The level of serum fibroblast growth factor-21 (FGF-21) in patients with type 2 diabetes mellitus complicated with non-alcoholic fatty liver disease (NAFLD) was determined by ELISA.The results showed that serum FGF-21 level in these patients was higher than that in type 2 diabetic patients [(266.55 ± 21.24 vs 220.32 ± 22.68) ng/L,P＜ 0.01].Serum FGF-21 levels in both groups were significantly higher than that in normal control group [(173.52 ± 16.18) ng/L,P＜0.01].Serum FGF-21 level was positively correlated with waist circumference,blood glucose,and triglyceride.FGF-21 may contribute to the development of NAFLD in the patients with type 2 diabetes mellitus.

The changes of serum apelin in patients with metabolic syndrome were investigated. The level of fasting serum apelin was determined by ELISA assay.The level of serum apelin was raised obviously in patients with metabolic syndrome(MS).Moreover,the level of serum apelin in patients with type 2 diabetes mellitus was higher than those in cases of MS with impaired glucose regulation and the group of subjects with overweight/obesity [ (475.8±37.3 vs 451.5±54.3 and 430.3±52.1 ) ng/L,P＜0.01 ].In a multiple step-wise regression analysis,In homeostasis model assessment of insulin resistance ( HOMA-IR),body mass index,and total cholesterol were independent factors of apelin.The increased serum apelin was closely related to metabolic abnormalities in metabolic syndrome.The increased insulin resistance might cause the raised level of serum apelin in patients with MS.

Objective To study the effects of Apelin on glucose toxicity and islet cells PDX-1 protein expression.Methods The islet β cell line NIT-1 cells were incubated in the medium containing different glucose concentrations(normal glucose concentration group 5.6 mmol/L,high glucose concentration group 16.7 mmol/L,extremely high glucose concentration group 33.3 mmol/L) and +/-Apelin-36 respectively for 3 d.Then the basic insulin secretion amount of islet cells and their secretion amount after glucose stimulation were detected.The intracellular insulin content and the PDX-1 protein and mRNA expression were detected.Results Compared with the normal glucose group,the basic insulin secretion,secretion after stimulation and intracellular insulin in the high glucose group and extremely high glucose group were significantly decreased and PDX-1 protein expression was declined(P＜ 0.05);compared with non-adding Apelin group,the basic insulin secretion,secretion after stimulation and intracellular insulin in the adding Apelin high glucose group and extremely high glucose group were significantly decreased and PDX-1 protein expression was decreased(P＜0.05);the insulin level in islet cells of 6 groups was positively correlated with PDX-1 protein expression and had no correlation with PDX-1 mRNA expression.Conclusion Apelin may participate in the glucose toxic effect by decreasing PDX-1 protein expression,causes the decrease of insulin secretion,thus plays a role in the pathogenesis of diabetes.

Objective To establish a model for preparation of tissue-engineered skin grafts with hTERT cells carrying human papillomavirus type 6 (HPV 6) genome in vitro, so as to lay a foundation for studying HPV life cycle. Methods The full-length linear HPV6 genome and plasmid pEGFP-▲EGFP were electrophoretically cotransferred into hTERT cells. After selection using G418 resistance, Southern blotting was performed to determine the viral load of HPV6 in transfected cells. 3T3 J2 trophoblastic cells, type I rat-tail collagen and hTERT cells containing the full-length HPV6 genes (HPV6.hTERT cells)were mixed and cocultured on metal meshes to form skin graft-like structures. Hematoxylin and eosin (HE)staining was performed to observe the structure of formed skin grafts, an immunohistochemical assay to measure the expression of HPV6 L1 protein, and electron microscopy to observe virus particles in the skin grafts. Results The linear HPV6 gene was successfully transferred into hTERT cells, and Southern blotting showed the presence of HPV6 DNA in the transferred hTERT cells. The HPV6.hTERT cells, which were cocultured with 3T3 J2 trophoblastic cells and type I rat-tail collagen, proliferated and differentiated over time, and gradually formed skin grafts giving the appearance of verrucous hyperplasia. HE staining showed that the cocultured HPV6.hTERT cells could form typical stratified structure of skin after 7 days of cultivation, and histopathologic features of HPV infection, including obvious papillomatous hyperplasia, presence of vesicular cells, hyperkeratosis and parakeratosis, could be observed after 21 days. The immunohistochemical assay showed the expression of HPV6 L1 protein in the upper portion of skin grafts, and electron microscopy revealed the presence of HPV6 virus particles in skin grafts. Conclusions The established model for preparation of tissue-engineered skin grafts using HPV 6 genome-carrying cells provides a basis for biological studies of HPV, but its application is limited to some degree.

Objective:To explore the effect of laparoscope combined with choledochoscope in the treatment of cholecystolithiasis complicated with choledocholithiasis.Methods:Totally 40 patients with cholecystolithiasis complicated with choledocholithiasis in the Second People's Hospital of Anqing from January 2016 to December 2019 were divided into double mirror combined group (20 cases) and open surgery group (20 cases) according to the operation mode.The patients in the double mirror combined group were treated by laparoscope combined with choledochoscope, and patients in the control group were treated by cholecystectomy and common bile duct exploration.The operation time, amount of bleeding, time to first flatus, postoperative complications and blood indicators were compared between the two groups.Results:The amount of bleeding[(32.50±14.82)mL], the length of hospital stay[(17.30±3.34)d], and the incidence of complication[ 5%(1/20)] in the double mirror combined group were lower than those in the open surgery group[(68.50±30.82)mL, (21.15±5.18)d, 40%(8/20)], the operation time[(162.50±56.39)min] in the double mirror combined group was longer than that in open surgery group[(102.25±21.18)min], there were statistically significant differences ( t=4.707, 2.792, 4.473, χ 2=2.692, all P<0.05). The levels of alanine aminotransferase (ALT), total bilirubin (TBIL) and albumin (ALB) in the two groups after operation were lower than those pre-operation, which in the double mirror combined group were better than those in the open surgery group[double mirror combined group (preoperation and postoperation): ALT(215.35±272.00)U/L, (44.60±44.63)U/L, TBIL(38.80±43.23)μmol/L, (16.68±11.93)μmol/L, ALB(42.65±3.25)g/L, (39.58±3.78)g/L; open surgery group(preoperation and postoperation): ALT(201.78±61.14)U/L, (61.14±48.35)U/L, TBIL(80.89±91.16)μmol/L, (40.24±53.61)μmol/L, ALB(39.37±6.81)g/L, (34.11±4.78)g/L], there were statistically significant differences ( t1=2.86, 2.72, 4.02; t2=3.54, 3.89, 4.34, t3=3.56, 4, 12, 4.01, all P<0.05). There was no statistically significant difference in white blood cell count ( P>0.05). Conclusion:The clinical efficacy of laparoscope combined with choledochoscope in the treatment of cholecystolithiasis complicated with choledocholithiasis is better than open surgery.

This study was aimed to develop in-line monitoring of extraction process of Lonicerajaponica in the extraction process trajectory of Re-Du-Ning (RDN) injection.Several batches of near-infrared (NIR) spectrum dates were collected and multivariate statistical process control (MSPC) method was in conjunction with.The results showed that using score,Hotelling T2 and DModX control chart,various normal and abnormal behaviors of the test batches were detected in time by comparison with the extraction process trajectory.It was concluded that the process trajectory for in-line quality control based on NIR spectrum dates and MSPC could indicate the changes of process.It was a feasible technology tool of the total process quality control during traditional Chinese medicine (TCM) manufacturing process.

Objective:To prepare the resiquimod-loaded lipid microbubbles R848-MBs, evaluate their enhanced ultrasound imaging and high intensity focused ultrasound (HIFU) ablation effects, and explore their ability to improve tumor immune microenvironment synergize with HIFU.Methods:R848-MBs were prepared by the thin film hydration-mechanical shock method; The basic characteristics and safety of R848-MBs were detected, the HIFU controlled-release characteristics were verified in vitro and the drug metabolism and biological distribution were investigated in vivo. The ability of enhancing ultrasound imaging was observed in vitro and in vivo. To investigate the enhanced HIFU ablation effect of R848-MBs, six EMT6 tumor-bearing mice were randomly divided into HIFU group and R848-MBs+ HIFU group, three mice in each group, the changes in contrast average sound intensity before and after ablation in mouse tumor areas and the change of ultrasound image gray value in tumor area were evaluated, the tumor were resected to observe the coagulative necresis by TTC staining and HE staining. Forty-five tumor-bearing mice were randomly divided into control group, Free R848 group, HIFU group, Blank-MBs+ HIFU group and R848-MBs+ HIFU group, nine mice in each group. On the third day after treatment, 3 mice in each group were randomly selected and killed, to evaluate the ability of R848-MBs to improve tumor immune microenvironment synergize with HIFU. The expression level of CRT on the surface of tumor cells were detected by immunofluorescence staining, the proportion of mature DC in lymph nodes, spleen, and CD8 + T cells in spleen were detected by flow cytometry. The treatment effectiveness of each group( n=6) were evaluated by measuring tumor volume, observing and drawing survival curves. Results:The R848-MBs lipid microbubbles with good safety were successfully prepared, with a concentration of 2.58×10 9/ml, as spherical bubbles under optical microscope and laser confocal microscopy, in a particle size of (1.72±0.11)μm, at a surface potential of (-10.16±0.73)mV. The cumulative drug release was up to 83.44% after HIFU (90 W, 3 s) in vitro. The concentration of R848 in plasma decreased rapidly, and the drug concentration in tumor tissue of the R848-MBs+ HIFU group was higher than that of the R848 group 24 hours after treatment ( P<0.01). The ultrasound imaging of R848-MBs was significantly enhanced in contrast mode in vitro and in vivo; R848-MBs can significantly enhance the HIFU ablation effect, the contrast average sound intensity change in the tumor area before and after ablation in the R848MBs+ HIFU group was greater than that in the R848 group ( P<0.05), and the immediate ultrasound grayscale value change in the HIFU+ R848-MBs group was 46.34±3.21, which was significantly greater than that in the HIFU group (10.67±1.53), with statistical significance ( P<0.000 1). Coagulation necrosis was observed in tumor HE staining and TTC staining. The results of treatment efficacy in vivo showed that R848-MBs+ HIFU group had the strongest therapeutic effect, and R848-MBs combined with HIFU treatment could significantly prolong the survival period of mice compared with intravenous injection of free R848 ( P<0.01). Immunofluorescence staining and flow cytometry results showed an increase in the expression level of CRT on the surface of tumor tissue in the R848-MBs combined with HIFU group, and the percentage of mature DC in tumor draining lymph nodes (58.53±1.04)% were significantly higher than those in the HIFU group (37.56±2.13)% ( P<0.001), and the percentage of mature DC in the spleen (70.65±1.91)% were significantly higher than those in the HIFU group (36.46±3.89)% ( P<0.001), the percentage of CD8 + T cells in the spleen (27.46±3.04)% was significantly higher than that in the HIFU group (18.69±0.29)% ( P<0.01). Conclusions:The HIFU controlled-release lipid microbubbles R848-MBs can not only enhance the efficiency of HIFU ablation, but also improve the tumor immune microenvironment.

OBJECTIVE To explore the improvement effect of rape pollen on androgenic alopecia mice and its mechanism. METHODS The blank group, model group, positive drug group and administration group were set up, the androgenic alopecia mice model was induced by applying 0.2% testosterone after hair removal. The hair growth rate of mice were observed by using 5% minoxidil as positive drug and 0.4 g·mL-1 rape pollen oil solution as administration group. The hair quality and follicle condition of mice were observed by scanning electron microscope(SEM) and HE staining of skin tissue, respectively. The level of VEGF and bFGF in skin were detected by immunofluorescence staining and Western blotting, while the level of serum sex hormones and reactive oxygen species were detected by ELISA. RESULTS Rape pollen could significantly promote the hair growth in mice and improve the state of mice hair scales compared with model group. Mechanism exploration experiments showed that rape pollen could not promote hair regeneration of mice by regulating hormone levels or anti-oxidative stress. However, rape pollen could increase the expression of bFGF and VEGF related to skin angiogenesis at the modeling site. CONCLUSION Rape pollen can promote hair regeneration in androgenic alopecia mice. Its mechanism may be that it promotes perifollicular vascular regeneration by increasing bFGF and VEGF level.