Objective To investigate the possibility of transfection of adipose-derived mesenchymal stem cells (ADMSCs) into the insulin-secreting cells in vitro,and assay the secretion of insulin of ADMSCs in high and low glucose environment.Methods The ADMSCs that untransfected were in the control group,the ADMSCs that contained PcDNA3.1 were in the vacant vector group,and the ADMSCs that contained PcDNA3.1-hINS were in the recombinant vector group.After transfection,the recombinant vector group were sub-divided into the 1,6,12,18 days groups.According to the concentrations of glucose,the recombinant vector 18 days group were divided into the high glucose group and low glucose group.Human insulin gene was amplified by RT-PCR,and the eukaryotic expression recombinant vector PcDNA3.1-hINS that contained the human insulin gene was constructed.The ADMSCs were obtained by digestion and centrifugation,and then underwent flow cytometry for identification.The transcription of insulin DNA was assayed by RT-PCR,and the levels of insulin were assayed by ELISA.Glucose test was done in the recombinant vector 18 days group.The measurement data was shown in the format of (x) ± s,the measurement data in multiple groups were compared by randomized analysis of variance,and the comparison among groups was performed by the t test.ResuIts The expressions of CD44,CD90,CD106 were positive,and the expressions of CD34,CD45 and CD11b were negative.No insulin DNA transcription was detected in the control group and vacant vector group.The levels of insulin secreted were (4.7 ± 0.8) mIU/L,(8.8 ± 0.5) mIU/L,(8.9 ± 0.8)mIU/L,(8.6 ± 0.6)mIU/L in the recombinant vector 1,6,12,18 days group,which were significantly higher than (1.3 ± 0.6) mIU/L in the control group and (1.7 ± 0.8) mIU/L in the vacant vector group (t =10.09,32.64,22.20,55.53 ; 9.23,27.56,19.43,51.25,P ＜ 0.05).There were significant differences in the levels of insulin secreted between the recombinant vector 1 day group and the recombinant vector 6,12,18 days groups (t =12.77,12.26,13.93,P ＜0.05).There were no significant difference in the levels of insulin secreted between the recombinant vector 6,12,18 days groups (F =45.67,P ＞ 0.05).There was a significant difference in the level of insulin secreted between the high glucose group and the low glucose group (t =2.03,P ＜ 0.05).The result of the glucose stimulation test was negative.Conclusion The ADMSCs are transfected into insulinsecreting cells in vitro successfully,and the secretion of insulin is stable.Although the secretion of insulin can't change in line with the concentration of glucose,it is a new seed cell for the treatment of diabetes with stem cells.

Objective:To analyze the benefit incidence and evaluate the implementation effects on catastrophic disease insurance under the New Rural Cooperative Medical System ( NRCMS) in Jilin Province and put forward sug-gestions to perfect the catastrophic disease insurance under NRCMS. Method: In-depth interviews with related staff were conducted to gain an understanding of the operating status of catastrophic disease insurance under NRCMS;cat-astrophic disease patient data of Jilin Province from 2013 were collected, with a comparative analysis of implementa-tion effects among nine districts;the benefit incidence was analyzed by means of the Sperman rank correlation analy-sis; Results: The policy of catastrophic disease insurance under the NRCMS exhibited sound implementation effects and played an important role in reducing the economic burden of catastrophic disease patients. The average individual expenditure decreased by 4336. 52 yuan, while the reimbursement rate increased by 11. 15% after NRCMS cata-strophic disease insurance reimbursements. Additionally, statistical analysis results showed that there was a weak cor-relation between benefit incidence and economic level, the benefit equality of catastrophic disease insurance under NRCMS was relatively good, but the benefit level of rich patients was slightly higher. Conclusion:The reimbursement scheme of catastrophic disease insurance under NRCMS should be refined and improved, and the benefit fairness of different districts must be ensured, thus achieving the goal of relieving the“poverty caused by illness” phenomenon.

A total of 52 patients with chronic lateral epicondylitis were randomly assigned into platelet-rich plasma (PRP) (n =28) or corticosteroid (n =24) group.The visual analog pain scale (VAS) and Mayo elbow score had no significant difference between two groups within 1 week post-treatment.However,VAS and Mayo elbow score improved more significantly in PRP group versus corticosteroid group at Month 1 and 6 post-treatment (P ＜ 0.05).The PRP trigger point injection treatment of elbow lateral epicondylitis achieved clinical outcomes and it was superior to glucocorticoid treatment.

AIM: To explore the relationship between caspase activation and the evasion of Legionella from macrophage elimination through a Legionella-infected macrophage model. METHODS: After infected by Legionella, the activity of caspase 3 in macrophages was analyzed by confocal microscopy as well as fluorescence reader. Growth and replication of Legionella in macrophage was assayed. Replication of Legionella was analyzed again to see the effect of caspase 3 inhibition on the growth of Legionella after use of caspase 3 inhibitor. RESULTS: Both confocal microscopy and caspase 3 fluorescent substrate analysis showed that Legionella virulent strain had powerful capability of activating caspase 3 while the mutant non-virulent strain did not have this capability. The virulent strain highly replicated in macrophages and the replication was significantly inhibited by caspase 3 inhibitor. CONCLUSION: Our results indicate that the intracellular caspase 3 is activated shortly after infection by Legionella virulent strain. The evasion of Legionella from the elimination of macrophages may be mediated by caspase 3 activation to a great degree.

Objective To dynamically monitor the base excess(BE) in traumatic brain injury(TBI) patients within 3 d after admission,and to assess the impact of the early BE on prognosis.Methods Blood BE was monitored for 3 d in 56 TBI patients.Patients were classified into mild group(15 patients),moderate group (22 patients) and severe group (19 patients) according to the scores of Glasgow coma scale(GCS).Patients were classified into survival group(42 patients) and dead group(14 patients) according to prognosis.Patients were classified into high BE group (35 patients,BE≥-8 mmol/L) and low BE group (21 patients,BE ＜-8 mmol/L).The relations among BE,degree of injury and prognosis were analyzed.Results The level of BE in mild group,moderate group and severe group was increased after treatment for 1,2,3 d than that before treatment.The level of BE was consistent with the degree of injury [mild group:(-3.02 ± 0.21)mmol/L; moderate group:(-8.49 ± 1.44) mmol/L;severe group:(-9.64 ± 1.19) mmol/L].The level of BE in mild group and severe group had significant difference than that in moderate group (P＜ 0.01).The level of BE in dead group before treatment and after treatment for 1,2,3 d was significantly lower than that in survival group [(-11.97 ±2.13) mmol/L vs.(-6.29 ± 1.16) mmol/L,(-9.84 ± 1.33) mmol/L vs.(-4.89 ± 1.78)mmol/L,(-8.78 ± 2.01) mmol/L vs.(-3.61 ± 1.43) mmol/L,(-7.84 ± 1.42) mmol/L vs.(-3.10 ±0.98)mmol/L] (P ＜0.01).The scores of APACHE Ⅱ before treatment and fatality rate in low BE group were significantly higher than those in high BE group [(24.84 ± 3.68) scores vs.(16.27 ± 2.21) scores,52.4％ (11/21) vs.8.6％(3/35)] (P ＜ 0.01).The scores of GCS before treatment in low BE group was significantly higher than that in high BE group [(7.56 ± 3.09) scores vs.(10.51 ± 2.43) scores](P ＜ 0.01).Conclusion The level of early BE is a good factor on evaluating the condition and prognosis in TBI patients.

AIM: To explore the effect of TNF-related apoptosis inducing ligand (TRAIL), a new apoptotic inducing molecule on the biological activity of hepatocarcinoma cell line. METHODS: The expression of membrane binding TRAIL on HepG2 cells was detected by immuno-cytochemistry. Quantity of secretory TRAIL was assayed by ELISA method. The cytotoxicity and apoptosis induced by TRAIL was detected by MTT and TUNEL method, respectively. The telomerase activity of HepG2 cells was detected by TRAP-PCR assay kit. The expression of hTERT, the catalytic subunit of telomerase, was detected by FCM. RESULTS: TRAIL was constitutively expressed on the membrane of HepG2 cell line. Soluble TRAIL was also expressed to a certain degree. Cytotoxicity assay showed that TRAIL significantly inhibited the growth of hepatocarcinoma cells. TUNEL assay indicated that TRAIL induced apoptosis in hepatocarcinoma cells. Detection of telomerase activity showed that TRAIL inhibited telomerase activity and the expression of telomerase catalytic subunit. CONCLUSION: TRAIL is an effective molecule to inhibit the growth of hepatocarcinoma through multiple pathways, such as inducing apoptosis and inhibiting the activity of telomerase.

[Summary] Sixty-one patients suffering from pituitary apoplexy( PA) were mainly diagnosed according to pathologic findings, and were collected from case record, pathology, and MRI databases. They were classified into 4 types according to the clinical condition: the insidious type was characterized with only positive pathological findings;the asymptomatic type had both positive pathologic and MRI findings; the subacute type had PA associated symptoms longer than 2 weeks; and the acute type had PA associated symptoms for 2 weeks or less. The latter 2 types had positive pathological and MRI findings additionally. The basic lesions, acute or chronic symptoms, endocrinopathies and MRI findings were compared among 4 types. Results showed as followed. In all patients, there were headache(60. 7% ), blurred vision(55. 7% ), vomiting(21. 3% ), and dizziness(14. 8% ). Apoplexy associated symptoms comprised severe headache (24. 6% ), rapid vision loss (29. 5% ), and blepharopotosis or diplopia (9. 83% ). Insidious, asymptomatic, subacute, and acute types were composed of 15 (24. 6% ), 9 (14. 8% ), 19 (31. 1% ), and 18 (29. 5% ) cases, respectively. Aging and intracranial space-occupying symptoms as first complaint showed increasing trend from mild to severe types(both P<0. 05), while in chronic course it showed decreasing trend(P<0. 05). Acute massive symptoms(P<0. 01), and non-functional tumor(P<0. 01) in the 2 clinical types were much more frequent than in the two mild types. Half or more pituitary-target glands showed impaired functions in each type, and the impairment showed increasing trend through mild to severe types(P<0. 01). The present study provided a brief typing system in order to expand PA concept to a wider span covering various conditions. Some differences in tumor composition and endocrinopathies existed among the four types.

Objective Objective To explore the best dog urethra rectum fistula operation method.Methods In June 2016 to October 2016,15 healthy male dogs were selected as research objects,aged 2-3 years (mean 2.5 years),with a body mass of 12-15 kg (mean 13.5 kg).They were randomly divided into three groups,each group of five.5 male dogs were cut through the rectum.5 male dogs were cut through the perineum.5 male dogs were cut through the rectal sphincter of the anus.Postoperative retrograde urethrectomy rectum angiography,the urethral pressure injection of methylene blue white gauze aizen rectum fistula and histologic evaluation of each group were compared 1,2,3 moths after the operation.Results Anal rectum drag cold knife cut group,1,2,3 moths after the operation,postoperative indicated the exist of orificium fistulae.The urethral pressure injection of methylene blue could observe rectum white gauze aizen,and pathological histology suggested the mucosal lamina propria disorder.There were a large number of inflammatory cells,5 dogs models were successfully built.Abdominal perineal pathway group:1 dog was killed by itself.1 dog one month after operation in anus at 2 cm below diameter about 3 cm appeared in a mixture of urine fistula and urethral orifice outflow,no urine outflow.More than 3 dogs after a week in the edge of serious infection,a canine model of severe dysuria,1,2,3 moths after the operation,postoperative indicated the exist of orificium fistulae.The urethral pressure injection of methylene blue could observe rectum white gauze aizen.3 dogs models were built.Through anal sphincter incision group:five dogs of the urethral fistula were successful,and one dog was incontinent with fecal incontinence.Conclusions The rectal rectum could successfully produce the animal model of the urethral rectal fistula and provide the experimental subjects for the treatment of iatrogenic urethral fistula.

Objective To investigate the relation between fragmentation of the Golgi apparatus and expression changes of Rab30 in HT22 neurons.Methods The routinely cultured HT22 neurons were divided into control group,thapsigargin 3 h group and thapsigargin 5 h group;neurons in the thapsigargin 3 h group and thapsigargin 5 h group were given 0.1 μg/mL thapsigargin treatment for 3 or 5 h,respectively.Morphology of Golgi apparatus and location of Rab30 were observed with cytochemistry immunofluorescence.The Rab30 mRNA expression was detected by real-time fluorescent quantitative PCR.The protein expressions ofRab30,cis-Golgi matrix protein 130 (GM130),and golgi anti-apoptotic protein (GAAP) were detected by Western blotting.Results Thapsigargin induced markedly Golgi fragmentation in HT22 neurons.The longer the neurons were exposed to thapsigargin,the more the Golgi fragmentation.Hippocampal neurons also expressed Rab30,which was mostly localized on Golgi apparatus.The Rab30 mRNA and protein expressions in the control group,thapsigargin 3 h group and thapsigargin 5 h group were decreased in sequence (relative transcript level:1.00±0.00,0.73±0.05 and 0.53±0.01),with significant differences (P＜0.05).The GM130 and GAAP protein expressions in control group,thapsigargin 3 h group and thapsigargin 5 h group were decreased in sequence,with significant difference (P＜0.05).Conclusion Thapsigargin can induce Golgi fragmentation,which may be related to the decreased expression of Rab30 in HT22 neurons.

ObjectiveTo explore the possible mechanism of Yiqi Yangyin Huoxue prescription in the prevention and treatment of kidney injury of diabetic kidney disease（DKD）rats based on NOD-like receptor protein 3（NLRP3）/cysteine protease-1（Caspase-1）/gasdermin D （GSDMD）pyroptosis pathway. MethodFifty male SD rats were randomly divided into normal group （n=8） and modeling group （n=42）. The modeling group was given a one-time intraperitoneal injection of streptozotocin （STZ） after high-sugar and high-fat diet for 6 weeks to induce the establishment of a DKD rat model. After successful modeling， the rats were randomly divided into model group， valsartan group （8.33 mg·kg^-1）， and Yiqi Yangyin Huoxue prescription low-dose and high-dose group （11，22 g·kg^-1）. After continuous gavage for 6 weeks， the fasting blood glucose （FBG）， total cholesterol （CHO）， triglyceride （TG）， blood urea nitrogen （BUN）， serum creatinine （SCr） and 24-hour urine protein quantification （24 h-UTP） were detected in each group of rats. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of kidney tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum interleukin-1β （IL-1β） and interleukin-18 （IL-18） levels. The protein and mRNA expression levels of NLRP3/Caspase-1/GSDMD in kidney tissue of rats in each group were determined by Western blot and real-time quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the conditions in normal group， the levels of FBG， CHO， TG， BUN， SCr， 24 h-UTP and serum IL-1β and IL-18 as well as the protein and mRNA expression levels of NLRP3/Caspase-1/GSDMD in kidney tissue in model group were increased （P<0.01）， and the kidney tissue lesions were severe. Compared with the conditions in model group， the levels of FBG， CHO， TG， BUN， SCr， 24 h-UTP and serum IL-1β and IL-18 as well as the protein and mRNA expression levels of NLRP3/Caspase-1/GSDMD in kidney tissue in each intervention group were decreased （P<0.05， P<0.01）， and the degree of kidney tissue lesions was improved， with Yiqi Yangyin Huoxue prescription high-dose group showing the optimal effect. ConclusionYiqi Yangyin Huoxue prescription could inhibit pyroptosis by regulating the NLRP3/Caspase-1/GSDMD pathway， and thus relieve the inflammatory response of DKD rats and alleviate the pathological damage of the kidneys.