Objective To assess the effects ofberberine on autophagy in C3H10T1/2 adipocytes.Methods C3H10T1/2 cells,a pluripotent stem-cell line of mesodermal origin,were induced to differentiate into mature adipocytes,and then were treated with berberine,The expression of autophagy marker protein LC3 Ⅱ/Ⅰ and autophagy substrate P62 was determined by Western blot ; after treating C3 H 10T1/2 cells with berberine and lysosomal inhibitor,chloroquine,autophagy flux was assessed by Western blot.Autophagosome was observed by transmission electron microscopy after berberine treatment.Results (1) After berberine treatment,the expression of LC3 Ⅱ / Ⅰin C3H10T1/2 adipocytes was reduced and P62 was increased in a time-and dose-dependent manner(P＜0.05 or P＜0.01) ; (2) Following treatment with chloroquine and berberine,the protein level of LC3 Ⅱ/Ⅰ was decreased (P＜0.05); (3) The number of autophagosome was decreased apparently after berberine treatment.Conclusions Berberine inhibits autophagy in C3H10T1/2 adipocytes.

Objective To compare the expressions of Beclin 1 and its 11 interacting proteins in subcutaneous fat and visceral fat,and to observe the expressions of these genes in 3T3-L1 adipocytes differentiation process.Methods In 3T3-L1 adipocytes,cells were harvested at day 0,2,4,6,8 in differentiation process,and then proteins or mRNAs were obtained and followed with Western blot and realtime PCR.Mice tissue proteins or mRNAs were detected with Western blot and realtime PCR.Results In 3T3-L1 adipocytes differentiation process,expressions of LC3,Beclin 1,and TNFAIP3 continued to rise,expression of BIRC5 was high to low,FEZ1 continued at a low level.In mice,expressions of LC3,Beclin 1,YWHAQ,FEZI,BAD,WAC,TNFAIP3,and GOPC in visceral fat were significantly higher than those in subcutaneous fat,while expression of SLAMF1was higher in subcutaneous fat.Conclusion Autophagy plays an important role in 3T3-L1 differentiation process and may be tissue-specific in visceral fat and subcutaneous fat.

Objective To investigate the clinical,endoscopic and pathologic features in the differential diagnosis between Crohn' s disease (CD) and intestinal tuberculosis (ITB).Methods The complete clinical data of 107 patients with CD and 69 patients with ITB in our hospital from January 2011 to January 2012 were retrospectively analyzed.The diagnostic value of the clinical and endoscopic scoring system was evaluated.Results CD occurred mainly in male.The salient features of CD included long duration of disease high incidence of colectomy.Comparing with patients with ITB,patients with CD have more cases of diarrhea,hematochezia,abdominal mass,intestinal obstruction,intestinal hemorrhage,perianal lesions,and extraintestinal manifestations (all P ＜ 0.05).It's more frequent to have positive results of anti-Saccharomyces cerevisiae antibody (ASCA),perinuclear antineutrophil cytoplasmic antibody (pANCA) and fecal occult blood in CD patients,as well as low albumin,high C-reactive protein (CRP),elevated platelet count and hematocrit (P ＜ 0.05 or P ＜ 0.01).The salient features of ITB included low fever,night sweats,active parenteral tuberculosis,increased erythrocyte sedimentation rate (ESR),chest X-ray abnormalities,the positive PPD (purified protein derivatives tuberculin) and T-SPOT (P ＜ 0.05 or P ＜0.01).Based on the imaging,CD often involved the small intestine,such as the intestinal stricture and abdominal abscess (P ＜ 0.05),while mesenteric lymphadenopathy was more common in ITB (P ＜ 0.05).The endoscopic examination showed that some patterns of disease involvement such as fissure-shape ulcer [41.12％ (44/107) vs 5.80％ (4/69)],cobblestone sign [15.89％ (17/107) vs 4.35％ (3/69)],lesions over four segment [24.30％ (26/107) vs 7.25％ (5/69)],rectum involvement [17.76％ (19/107) vs 5.80％ (4/69)],ileocecal valve stenosis [21.50％ (23/107) vs 8.70％ (6/69)] and mucosal bridge [5.61％ (6/107) vs 0(0/69)] were more frequent in CD patients than those in ITB patients(P ＜ 0.01 or P ＜0.05).However circular ulcers[37.68％ (26/69) vs 9.35％ (10/107)],rat-bite-like ulcers[24.64％ (17/69) vs 12.15％ (13/107)],persistent open ileocecal valves [39.13％ (27/69) vs 19.63％ (21/107)],tuberous and polypoid lesions [36.23％ (25/69) vs 20.56％ (22/107),37.68％ (26/69) vs 22.43％ (24/107)] were more common in ITB (P ＜ 0.01 or P ＜ 0.05).In terms of pathological findings,certain characteristic features such as transmural inflammation [5.61％ (6/107) vs 0 (0/69)],fissure-liked ulcers [14.02％ (15/107) vs 4.35％ (3/69)],non-caseous granulomas [5.61％ (6/107) vs 0(0/69)],lymphoid hyperplasia [16.82％ (18/107) vs 5.80％ (4/69)] and crypt abscess [9.35％ (10/107) vs 1.45％ (1/69)] were more common in CD than those in ITB(P ＜ 0.05).According to the clinical and endoscopic scoring system,the positive diagnostic rate of CD was 50.47 ％ (54/107)and of ITB was 66.67 ％ (46/69) (P ＜ 0.05).Conclusions The differential diagnosis between CD and ITB should be considered carefully based on clinical,endoscopic,pathological characteristics.The clinical and endoscopic scoring system may contribute to distinguish CD and ITB.

Calcitonin gene-related peptide( CGRP) is the strongest vasodilator discovered to date,the synthesis and release of CGRP are regulated by multiple factors.CGRP possesses multiple biological func-tions,such as vasodilator,myocardial strengthener,lung protector,brain protector,immunomodulator.It plays an important role in regulating the function in the body.

Objective:To investigate the difference of radiofrequency ablation combined with ethanol ablation and radiofrequency ablation in the treatment of benign cystic solid thyroid nodule.Methods:A total of 80 patients who visited the Thyroid Surgery Department of the First Affiliated Hospital of Zhengzhou University from January 2015 to July 2018 were selected. All selected patients are required to meet the following criteria: (1)Color doppler ultrasonography of the neck revealed a cystic solid thyroid nodule greater than 20 mm in diameter. (2) The results of fine needle aspiration cytology of thyroid nodules were benign. (3)The patients is to undergo radiofrequency ablation of thyroid nodule. According to the condition and patients′ wishes, radiofrequency ablation (Group A, n=40) and combined ethanol and radiofrequency ablation(Group B, n=40) were performed respectively to observe the changes of nodule volume and maximum diameter at 3, 6 and 12 months after surgery.The difference of intraoperative radiofrequency ablation energy, postoperative complications and patient satisfaction at 12 months after operation were also observed. The respective clinical effects of the two groups and the difference of curative effects between the two groups were analyzed. Two-factor repeated measurement analysis of variance or independent sample t test was used to compare the measurement data in line with normal distribution between groups. Friedman′s rank sum test was used for comparison of measurement data groups that did not conform to normal distribution, and Bonferroni correction was used for pairwise comparison. Chi-square test was used to compare the counting data between groups. Results:On the 12th months after operation, the volume reduction of of nodules in group B was greater than that of group A, and the difference was statistically significant[(7.0±5.1) mL vs (5.5±4.9) mL, P<0.05]. The maximum diameter reduction of nodules in group B was greater than that of group A and the difference was statistically significant [(1.5±0.6) cm vs (1.4±0.8) cm, P<0.05]. During the period of 6 to 12 months after operation, the trend of nodular shrinkage in group B was more obvious than that in group A ( P<0.05). The radiofrequency ablation energy of group was lower than that of group A, and the difference was statistically significant [(2.37±1.18) kJ vs (3.89±1.17) kJ, P<0.05]. Voice reduction occurred in 2 cases and recovered within 2 weeks.Local bleeding occurred in 1 case during the operation, which was stopped after ablation. There was no statistical significance in the satisfaction of patients in group A and group B (87.5% vs 90%, P>0.05). Conclusion:Compared with radiofrequency ablation, radiofrequency ablation combined with ethanol ablation for benign cystic solid thyroid nodules can achieve better nodule reduction effect and reduce the ablation energy.

Objective To analyze the effect of the changes of hospital diagnosis and treatment mode on the treatment time in patients with acute ischemic stroke before and after the establishment of Cerebrovascular Disease Center. Methods A total of 103 consecutive patients with acute ischemic stroke admitted to the Department of Neurology,Changhai Hospital,the Second Military Medical University between June 2008 and December 2012 were enrolled retrospectively. Thirty-one of them were excluded because of incomplete medical records. Finally,72 patients were enrolled as a control group and received series diagnosis and treatment mode. A total of 210 consecutive patients with acute ischemic stroke admitted to the Cerebrovascular Disease Center,Changhai Hospital,the Second Military Medical University from September 2013 to February 2015 were enrolled retrospectively. Thirteen patients were excluded (4 patients with recurrent transient ischemic attack were treated with recombinant tissue-type plasminogen activator,9 without complete data were treated with intravenous thrombolysis),197 were enrolled as an observation group finally,and they were received series diagnosis and treatment mode. The patients of both groups were visited within 4. 5 h after onset and received rt-PA treatment. The time-consuming changes of each time period from onset-to-door,door-to-imaging,imaging-to-needle,door-to-needle,and onset-to-needle time between the control group and the observation group were compared and analyzed. Results Compared with the control group,the door-to-imaging,imaging-to-needle,door-to-needle and onset-to-needle time were significantly shorter in the observation group. There were significant difference between the 2 groups (24 ± 12 min vs. 60 ± 20 min,27 ± 12 min vs. 62 ± 31 min,51 ± 17 min vs. 122 ± 52 min,and 153 ± 69 min vs. 230 ± 81 min,all P < 0. 01). There was no significant difference for onset-to-door time between the observation group and the control group (P > 0. 05). Conclusion The establishment of cerebral vascular disease center and the improvement of the processes have shortened the treatment time in patients with acute ischemic stroke within time window. The time from onset-to-door is still longer,and the propaganda and education of stroke should be strengthened.

Objective To investigate the effect of adenovirus-bone morphogenic protein 9 ( Ad-BMP9 ) on osteogenic differentiation of immortalized calvarial mesenchymal progenitor cells ( iCALs ) .Methods iCALs were infected with adenoviral vectors encoding BMP-9 or green fluorescent protein ( GFP) and the early osteogenic differentiation was assessed by detecting alkaline phosphatase (ALP) activity after being cultured for 3, 5 and 7 days.14 days after infection, alizarin red S staining was performed to study the formation of osteogenic calcium nodules .The expression of osteogenic marker genes Runx2 and OCN was assessed by quantitative real-time ( RT )-PCR and Western blotting .Results Significant increases in ALP activity and in the expressions of Runx 2 and OCN were detected in BMP-9 treated iCALs compared with GFP-treated cells(P<0.05).Meanwhile, alizarin red S staining showed that more mineralized nodules were found in the BMP-9 induced group .Conclusion BMP-9 can promote the osteogenic differentiation of iCALs .

Objective: To observe the relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia and evaluate the safety preliminarily.Methods: The relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia were studied by the cough model caused by the irritation of ammonia water and the phenol red output of trachea in mice.The acute toxicity test and maximum tolerance test were carried out to evaluate the safety.Results: The total alkaloid in Atalantia buxifolia at low dose could obviously prolong cough incubation period and decrease cough times in mice, and that at high dose could significantly increase the secretion of phenol red in respiratory tract, and compared with those in the blank group, the differences were statistically significant (P<0.05).In the acute toxicity test, no death showed after the administration with maximum tolerance dosage, and the rate of weight growth had no difference between the blank group and the model group (P>0.05).Conclusion: The relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia are notable in the cough model caused by the irritation of ammonia water and the phenol red output of trachea in mice.The maximum tolerable dose test shows the total alkaloid in Atalantia buxifolia is safe.

Objective:To explore the possibility of using peritoneal alternatively activated M2 macrophages to prevent rejection after islet allotransplantation in a murine model.Methods:Peritoneal monocytes from C57BL/6 mice were induced and modulated to M2 and M0 macrophages in vitro,then the phenotype of macrophage was assessed by flow cytometry.C57BL/6 mice were induced diabetic by streptozotocin (STZ) injection and transplanted with islets isolated from BALB/c mice under the left kidney capsule.The recipients were randomly divided to 3 groups (n=8).A total of 2.5× 106 M2 macrophages were injected intravenously at 0,3,7 d after transplantation in islet+M2 group;2.5×106 M0 macrophages were injected intravenously at 0,3,7 d after transplantation in islet+M0 group;the mice in islet+PBS group were injected with PBS.Blood glucose was monitored after transplantation.On day 10 after transplantation,2 recipients in each group were randomly selected and sacrificed,and the left kidneys were resected for pathological examination.Results:Achievement of euglycemia was significantly prolonged after islet transplantation in the islet+M2 group than that in the other two groups (P＜0.01).The median survival time of islet allografts in the islet+PBS group,the islet+M0 group,and the islet+M2 group were 6.5 (4-10),7.5 (4-10),and 24(＞ 15) d,respectively.Pathological examination also showed that the grafts in islet+M2 group remained an intact structure with positive insulin stain and no apparent lymphocytes infiltration,while the graft was rejected in other 2 groups with negative insulin stain and massive lymphocytes infiltration.Conclusion:Peritoneal alternatively activated M2 macrophages can prevent rejection after islet allotransplantation,prolong the survival time of islet allografts and enhance the tolerance of the recipient to blood glucose in mice.

Aim To investigate the neuroprotective effect of verbascoside,one of the phenylethanoids isolated from the stems of Cistanche salsa,on MPP+-induced injury in SHSY5Y cells.Methods SHSY5Y cells were exposed to various doses of verbascoside for 12 h,and then treated with 200 ?mol?L-1 MPP+ for 24 h.The cell viability was observed with MTT assay;reactive oxygen species,the mitochondrial membrane potential and the percentage of apoptosis were measured by flowcytometer;the activation of caspase-3 was measured with the caspase-3 activity assay kit;the expression of Bcl-2 was measured with Western blot.Results Following treatment with MPP+ for 24 h,MPP+ induced a significant decrease of cell viability;apoptosis percentage were 38.9%;accumulation of intracellular ROS,increase of caspase-3 activity and the decreases in mitochondrial membrane potential were detected.However,pretreatment with verbascoside (0.1,1 or 10 mg?L-1) for 12 h exhibited cytoprotective effects in a dose-dependent manner.Verbascoside obviously enhanced cell viability,and significantly reduced the number of cells labeled with Annexin-V.The percentage of apoptosis neurons was significantly decreased to 29.5%,15.3% and 8.6% respectively.Flowcytometer showed the verbascoside attenuated the accumulation of ROS and the MPP+-induced collapse of mitochondrial membrane potential in SHSY5Y cells.And significant decreases were detected in caspase-3 activity compared with the MPP+-treated cells at the same time point.Moreover,pretreatment with verbascoside promoted the expression of Bcl-2.Conclusions verbascoside had the neuroprotective capacity to antagonize MPP+-induced apoptosis in SHSY5Y cells,and might be useful in treating Parkinson disease.