Objective To construct the recombinant plasmid carrying human sTNFR1 cDNA, and express sTNFR1 in E. Coli JM109. Methods Total RNA was extracted from Hela cells, and used as a template to amplify human sTNFR1 cDNA by RT-PCR. The PCR products were cloned into T vector, and then sTNFR1 cDNA fragment was subcloned into a prokaryotic expression plasmid pMAL-c2x. The recombinant plasmid was transferred into E. Coli JM109, and induced by IPTG to express fusion protein sTNFR1-MBP. sTNFR1-MBP was purified by amylose resin affinity chromatography(ARAC), and analyzed by SDS-PAGE. Results A 558 bp human sTNFR1 cDNA was amplified by RT-PCR, and successfully inserted into plasmid pMAL-c2x. sTNFR1-MBP was produced in E.Coli after IPTG induction, and a 66 KD sTNFR1-MBP was purified by ARAC. [WTHZ]Conclusion Recombinant plasmid carrying human sTNFR1 cDNA was successfully constructed and epxressed in E. Coli JM109.

Objective To construct an eukaryotic expression vector of human sTNFR1 and to investigate its inhibitory effects to the bioactivity of TNF-?.Methods The total RNA was extracted from HeLa cells and used as a template to amplify human sTNFR1 gene by reverse transcription polymerase chain reaction(RT-PCR).The PCR products were cloned into T vector and sub-cloned into vector pcDNA3.1(-),an eukaryotic expression vector.The recombinant plasmid pcDNA3.1(-)-sTNFR1 was transfected into QSG7701 cells by using lipofectamine,RT-PCR was performed to detect the expression of sTNFR1,MTT was used to observe sTNFR1 gene 's inhibitory effect on TNF-?.Results QSG7701 has a higer expression level of sTNFR1 mRNA than pcDNA3.1(-) trancfected control.The cytotoxic effect of TNF-? was inhibited to the extent of 64.8% when its concentration was 100 ?g/L.Conclusion We constructed the eukaryotic expression vector containing human sTNFR1 gene and the cytotoxicity of TNF-? is inhibited in pcDNA3.1(-)-sTNFR1/QSG7701 cells.

Objective To observe and evaluate the effects of the deep hypothermic circulatory arrest(DHCA) and regional cerebral perfusion(RCP) in pediatric aortic arch surgery.Methods According to different methods of CPB,70 infants less than 3-month-old with CoA or IAA were undergone corrective surgery with DHCA or RCP.The bypass time,aortic clamp time,DHCA or RCP time,ventilation time,ICU stay time and post-operative complications were recorded and compared between two groups.Results The incidence of neurological complications was significantly higher in DHCA group.The CPB time was significantly longer in the RCP group,and the RCP time was significantly longer than DHCA time.Blocking time,ventilator intubation time,ICU residence time,postoperative renal dysfunction,low cardiac output,puhnonary inflammation and hospital mortality was no significant difference between the two groups.Conclusion RCP is an effective cerebral protection technique.Compared with DHCA,RCP works better in sustained brain cerebral perfusion and is suitable for complex aortic arch operation in children.It has a better effort in protection of the neurological system than DHCA.

Purpose To investigate the clinicopathologic features of cervical adenoid basal cell carcinoma. Methods Clinical and pathological data were analyzed in 12 cases of cervical adenoid basal cell carcinoma and the relevant literatures were reviewed. Results The age of 12 cases of with cervical adenoid basal cell cancer patients ranged from 36~70 years ( mean:53. 3 years) , with a medi-an age of 51. 5 years. Amongst 12 patients who contained no gross definite lesion, 5 patients (41. 7%) had the symptoms of vaginal bleeding. 8 cases of patients underwent hysterectomy, while the other four received cervical conization. Cervical intraepithelial neopla-sia ( CIN) lesions were observed in all 12 cases. The tumor cells were small and uniform, with dark oval nuclei without conspicuous nucleoli and scanty cytoplasm. Tumor infiltrated into the stroma in nests and cords. Glandular differentiation within tumor nests were seen in 12 cases, including two cases of squamous cell differentiation accompanied by partial and minor interstitial edema. Peripheral palisading cells around tumor nests were also found. The infiltrating depth of tumor ranged from 0. 5~10 mm with the average 4. 12 mm. Cancer embolis were available in 2 cases with no lymph node metastasis. 10 cases followed up for 3~78 months with no recur-rence and metastasis, but 2 cases were lost. Conclusion Adenoid basal cell carcinoma is a rare uterine cervical tumor found in post-menopausal women, which are often accompanied with CIN. This tumor has a favorable prognosis and should be clearly separated from adenoid cystic carcinoma and other tumors which also have infiltrative growth pattern.

Objective Hermaphroditic planarians possess a very important position in the systematic evolutionary history of animal, as they are capacity of complete regeneration. Hence, the research on histological structure of autofluorescence has been carried out to provide a crucial insight into the developmental and regenerative biology. Methods Part of histological structure of planarian (Dugesia japonica) was revealed with HE method, Masson method and Van Gieson method. Their autofluorescence was observed with ultraviolet. There were six planarians in each stained group and the autofluorescence group. Results Epidermis, outer epidermis of pharynx, protonephridium, intestine, the photoreceptor cells and longitudinal nerve cords, all radiated blue autofluorescence. The epithelial dissociation side of copulatory bursa radiated yellow autofluorescence, its middle part radiated blue autofluorescence, its fundus side radiated weakly blue autofluorescence. Testis could hardly give off autofluorescence. Pigment cells of eyepot could not give off autofluorescence. Conclusion The research on configuration and autofluorescence of planarian eye may offer help for the study of origin and evolutionary law on eye of invertebrate.

BACKGROUND: Transplantation of microencapsulated rabbit Schwann cells in the rat spinal cord can relieve inflammatory reaction, promote spinal cord regeneration, but the precise mechanisms remain unclear. OBJECTIVE:To observe basic fibroblast growth factor (bFGF)expression and movements recovery following transplantation of microencapsulated rabbit Schwann cells in rat spinal cord. METHODS: The sciatic nerves taken out from rabbits wore digested with mixed enzyme and were made into Schwann cells suspension. Then we used air-jet method to make Schwann cells microcapsule. Using the same method, empty microcapsule was made. Sprague Dawiey rats were randomly divided into cell group, empty microcapsule group and microcapsule group. Conducted by hemisection injury of spinal cord,the rats in cell group,empty microcapsule group and microcapsule group were implanted with gelatin sponge with 10μL Schwann cells suspension, gelatin sponge with 10 μL empty microcapsule and 10 μL microencapsulated Schwann cells. Normal group was left intact. After operation, we observed hindlimb movements recovery in rats with the Basso, Beattie, and Bresnahan (BBB) scale. Meanwhile,a set of sections were stained immunohistochemically for bFGF expression, another set of sections wore stained for hematoxylin-eosin and Nissal. RESULTS AND CONCLUSION: After spinal cord injury, rat right hindlimb affected paralysis immediately. At 7, 14 and 28 daysfollowing transplantation,motor function in rat hindlimb was significantly recovered, and the BBB scores were significantly higher in microencapsulated schwenn cells than in cell and empty microcapsule group (P < 0.05 or P < 0.01). bFGF positive products were mainly distributed in cytoplasm of the spinal neuron and nucleus of neuroglical cell. The numbers of bFGF positive glial cells mainly appeared surrounding the spinal cord injured site on days 1, 3, 7 and rose to its peak on day 3 and began to appear in neuronal calls on day 14. The number of bFGF positiv cells in microcapsule group was significantly superior to that in cell group and empty microcapsule group. From then on, the bFGF expreSsion was significantly decreased in each group. These indicated that transplantation of microencapsulated Schwann cells can inhibit the immunological rejection after xenotransplantation, suppress inflammatory reaction, improve the expression of bFGF, increase hindlimb movements recovery and spinal cord regeneration after spinal cord injury.

To analyze influenza pathogen spectrum in Yunnan province during 2009-2014 years, and analyze HA and NA genes of influenza A H1N1. Analysis was made on the monitoring date of influenza cases in Yunnan province in recent 6 years, 23 strains of influenza virus of HA and NA gene was sequenced and analyzed by MEGA 5 software to construct phylogenetic tree. 4 times of influenza AH1N1 epidemic peak were monitored from 2009-2014 years in Yunnan Province, as the nucleic acid detection results of influenza A H1N1 accounted for 28.8% of the total. The sequencing result showed that HA and NA gene were divided into 3 groups, one was detected with H275Y mutation strains. Influenza A H1N1 is one of the important subtypes in Yunnan province and their genes have divided into three branches during the period of 2009-2014 years, the vast majority of influenza a H1N1 are still sensitive to neuraminidase inhibitors.
China ; epidemiology ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; metabolism ; Humans ; Influenza A Virus, H1N1 Subtype ; classification ; enzymology ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Molecular Sequence Data ; Mutation ; Neuraminidase ; genetics ; metabolism ; Phylogeny ; Viral Proteins ; genetics ; metabolism

AIM:To investigate the protective effect of losartan(Los) on apoptosis of H9c2 cells induced by isoprenaline(ISO),and to discover its related mechanism.METHODS:H9c2 cells cultured on plastic plates were divided into control,ISO,ISO+Los,ISO+Los+LY294002 and DMSO groups.Cell apoptosis was evaluated by flow cytometery and agarose gel electrophoresis.The mRNA levels of bax,bcl-2 and caspase-9 were detected by RT-PCR and the expressions of phosphorylated and total Akt(p-Akt and t-Akt) were assessed by Western blotting.The cAMP was measured by radioimmunoassay.RESULTS:ISO at concentration of 10 ?mol/L induced apoptosis of H9c2 with an increase in bax/bcl-2,caspase-9 and cAMP.Addition of 10 ?mol/L losartan inhibited apoptosis obviously with a decrease in bax/bcl-2,caspase-9 and cAMP.A significant increase in p-Akt was observed,and its protein level was elevated.LY294002 at concentration of 1 ?mol/L abolished the protective effects of losartan on ISO-induced apoptosis in H9c2 cells.CONCLUSION:ISO might induce H9c2 cell apoptosis through stimulation of ?-adrenergic receptor(?-AR).Los inhibits downstream signaling of ?-AR,and promotes the activation of Akt.Subsequently it might attenuate the apoptosis induced by ?-adrenergic stimulation of ISO.

Objective To evaluate the accuracy and feasibility of time-resolved immunofluorometric assay (TRI FA) for detection of HBsAg based on Abbott automated chemiluminescence immunoassay(CMIA),so as to carry out this project in primary hospitals,and provide reference for individual antiviral strategy and prediction of therapeutic effect.Methods Serum of 157 patients infected with hepatitis B virus were detected with CMIA and TRIFA,specimens with HBsAg titers exceeding the detection limit were firstly diluted,then performed quantitative analysis.HBsAg levels were divided into 4 groups:≤100 IU/mL,101-1 000 IU/mL,1 001-20 000 IU/mL,and ＞ 20 000 IU/mL,quantitative correlation between two methods was analyzed.Results The linear regression equation of two methods was Y=2.323X-896.3,correlation coefficent r=0.943,P＜0.001.CMIA was as a reference,4 groups were divided for analysis,results showed that when detected specimens was at low concentration of HBsAg,TRIFA value was low compared with CMIA method,while detected specimens was at high concentration of HB sAg,CMIA value was high,two reagents had good consistency in the detection of different concentrations of HBsAg(both P＜0.05),when concentration was at 1 001-20 000 IU/mL,consistency was the best.Conclusion The accuracy of two reagents in the quantitative detection of HBsAg is similar,and the best correlation of detection value is 1 000-20 000 IU/mL.TRIFA assay has wide application for its low-cost and easy to be operated,which is especially suitable for primary hospitals.

Objective To investigate the effect of TGFβ1 siRNA on hepatic satellite cells (HSCs) activation, proliferation and extracellular matrix production. Methods The TGFβ1, siRNA plasmid was transfected into HSCs with Lipofectamine 2000. The supernatant and HSCs were collected after incubation for 72h. The expression of TGFβ1, and a-SMA protein in HSCs was detected by. Western blotting. The expression of type Ⅰ and Ⅲ collagen mRNA was detected by RT-PCR. The cell proliferation was assayed by MTT method. Contents of typeⅣ collagen and hyaluronic acid in supernatant were determined by radioimmuno-assay.Results Compared with scrambled control group, the TGFβ1, and a-SMA protein expression,the activity of HSCs proliferation,the expression of typeⅠ and Ⅲ collagen mRNA,and the contents of type Ⅳ collagen and hyaluronic acid in supernatant were reduced in TGFβ1, siRNA group by (79±5)%,(55±4)%, (25±4)% ,(63±6)% ,(57±4)% ,(53±8)% ,(46±8)% ( P<0.01),respectively.Conclusion TGFβ1, siRNA could significantly reduce the expression of TGFβ1,inhibited HSC activation,proliferation and extracellular matrix production.