Objective To analyse the mutation types of the NOTCH3 gene with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy(CADASIL). Methods Including 4 probands of CADASIL,10 cases of CADASIL and 4 normal members from 3 CADASIL families and 100 healthy controls were recruited. Genomic DNA was extracted from white blood cell. The amplicons were analyzed by the denaturing high-performance liquid chromatography (DHPLC) technique. The positive samples which identified by the DHPLC were sequenced to determine the specific mutation or polymorphism, respectively. Results Three heterozygous missense mutations including Cys90Arg, Arg141Cys, Cys134Tyr located in the exon3 and exon4 were found in the 4 probands and 10 cases of CADASIL among the 3 families. 15 polymorphisms were also found. 2 members individual from family 1 and 2 were found to carry the same pathological mutations as in their proband but without clinical symptoms. They were identified as preclinical patients. Conclusions Mutation detection of NOTCH3 is the molecular genetic mechanical for CADASIL. The exon3 and exon4 are possible hot mutation spots in Chinese patients. The mutation of Cys134Tyr in exon4 is a novel mutation which has not been reported previously in China.

The DNA microarray study showed that in STZ-induced diabetic rats the elevatedexpression of 42 genes in artery were depressed markedly after atorvastatin treat ment .This suggeststhat atorvastatin may have the protective effects on the diabetic vascular lesion.

Objective To explore the effect of intensive pedaling training on lower limbs recovery from hemiplegia. Methods 60 hemiplegics after stroke were divided into treatment group (n=30) and control group (n=30). Both groups accepted routine rehabilitation, including physiotherapy, occupational therapy, acupuncture and electrical stimulation, while the treatment group accepted intensive pedaling training in addition. They were evaluated with Berg Balance Scale (BBS), Fugl-Meyer Assessment of lower limb (FMA-L), modified Barthel index (MBI), Functional Ambulation Category (FAC), modified Ashworth scale (MAS) before and after 8 weeks of treatment. Results The scores of BBS, FMA-L, MBI, FAC, and MAS improved in both groups after treatment (P<0.01), and improved more in the treatment group than in the control group (P<0.05). Conclusion Intensive pedaling training can further improve the lower limbs function of stroke patients with hemiplegia

Objective To check the maneuverability of the nursing safety management indicators for skilled nursing facilities.Methods A hospital based skilled nursing facility in Chongqing was chosen as the main study target while a Red Cross nursing hospital in Shanghai which operated earlier was also included as a comparison object.Combination of files reference,on-the-scene observation,manager interviews,sampling survey and institution's self-assessment,scores of indicators were calculated,and the total scores for both institutions were calculated as well.Results The institution from Chongqing marked 81 points while the other one got 86 points.Conclusions The indicator system had good maneuverability and the weight of indexes was consistent with the practical requirement of safety management.This indicator system could provide reference for standardize skilled nursing facilities' management,but still needed further revision and consummation.

Objective To evaluate the effects of ELISPOT (enzyme-link immunospot) test using different samples in diagnosing tuberculous pleurisy. Methods Using T-Spot-TB kit to detect interferon-γlevel in pleural effusion and periph?eral blood from 164 patients with tuberculous pleural effusion and 102 patients without tuberculous pleural effusion. Number of spot forming cells (SFCs) as well as the specificity and sensitivity of the tests were compared between these two methods (ELISPOT using leural effusion or peripheral blood). Results The area under the ROC curve was 0.947 in pleural effusion Elispot test while it was 0.905 in peripheral blood Elispot test. The sensitivity of pleural effusion ELISPOT test in diagnosis of tuberculous pleurisy (95.1%) was significantly higher than that of peripheral blood ELISPOT test (89.0%). What’s more, the specificity of pleural effusion ELISPOT test in diagnosis of tuberculous pleurisy (90.2%) was higher than that in diagno?sis of peripheral blood ELISPOT test (88.2%). Conclusion The pleural effusion ELISPOT test is more valuable than periph?eral blood ELISPOT in the diagnosis of tuberculous pleuritis.

ObjectiveTo explore the effect of intermedin pretreatment on repair and regeneration process of renal ischemia reperfusion (IR) injury.Methods Male Wistar rats were randomly divided into four groups: sham group,IR group,empty plasmid group and IMD group.One week after removing the right kidney,eukaryotic expression vector encoding rat IMD gene was transfected into the left kidney using an ultrasound-microbubble-mediated system,and transfection effect was detected by Western blotting and RT-PCR.After successful transfection,renal IR injury model was induced by clamping left renal arteries for 45 minutes followed by reperfusion.BUN and Scr were tested.Renal pathology was observed by PAS and HE staining.The proliferation of tubular epithelial cells was detected by SABC immunohistochemical method.Results (1)The expression of IMD protein and mRNA in IMD group was significantly up-regulated compared to empty plasmid group(P＜0.05),reaching the peak at day 7,and the difference was not significant with that at day 4.(2)Compared with sham group,BUN and Scr were obviously increased at day 1and 2 in IR group(P＜0.05).However,compared with IR group,BUN and Scr in IMD group decreased obviously(P＜0.05).Differences between IR group and empty plasmid group were not significant(P＞0.05).(3)Kidney tubules in IR group,IMD group and empty plasmid group were injuryed,but the injury in IMD group was less serious compared with that in IR group and empty plasmid group.The injury in three groups was the most severe at day 2 after reperfusion.(4)The number of PCNA positive cells evidently increased at day 7 in IR group and empty plasmid group,but the number of PCNA positive cell evidently increased at day 3 in IMD group.Compared with IR group 1 to 4 days after perfusion,PCNA positive cells in IMD group increased obviously (P＜0.05).Compared with IR group day 7,PCNA expression in IMD group 7 d was obviously decreased(P＜ 0.05).Conclusion IMD pretreatment can induce tubular epithelial cells proliferative and accelerate the repair and regeneration progress in IR injury kidneys.

Objective To investigate the clinical application value of genechip detection system in the mycobacterial species iden-tification and drug resistance analysis .Methods The specimens of sputum ,punctured pus ,pleural and abdominal ascites ,cerebro-spinal fluid and so on were performed the Mycobacterium DNA detection by using the gene chip technique .Then Mycobacterium tuberculosis positive samples were further performed the drug-resistant analysis .Meanwhile the Ziehl-Neelsen acid-fast staining was adopted to detect the sample .The positive rates were compared between the two groups .And TB-IGRA was used to examine the tubercle bacillus infection in partial patients .Results In 4402 samples ,137 cases (3 .36% ) of M ycobacterium tuberculosis (MTB) and 11 cases(7 .4% ) of non-tuberculosis mycobacterium(NTM) were detected .Puncture solution ,bronchoalveolar lavage fluid and tissue specimen had higher positive rate .In the 137 positive M TB samples by rifampicin-resistant gene rpoB ,isoniazide-re-sistant gene katG and inhA detection ,22 cases of resistance gene mutations were detected ;the positive rate of genechip for detecting sputum ,cerebrospinal fluid ,hydrothorax and ascites was higher than that of acid-fast staining .TB-IGRA detection had higher pos-itive rate of TB infection than genechip .Conclusion The genechip detection system can directly conduct Mycobacterium identifica-tion and drug resistance analysis ,which is especially suitable for sputum ,cerebrospinal fluid ,hydrothorax and ascites samples ,and which is simple and rapid with higher sensitivity and good specificity .

Objective To evaluate the clinical value of narrow band imaging(NBI)for diagnosis of malignant and premalignant gastric lesions.Methods The gastric lesions,pits and microvascularity were observed using conventional endoscopy followed by narrow band imaging(NBI)and chromoendoscopy(0.2%indigo carmine)as well as magnifying endoscopy(×80)in 217 patients.The quality of images obtained by different endoscopies was evaluated and compared to pathologic interpretations. Results Of 217 patients,non-atrophic gastritis was found in 85 cases,chronic atrophic gastritis in 38 cases,mild dysplasia in 19 cases,moderate dysplasia in 9 cases,severe dysplasia in 4 cases,early gastric cancer in 5 cases,advanced gastric cancer in 20 cases and intestinal metaplasia in 91 cases.The NBI endoscopy was superior to conventional endoscopy and chromoendoscopy in finding gastric lesions(P=0.000).The gastric microvascularity was more clearly seen on images obtained by NBI combined with magnifying endoscopy in comparison with conventional endoscopy and chromoendoscopy(P=0.000).There were six patterns in description of gastric pits with NBI endoscopy.Type Ⅲ,Ⅳ or Ⅴ1 was usually found in chronic atrophic gastritis,type Ⅲ,Ⅳ,Ⅴ1 or Ⅴ2 in intestinal metaplasia,type V1 or Ⅳ in dysplasia and type Ⅵ in suspected malignant lesion.Conclusions NBl with magnifying endoscopy is helpful in improving the biopsy accuracy of malignant and dysplastic lesions and in detecting early gastric cancer.

ObjectiveTo evaluate the effectiveness and safety of high frequency electronic associating stents insertion and subsequent radiotherapy guided by fiberoptic bronchoscope in treating tracheobronchial stenosis caused by lung and esophagus carcinoma.MethodsFifty-two patients which were ascertained with serious stenosis in trachea or bronchi by fiberoptic bronchoscope were subjected to the study.Fiberoptic bronchoscope was used to investigate the location,extent,blood supply and the degree of tracheobronchial stenosis before the study.After the focus in the airways was cleared up with high frequency electronic,electrocoagulation and snare under the guide of the euthyphoria of fiberoptic bronchoscope,Ni-Ti memory alloy stents were placed into the stenosis airways.The 52 patients were divided into two groups with 26 cases each:the stent group and the stent associating radiotherapy group(radiotherapy group).The later group accepted subsequent radiotherapy after the therapy.The following indexes of the two groups were investigated:short-time clinical effect,dyspnea index class,rate and average time of airway restenosis,life span and survival rate.ResultsAfter therapy,the 52 patients had obvious improvements in dyspnea and the diameter of the stenosis airways.The short-time total effective rate was 100.0％ (52/52),but there was not significant difference between the two groups(P＞ 0.05 ).After therapy,there was significant difference in two groups in dyspnea index class(P ＜ 0.01 ).Although there was not significant difference in the rate of airway restenosis between the two groups(P＞0.05),the average time of airway restenosis in the radiotherapy group was obviously longer than that in the stent group(P＜ 0.01 ).The survival rate of the radiotherapy group in the 6th and 9th month after therapy was higher than that in the stent group (P ＜ 0.05),but there was no significantdifferenee in the 3rd and 12th month (P ＞0.05).ConclusionHigh frequency electronic associating stents insertion and subsequent radiotherapy guided by fiberoptic bronchoscope is an effective and safe treatment for the patients suffering tracheobronchial stenosis caused by lung and esophagus carcinoma.

Objective To increase the killing effect of aminolevulinic acid (ALA)-based photodynamic therapy (PDT) on a human skin squamous cell carcinoma cell line A431 by poly lactic-co-glycolic acid nanoparticles (PLGA NPs).Methods ALA-loaded PLGA NPs (ALA PLGA NPs) were prepared using a modified double emulsion solvent evaporation method,and characterized in terms of size,encapsulation efficiency,loading capacity and morphology.Transmission electron microscopy was carried out to observe the morphology of A431 cells after uptake of ALA PLGA NPs.To optimize incubation time,multi-mode microplate reader was used to describe the fluorescence kinetics of protoporphyrin Ⅸ generated by A431 cells during 24 hours of incubation with 0.1 mmol/L ALA,1 mmol/L ALA,2.7 g/L ALA PLGA NPs containing about 0.1 mmol/L ALA,and PLGA NPs without ALA separately.Some A431 cells were divided into 10 groups:control group receiving neither treatment nor irradiation,0.1 and 1 mmol/L ALA dark/PDT group incubated 0.1 and 1 mmol/L ALA respectively,ALA PLGA NP dark/PDT group incubated with ALA PLGA NPs of 2.7 g/L,PLGA NP dark/PDT group incubated with PLGA NPs of 2.7 g/L,simple irradiation group irradiated with a He-Ne laser (wavelength:635 nm,power density:8.6 mW/cm2,energy density:8 J/cm2) only.The dark groups were kept in darkness strictly by wrapping in aluminum foil,and PDT groups were irradiated using a He-Ne laser.After another 24 hours of culture following irradiation,methyl thiazolyl tetrazolium (MTF) assay was conducted to estimate the survival rate of cells.To study the effect of ALA PLGA NPs on cell apoptosis,some A431 cells were divided into three groups:control group receiving neither treatment nor irradiation,ALA-PDT group and ALA PLGA NP PDT group receiving PDT after incubation with ALA and ALA PLGA NPs respectively.Flow cytometry was performed to detect the apoptosis of A431 cells at 12 and 24 hours,separately,after the photodynamic therapy.Data were statistically analyzed using SPSS 13.0 software package by means of a t test.Results The prepared ALA PLGA NPs were spherical with a mean particle size of (65.6 ± 26) nm,encapsulation efficiency of (65.8 ± 7.2) ％,and drug loading capacity of (0.62 ± 0.27)％.ALA PLGA NPs could be uptaken by A431 cells and gathered in the cytoplasm.The PpIX fluorescence kinetic study showed that the fluorescence intensity increased with time within 24 hours in A431 cells incubated with ALA or ALA PLGA NPs.After 6 and 24 hours of incubation,the A431 cells incubated with 2.7 g/L ALA PLGA NPs showed a significant increase in the protoporphyrin Ⅸ fluorescence intensity compared with those incubated with 0.1 mmol/L ALA (both P ＜ 0.01).Further more,the survival rate of A431 cells was statistically lower in the ALA PLGA NP PDT group than in the 0.1 mmol/L ALA PDT group at 6 and 24 hours (t =35.685,5.262,respectively,both P ＜ 0.01).Elevated apoptosis rate was observed in the ALA PLGA NP PDT group compared with the ALA PDT group at 12 ((13.10 ± 0.50)％ vs.(4.90 ± 0.13)％,t =9.074,P＜ 0.01) and 24 ((30.17 ± 1.02)％ vs.(11.6 ± 0.59)％,t =9.095,P ＜ 0.01) hours.Conclusions ALA PLGA NPs can promote the formation of protoporphyrin Ⅸ,strengthen the killing effect of ALA-PDT on A431 cells in vitro,and enhance the apoptosis induced by ALA-PDT in tumor cells.