14 mRNA and TLR4 mRNA in peripheral blood mononuclear cells from patients with Parkinson disease and to explore the clinical signicance. Methods For?ty-four patients with Parkinson disease and 37 healthy controls were recruited. We recorded age of onset, duration of ill?ness and sex of all recruited patients. PD patients were evaulated using the Hoehn-Yahr stages, Unified Parkinson Dis?ease Rating Scale (UPDRS) Ⅱand UPDRS Ⅲ,non-motor symptoms scale (NMSS) on“off”time. Reverse transcrip?tion-polymerase chain reaction was performed to determine the expression levels of CD14 mRNA and toll-like receptor 4 (TLR4) mRNA. Results The expression levels of CD14 mRNA(1.459±0.658)2-△△CT and TLR4 mRNA (1.408±0.698)2-△△CT was significantly up-regulated (P<0.05) in Parkinson disease compared with controls((1.162 ± 0.631)2-△△CT、(1.122 ± 0.557)2-△△CT). In addition, there was positive correlation between the expression levels of CD14 mRNA in Parkinson dis?ease patients with the Hoehn-Yahr stages. Meanwhile, there was no significant correlation between the expression levels of CD14 mRNA and TLR4 mRNA with other clinical scores. Conclusions There is positive correlation between the ex?pression levels of CD14 mRNA in Parkinson disease patients with the Hoehn-Yahr stages,indicating that CD14/TLR4 positive monocyte may be involved in the pathogenesis of the Parkinson disease.

OBJECTIVETo investigate the influence of different intensity and directions of ambient light and adjacent tooth in anterior tooth color measurement by using colorimeter.

METHODSFiber lite MI-150 was used as ambient illuminant and it irradiated from three or twelve o'clock direction through 45 degrees angle above. The light magnitude 0, 50, 75, 100, 125, 150 W were applied in this experiment. The values of CIE L* a* b* were measured by Minolta Chroma meter CR-321 colorimeter on the center labial surface of ten extracted human maxillary central incisors with or without adjacent teeth, then those data were analyzed statistically by using SPSS 11.5.

RESULTSNeither different intensities nor different directions of ambient light could influence the results of color measurement by using Minolta Chroma meter CR-321 colorimeter, so did the adjacent teeth whether those were exist or not.

CONCLUSIONThere is no influence of ambient light and adjacent teeth in the color measurement of anterior teeth under this experiment condition, and Minolta Chroma meter CR-321 colorimeter can be used to measure the color directly aside the chair with light.

The wind, phlegm, and blood stasis are important pathogenic factors of cough variant asthma in children, and they are also the pathological products in the occurrence and development of this disease. They have typical pathogenic characteristics. The main pathogenesis characteristics of cough caused by wind, phlegm and blood stasis are as follows that external wind attacks the lungs and induces internal wind, phlegm and dampness accumulates in the lungs, and the lungs fail to declare and descend, and blood stasis obstructs the collaterals and stagnation of Qi. The wind, phlegm, and blood stasis have their own pathogenic characteristics, and their cough-causing also have their own pathogenic characteristics and clinical characteristics. Based on the characteristics of wind, phlegm, and blood stasis, the application of medicine based on the differentiation of symptoms and signs can effectively prevent and treat this disease, and provide theoretical basis and treatment ideas for the treatment of cough variant asthma in children with Chinese medicine.

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/m 2 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], χ 2=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], χ 2=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/m 2 vs. 34.3±4.2 kg/m 2, respectively; 24.3±2.4 kg/m 2 vs. 32.7±3.7 kg/m 2, 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], χ 2=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.

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/m 2 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], χ 2=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], χ 2=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/m 2 vs. 34.3±4.2 kg/m 2, respectively; 24.3±2.4 kg/m 2 vs. 32.7±3.7 kg/m 2, 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], χ 2=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.

The rapid development of bariatric and metabolic surgery in China in the past 20 years is due to the economic development and needs of lifestyle changes, as well as the unremit-ting exploration of surgeons. Although bariatric and metabolic surgery is facing good development opportunities, its specialized development is far from enough, non-standard surgery still exists in clinical work, specialist training and standardization are in the initial stage, and follow-up is also a serious challenge for bariatric and metabolic surgery. The authors believe that on the basis of solid progress in normalization and standardization, bariatric and metabolic surgery will welcome a bright future.