AIM: To construct recombinant adenoviral vector carrying the LEDGFp52 gene by homologous recombination in bacteria and to detect its expression in vitro.METHODS: The LEDGFp52 gene was cloned to adenoviral shuttle plasmid pAdTrack-CMV. Then, the resultant pAdTrack-CMV-LEDGFp52 was cotransfected into BJ5 183 bacteria with the adenoviral backbone plasmid pAdeasy-1. The adenoviral plasmid carrying LEDGFp52 was generated with homologous recombination in bacteria, and the adenoviruses were produced in 293 cells. These 293 cells were then infected with adenoviruses, and the expression of LEDGFp52 was detected by CPE (cytopathic effect) and western blot.RESULTS: The titer of Ad-LEDGFp52 adenoviruses was up to 5×1012 pfu/L after proliferation in 293 cells. LEDGFp52 was expressed efficiently in 293 cells after infection.CONCLUSION: The recombinant adenoviruses vector expressing LEDGFp52 was constructed successfully and can be used in further gene transfection experiments.

AIM: To construct and identify LEDGFp52 eukaryotic expression vector for RNA interference.METHODS: Recombinants were designed and established by targeting gene LEDGFp52 and plasmid pGensil-1 based on LEDGFp52 cDNA sequences of Genomes. Two pairs of oligonucleotides were synthesized according to the Tuschl principle and inserted into plasmid pGenSil-I to generate siRNA eukaryotic expression vector. DH5α strains were transformed, plasmids were extracted, and recombinant vectors were identified by the restriction map and the sequence analysis. The cultured cells were transfected by the recombinant plasmid (pGensil-1-RNA. LEDGFp52-1). At 48 hours after transfection, the whole cell protein was extracted, and the protein level was detected using Western blotting with mouse anti-human LEDGFp52 monoclonal antibody.RESULTS: Recombinant plasmids completely concord with the designs by the restriction map and the sequence analysis,the proteinlevel of LEDGFp52 was down regulated at 48hours after transfecting pGensil-1- LEDGFp52-1 expression vector into HeLa cells, the recombinant eukaryotic expression vectors were successfully constructed.CONCLUSION: siRNA recombinant can be successfully constructed by RNAi technique to inhibit the expression of LEDGFp52.

AIM: To construct the prokaryotic expression vector of rhLEDGFp52 gene,to obtain the rhLEDGF p52 protein.METHODS: rhLEDGFp52 gene was constructed into a prokaryotic expression vector pET30a (+) by recombinant DNA techniques and was identified by enzymatic digestion and sequence analysis. rhLEDGFp52 protein was induced expression by IPTG in E.coli BL21 (DE3), it was tested by Western blot and was purified by Ni-NTA His. Bind. Resin.RESULTS: We Successfully constructed the prokaryotic expression vector of rhLEDGFp52 gene and obtained its expression in E.coli BL21 (DE3),it was expressed in a soluble form and detected up to 34.63% of the total bacterial protein expressed in E.coli BL21 (DE3).Western blot analysis demonstrated that rhLEDGFp52 protein could spicifically integrate with LEDGF-ab. After purified by Ni-NTA His. Bind. Resin, the ultimate concentration of purified rhLEDGFp52 protein was 520μ g/ml and its purity was 87.93%.CONCLUSIONS: rhLEDGF p52 protein was obtained that provides an experimental basis for the further study of the biological function of rhLEDGFp52 protein.

ObjectiveUsing large sample clinical study to reevaluate the safety and efficacy of recombinant human growth hormone(ANSOMONE) in treating pediatric growth hormone deficiency(GHD).Methods1012 cases were administered with ANSOMONE at a dose of 0.1 IU · kg-1 · d -1 in bedtime for 6 months.The alteration of mean height,growth rate,height standard deviation scores,bone age,and anti-GH antibody and occurrence of adverse effects were observed.ResultsThe growth rate was promoted from ( 2.50 ± 0.85 ) cm/year to ( 13.44 ± 4.27 ) cm/year,the net growth rate was( 10.94 ±3.69)cm/year.There was significant difference between the pre-treatment and post-treatment patients ( P ＜ 0.01 ).The post-treatment anti-GH antibody positive rate was 13.34％,which had no obvious effect on the efficacy.Few patients exert transient inflammation reactions,which disappeared with any treatment.ConclusionANSOMONE had significant efficacy,and could promote the height increase obviously.It had good safety without any obvious side effect and early-maturing effect.

OBJECTIVE: To understand the assessment on the criminal responsibility of drug-induced mental disorders and judicial experts' opinions. METHODS: The judicial experts from institutes of forensic psychiatry in Shanghai were selected. They were asked to finish a self-made questionnaire of assessment on the criminal responsibility of drug-induced mental disorders by letters and visits. RESULTS: Most of experts knew the special regulation, "not suitable for evaluation" towards the criminal responsibility of drug-induced mental disorders of the guideline promulgated by Ministry of Justice. Before and after the guideline was issued, no expert made a no-responsibility opinion in such cases. After the guideline was issued, some experts made a full-responsibility or limited-responsibility opinion in such cases. There was a little disagreement among the experts in the case that the crime was unrelated with mental symptoms or the criminals used drugs even though he knew it could induced insanity. But there were still many obvious disagreements among experts in the case that crime was related to such symptoms and person was no ability to debate. Most experts agreed to settle the disagreements with improved legislative perfection. CONCLUSION Most experts are not strictly complying with the assessment guidelines during their practice, and there is still an obvious disagreement towards the criminal responsibility of drug-induced mental disorders.
China ; Crime/psychology* ; Criminals/psychology* ; Data Collection ; Expert Testimony ; Forensic Psychiatry ; Humans ; Liability, Legal ; Male ; Mental Competency ; Mental Disorders/psychology* ; Psychotic Disorders ; Surveys and Questionnaires

This paper investigates the main problems existing in research-type key lab construction in higher schools.Based on the investigation,countermeasures are proposed also.

Objective To develop and evaluate three types of biodegradable intravascular stent (BIS)with biodegradable materials of poly-L-acid(PLLA)or Poly](1-lactide-co-glycol ide)(PLLGA). Methods Forty-five spiral BISs(15 BISs for each type)were prepared with PLLA(153 kDa,type A), PLLGA(141 kDa,type B),and PLLGA(contains paclitaxel,141 kDa,type C).The physical and mechanical properties of the BIS were tested,including radial force,overlay rate,longitudinal shrinkage rate,and rate of expansion.Results The radial force of three types BIS was 15.7%/0.006 Mpa(type A), 16.3%/0.006 Mpa(type B),and 16.4%/0.006 Mpa(type C).The radial force of type A was greater than that of type B and C(P

Objective To investigate the effect of long-term high-fat diet on insulin resistance and the morphology and function of islets in rats and the relationship between them.Methods Thirty normal male Wistar rats (8 weeks old) were divided into two groups and fed either with normal chow (NC,n=15),or high-caloric and high-fat diet (HF,n=15).Insulin resistance was assessed by euglycemic hyperinsulinemic clamp technique. The insulin secretory function of islets was evaluated by intravenous insulin releasing test.Morphological and quantitative analysis of pancreatic tissues was performed by double-label insulin and glucagon immunohistochemistry.Proinsulin mRNA was detected by RT-PCR.Results The glucose infusion rate (GIR) in HF rats was significandy lower than that in NC rats [(5.83?0.79)mg?kg~(-1)?min~(-1) vs (7.60?1.29)mg?kg~(-1)?min~(-1),P＜0.05].Immunohistochemistry showed that HF rats had larger islet size [(15168?1327)?m~2 vs (6264?1840)?m~2,P＜0.01] and significantly reduced insulin relative concentration of?cells[(-5.15?0.03) vs (-4.81?0.17),P＜0.01],as compared with NC rats.The islet relative?cell volume was decreased signifieandy (P＜0.01),whereas the relative?cell volume was increased (P＜0.01).So the ratio of?to?were lower in HF [(4.68?1.01) vs (11.84?3.82),P＜0.05].The peak of insulin secretion in intravenous insulin releasing test in HF was at 10 min,whereas that in NC rats was at 5 min.AUC (area under curve) 10-60 rain of insulin in HF was higher than that in NC rats [(152.51?34.53)mIU?L~(-1)?min~(-1) vs (86.40?21.21) mIU?L~(-1)?min~(-1),P＜0.01].There was no difference in proinsulin mRNA levels between two groups. Conclusion Long-term high-caloric and high-fat diet results in early impairment of islet morphology and function, as well as significant insulin resistance,suggesting that the compensation ability of islets has already been impaired in the early course of type 2 diabetes mellitus.

OBJECTIVETo investigate the expression of cysteine-rich protein 61 (Cyr61) and NF-kappaB in gastric carcinoma and its correlation with the clinicopathologic features and survival time.

METHODSRT-PCR was used to validate and detect expression of Cyr61 mRNA in 53 gastric carcinoma specimens and 11 non-tumor gastric mucosa samples. Cyr61 and NF-kappaB protein levels expressed were detected using immunohistochemistry in 99 gastric carcinoma specimens and 25 non-tumor gastric mucosa samples.

RESULTSRT-PCR demonstrated that expression of Cyr61 mRNA was higher in the primary carcinoma (84.6%, 22/26) and the metastatic foci (88.9%, 24/27) than in the non-tumor control samples (5/11; P < 0.05, respectively). Cyr61 gene mRNA expression levels were (2.76 +/- 5.50) x 10(-5), (14.61 +/- 20.64) x 10(-5), and (18.46 +/- 26.38) x 10(-5) by 2(-DeltaCt) in the control mucosa samples, primary carcinomas and metastatic tissues respectively. The level was higher in the primary carcinomas and metastatic tissues than that of the non-tumor gastric mucosa (P < 0.05, respectively); however, there was no significant difference between the metastatic tissues and the primary carcinomas (P > 0.05). Immunohistochemistry revealed that of the 99 cases, there was a high expression of Cyr61 and NF-kappaB protein, 56.6% (56/99) and 55.6% (55/99) respectively. There was correlation of Cyr61 and NF-kappaB protein expressions with the depth of tumor and vascular invasion, as well as the development of lymph node metastasis, and TNM staging (P < 0.05, respectively), besides, the expression of NF-kappaB also correlated with the tumor diameter (P < 0.05). Cyr61 expression was positively correlated with NF-kappaB expression in gastric carcinoma (P < 0.05); the mean survival time in cases with a high expression level of Cyr61 and NF-kappaB protein was significantly shorter than those with a low expression level (P < 0.05).

CONCLUSIONSExpression of Cyr61 and NF-kappaB closely correlated with invasion and metastasis of gastric carcinoma. They may be considered as the biologic behavior indicators for gastric carcinoma.

Carcinoma ; genetics ; metabolism ; Cysteine-Rich Protein 61 ; genetics ; metabolism ; Female ; Gastric Mucosa ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; diagnosis ; genetics ; Male ; NF-kappa B ; genetics ; metabolism ; Neoplasm Staging ; Prognosis ; Statistics as Topic ; Stomach Neoplasms ; diagnosis ; genetics ; metabolism ; Treatment Outcome

Humans ; Nasal Obstruction ; surgery ; Treatment Outcome ; Turbinates ; surgery