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.

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.

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.

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 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.

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

Objective The use of extracorporeal membrane oxygenation (ECMO) as a treatment for the failure of cardiopulmonary function after cardiac surgery is increasing and has been reported to be 3% to 5% in the cases with congenital heart disease. We reviewed our experience with ECMO in children who received heart surgery for congenital heart disease and complicated with severe heart failure postoperatively. Methods Eight patients received ECMO, seven was due to the failure to wean from bypass and one had fulminant myocarditis. Import membrane oxygenator,veno-arterial mode ECMO and right atriumascending aortic cannulation were used in 7 cases and peripheral cannulation via femoral veno-artery route was used in 1 case.Supportive intervention persisted from 65 to 498 hours, with flow rate maintained at 80 to 120 ml per minute per kilogram body weight. Results Five patients died, with a mortality of 62.5%, and 3 cases discharged, with a survival rate of 38%. Bleeding occurred in 5 cases, thrombosis occurred in 2 cases, hemolysis was identified in 1 case and DIC was observed in 1 case.One case had liver failure and 2 cases had malnutrition. Oxygenator plasma leakage occurred in 2 cases. Mean arterial blood pressure increased significantly after the establishment of ECMO as compared with that before the procedure [( 60.2 ± 7.8 )mmHg vs. (48. 1 ± 5.2 ) mmHg, P≤0.05]. The arterial concentration of lactate decreased significantly, from (5. 1 ± 0. 8 )mmol per liter before ECMO to ( 3.6 ±0. 5 )mmol per liter after ECMO, P ＜0.05. Conclusion For patients who survived the congenital heart surgery and no residual anatomic deformity, ECMO can be used as early as possible as a treatment for severe heart failure which resulted from coexistent of left and right ventricular and pulmonary insufficiency. An overall mortality may be decreased by ECMO technique as it plays a substitution role for gas exchange in the lung. As a result, the concentration of oxygen and the airway pressure used during ventilation, and the resultant lung injury can be reduced. Appropriate strategies involve transfusion of fresh platelet and packed red blood cells, replacement of frozen plasma and blood products, as well as rational use of vasoactive drugs and heparin, and maintaining a stable internal environment. Following strategies are also recommended: using continuous arterio-venous hemofiltration and durable heparin-coated membrne oxygenator, reducing hemorrhagic complications, monitoring pressure on both side of the film, identifying plasma leakage carefully and reducing the mechanical complications.

OBJECTIVE: To summarize the effects of glial cell line-derived neurotrophic factor (GDNF) on ischemia damage to nerve tissue and discuss the possibility of GDNF in repair of spinal cord injury based on the development of microencapsulation technology.DATA SOURCES: A search of Medline from January 1996 to October 2000 was performed for the English articles related to GDNF, ischemia damage to nerve tissue, spinal cord injury and microencapsulation technology by using the key words "glial cell line-derived neurotrophic factor, ischemia damage to nerve tissue, spinal cord injury". Meanwhile, we retrieved Wangfang database for search of the related articles in Chinese by using the same keywords in Chinese.STUDY SELECTION: Articles including intervention group and control group were selected after first review, and those which were significantly non-randomized researches were excluded. Then, the full-texts of the enrolled articles were retrieved. Inclusion criteria: ①randomized controlled study; ②the experiment/clinical research including horizontal control group. Exclusion criteria: duplicated researches.DATA EXTRACTION: Totally 300 articles were selected but only 15 were in coincidence with conclusion criteria. 285 articles were excluded, 264 of them were duplicated and non-randomized researches, and 21 were review articles.DATA SYNTHESIS: GDNF can provide nutrition to dopamine nerve cell in rat's middle brain, so as to decrease dopamine nerve cell's death. Also GDNF can protect dopamine nerve cell in cerebral infarction rats from ischemic injury, inhibit the produce of nitrogen monoxide and reperfusion injury after ischemia. GDNF is an effective protective factor against ischemia damage. Microencapsulation technology has a bright future in treating endocrinopathic neural diseases, and GDNF can play a great role in the development of microencapsulation technology.CONCLUSION: GDNF is a protective factor against ischemia damage to nerve tissue, which can be enhanced by microencapsulation technology.There is a bright future for the research on GDNF in the clinical repair of spinal cord injury.

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.