Objective In cartilage tissue engineering,it was a difficult problem to keep chongdro-cytes phenotype for long time.The aim was made to investigate the dedifferentiation of human articular chon-drocytes,and to verify the inhibition to dedifferentiation by transfection with recombinant pcDNA3.1(+)/TGF-? 2 in vitro.Methods Free chondrocytes were isolated from healthy articular cartilage harvested from6adult patients underwent operation because of other condition.The recombinant pcDNA3.1(+)/TGF-? 2 con-structed in advance was transferred by liposome mediated transfection to monolayer cultured articular chon-drocytes.The first,sixth and ninth generation chondrocytes being transfected and non-transfected were as-sayed by histological examination of HE and Safranine-O staining to observe their morphological changes.RT-PCR,ELISA,immunohistochemistry analysis and prime hybridization were performed to these cells to i-dentify the expression and biosynthesis of TGF-? 2 and cartilage-associated genes and proteins.Results The non-transfected articular chondrocytes in monolayer culture demonstrated the tendency of dedifferentia-tion,and lost the characteristics of their phenotype progressively in subculture.The expression of TGF-? 2 ,collagen typeⅡand proteoglycan in non-transfected cells were decreased,but expression of collagen typeⅠincreased gradually with time going on.On the contrary,as for the transfected chondrocytes,the trans-ferred gene was expressed in cells of all examined generations,and the expression levels were higher than that of non-transfected cell of same generations.The transfected chondrocytes maintained their phenotype and characteristics of articular cartilage.They expressed and produced collagen typeⅡand proteoglycan in a higher level,and collagen typeⅠin a lower level than that of non-transfected cells.Conclusion Human articular chondrocytes in monolayer culture have the tendency of dedifferentiation.The results suggested that chondrocytes tranfected by pcDNA3.1(+)/TGF-? 2 could produce endogenous TGF-? 2 .Transfection of pcD-NA3.1(+)/TGF-? 2 is able to provide transient and long lasting expression of TGF-? 2 in chondrocytes,and may be a feasible method to inhibit dedifferentiation of articular chondrocytes in vitro.

Objective To study CT features of liver abscesses caused by the fasciola hepatica infection , and discuss its pathologic basis.Methods CT images of 15 Patients were collected. All patients underwent both unenhanced and biphasic enhanced CT scanning, then its CT performances were analyzed. Results round and nodular lesions were observed in 15 cases, branching and stripping lesions like dilated bile duct in 9 cases. The density of lesions was inhomogeneous, and the lesions were multifocal and multiform. The liver abscesses caused by the fasciola hepatica infection had no “rim sign” or “target” sign, Liver abscesses were less than 3.0 cm in diameter, and the dilation of the bile duct were not observed. Conclusion Liver absessed caused by the fasciola hepatica infection have characteristic CT features. Combined with clinical examination and laboratory test, the reliability of diagnosis will be considerably increased.

Objective: To investigate the public opinion on pharmaceutical service and explore the implementation of pharmaceutical service fee and the prospect of online pharmaceutical service in our country.Methods: Questionnaires which related to pharmaceutical service fee and online pharmaceutical service were distributed to patients and their relatives or friends, and then recycled.The data including the recycling of questionnaires, information of respondents and answers to the questions were statistically analyzed.Results: Totally 63.8% respondents considered pharmaceutical service fee was reasonable and the fee should be borne by the government finance and health care insurance.Totally 85.5% respondents thought launching online pharmaceutical service was necessary, which included online retails of medicine, consultation, medication guidance, decoction of Chinese traditional medicines, medicine delivery and so on, especially pharmacy consultation.Conclusion: Outpatients have high demands of pharmaceutical service and most of them accept online pharmaceutical service with different understandings.Pharmaceutical service fee is an inevitable trend in the development of pharmaceutical industry.More publicity and guidance should be given to improve people's acceptance of pharmaceutical service and relevant cost.Relevant departments should take full advantages of Internet and provide diversified pharmaceutical service.

BACKGROUND: Some studies have presented that vascular endothelial growth factor and its receptor system may take part in onset and development of diabetic nephropathy (DN).OBJECTIVE: To further verify the interventional effects of irbesartan on vascular endothelial growth factor (VEGF) and its Flk-1 receptor expressions in kidney of diabetes mellitus (DM) rat, and the possible mechanism of irbesartan.DESIGN: Randomized control animal study.SETTING: West China Hospital of Sichuan University.MATERIALS: Eighteen male closed colony SD rats weighing 150-200 g were cared in standardization.METHODS: This study was performed at Laboratory of West China Hospital of Sichuan University from August 2006 to April 2007. All rats were randomly divided into a DN group, an irbesartan group and a normal control group, with 6 rats in each group. 10 g/L streptozotocin (55 mg/kg) was intraperitoneally injected to establish models; rats in the control group were administrated with the same dosage of citric acid buffer solution; rats in the irbesartan group were administrated with 3 mg/(kg·d) irbesartan after model establishment. VEGF and FIk-1 expressions were detected using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry technique; while, urine protein level, area and volume of renal glomerulus were detected, and correlations of data were analyzed.MAIN OUTCOME MEASURES: ①Renal pathological indicators, urine protein level, area and volume of renal glomerulus; ②VEGF and Flk-1 expressions;③renal immunohistochemical examination;④ correlation analysis.RESULTS:① Renal pathological examination by HE staining indicated that renal glomerulus was remarkably enlarged in the DN group; mesangial matrix was increased; mesangial cells were also increased; renal tubule was expanded. The lesions in the irbesartan group were milder compared to the DN group. Urine protein level in the DN group was significantly higher compared to the control group (P ＜ 0.01); renal weight/body mass, area and volume of renal glomerulus were significantly higher compared to the control group (P ＜ 0.01); urine protein level, renal weight/body mass, area and volume of renal glomerulus in the irbesartan group were significantly lower compared to the DN group (P ＜ 0.01). ② By the 16th week,VEGF and Flk-1 expressions in the DN group were significantly up-regulated compared to control group (P ＜ 0.05); while,VEGF and Flk-1 expressions in the irbesartan group were also up-regulated compared to the control group by the 16th week (P ＜ 0.05) but down-regulated compared to the DN group (P ＜ 0.05). ③ VEGF staining in the DN group was darker compared to control group (P ＜ 0.01), while the staining in the irbesartan group was also darker compared to the control group (P ＜0.01), but the staining in the irbesartan group was lighter compared to the DN group (P ＜ 0.01). Flk-1 staining was similar to the VEGF. ④ VEGF and Flk-1 were positively correlated, with urine protein level, area and volume of renal glomerulus (P ＜0.05).CONCLUSION: VEGF and its Flk-1 receptor play important roles in DN pathogenesis. Over expressions may cause renal injury, but irbesartan (angiotensin Ⅱ receptor antagonist) has the protective effects on kidney through inhibiting abnormal expression of VEGF and Flk-1.

Clinical laboratory biological safety is one of whole society safety. This paper introduced briefly the current situation of clinical medical laboratory biosafty in the hospital. and set forth common biological hazards specifically for whose characteristics. Combining the biosafety administration measures from abroad, the issue of laboratory biological safety administration was considered, and put forward some suggestions according to related law and regulation of national laboratory safety administration in order to strengthen clinical laboratory biosafety administration.

OBJECTIVE:To provide reference for the better development of intemet pharmaceutical service of hospital pharmacists.METHODS:The questionnaire investigation was carried out on the cognition of pharmacists to intemet pharmaceutical service in 9 third grade class A hospitals.The survey data were analyzed statistically.RESULTS:A total of 600 questionnaires were distributed and 527 questionnaires were collected with recovery rate of 87.8％.Among them,there were 499 valid questionnaires with effective rate of 94.7％.The most selected internet pharmaceutical service contents were medication consultation (94.4％) and medication education (91.6％).93.0％ considered that intemet pharmaceutical service were worthy or very worthy of promotion.The most concerned issues about intemet pharmaceutical service were technical problems (58.5％) and legal liability (55.9％).57.1％ of the surveyed pharmacists considered that they were totally or possibly capable of performing interact pharmaceutical care;37.9％ didn' t get in touch with intemet pharmaceutical care and were not clear about it.The most important qualifications of pharmacists who provided intemet pharmaceutical service were rank of pharmacist's title (67.9％) and working experience (67.7％).It was considered that the most effective measures to ensure the quality of intemet pharmaceutical care were regular pharmaceutical service training (74.3％) and proportionally sampling the quality of pharmaceutical service consultation (67.5％).CONCLUSIONS:Internet pharmaceutical service may become a new direction of the development of pharmaceutical service under new medical reform.The development of intemet pharmaceutical service requires perfect laws and regulations,and pharmacist team with high professional level.

AIMTo detect the hepatotoxic pyrrolizidine alkaloids (HPA) in the genus Ligularia Cass..

METHODSThe alkaloid extracts of Ligularia plant materials were detected and analyzed by the method of combination of TLC, and LC/MSn.

RESULTSAmong 22 species of Ligularia Cass., HPA were detected in 18 species with LC/MSn, and no HPA was detected in the remaining 4 species.

CONCLUSIONHPA was first detected with LC/MSn in L. tongelensis and other 15 species of Ligularia Cass.; HPA from these plants should be isolated, separated and identified and it is necessary to study the activities and toxicities of the HPA. The types and kinds of HPA from different species and sources are different, they should be detected separately.

Asteraceae ; chemistry ; classification ; Chromatography, Thin Layer ; Molecular Structure ; Plants, Medicinal ; chemistry ; Pyrrolizidine Alkaloids ; analysis ; chemistry ; Species Specificity ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo explore mechanisms of acupuncture for reducing of body weight.

METHODSThe rat obesity model was developed by subcutaneous injection of sodium glutamate solution, and the experimental rats were divided into a blank control group, a model group, an electroacupuncture group and a sibutramine group; blood TC, TG, HDL-C, LDL-C contents and lipoprotein lipase (LPL) activity, and serum leptin and insulin levels were determined.

RESULTSThe body weight and Lee's index, and TG, TC and LDL-C contents in the electroacupuncture group decreased significantly as compared with those in the model group (all P<0.01), the actions in the electroacupuncture group being superior to those in the sibutramine group; the HDL-C contents in both the electroacupuncture and the sibutramine groups were higher than that of the model group (P<0.01) with no significant difference between the electroacupuncture group and the sibutramine group; LPL activity in the electroacupuncture group increased significantly as compared with that in the model group (P<0.01); serum leptin and insulin levels in both the electroacupuncture group and the sibutramine group were lower than that in the model group (P<0.05, P<0.01), with no significant difference between the electroacupuncture group and the sibutramine group in the effect on insulin content, and the decrease of leptin level in the electroacupuncture group was more as compared with the sibutramine group (P<0.01).

CONCLUSIONElectroacupuncture can improve the status of high blood lipids, increase lipoprotein lipase activity, and regulate serum leptin and insulin levels in the obestic rat.

Acupuncture Points ; Animals ; Electroacupuncture ; Humans ; Lipids ; blood ; Obesity ; metabolism ; Rats ; Sodium Glutamate

Objective To study the correlation of patellar stability with age ,knee Kellgren-Lawrence grading and patellar Wiberg typing of adults with Kaschin-Beck disease (KBD) .Methods Through cross-sectional study , we asked 143 adult patients with KBD from several villages in Gansu Province to receive digital radiographic x-ray (DR) which included lateral slice of knee joint and axis slice of patellar .Meanwhile ,each patient had to undergo patellar stability measurement on the DR film which included some indexes such as Insall index , lateral patellofemoral angle ,sulcus angle ,congruence angle ,lateral migrating ratio of patella ,and patellofemoral index . Then ,all the participants were divided into four groups based on the patella Wiberg typing and three groups based on KBD clinical grading .Last ,we analyzed the correlation of patellar stability with patellar Wiberg typing ,clinical KBD grading ,and Kellgren-Lawrence grading of the patients by Spearman test .Results ① By ANOVA test on the patella Wiberg typing ,lateral patellofemoral angle decreased with increased level of patella Wiberg typing ,but congruence angle ,lateral migrating ratio of patella ,and patellofemoral index were increased with increased level of patella Wiberg typing . P values of the six indexes were all <0 .05 ,which indicated that these differences between groups were significant .By ANOVA test on the KBD clinical grading ,lateral patellofemoral angle decreased with increased level of KBD clinical grading ,but congruence angle ,lateral migrating ratio of patella ,sulcus angle ,and patellofemoral index increased with increased level of KBD clinical grading .Besides that P value of Insall index was>0 .05 , P value of the other five indexes were all <0 .05 ,which indicated these differences between groups were significant .② Spearman test :a strong correlation lay between Wiberg patellar typing and other indexes including age ,lateral patellofemoral angle ,congruence angle ,lateral migrating ratio of patella ,and patellofemoral index ( R=0 .684 ,0 .752 ,0 .582 ,0 .533 ,0 .735 , P< 0 .05) .However ,there was no correlation between Wiberg patellar typing and K-L photographical grading (R=0 .030 ,P>0 .05) .K-L grading was strongly correlated with age (R=0 .782 , P< 0 .05 ) , weakly correlated with lateral migrating ratio of patella ( R = 0 .104 , P> 0 .05 ) , but not correlated with the others ( R < 0 .1 , P> 0 .05 ) . Age was strongly correlated with K-L grading and lateral patellofemoral angle (R=0 .782 ,0 .506 ,P<0 .05) ,moderately correlated with congruence angle ,lateral migrating ratio of patella ,and patellofemoral index (R=0 .403 ,0 .361 ,0 .475 , P< 0 .05) ,but not correlated with Wiberg typing ,sulcus angle ,or Insall index ( R=0 .024 ,0 .074 ,0 .053 , P>0 .05) .Conclusion Patellar Wiberg typing is the key factor which influences latellar stability ;age has a limited influence ,while K-L grading does not affect patellar stability .

OBJECTIVETo detect the genes of Neisseria spp. isolated from patients with male genitourinary tract infections, and to study the pathogenicity of non-gonococcal strains of Neisseria and the laboratory diagnosis for the infections caused by Neisseria spp.

METHODSUsing polymerase chain reaction and nucleotide sequencing, we amplified and sequenced 4 genes of Neisseria spp. isolated from patients with male genitourinary tract infections, including 16S rRNA, orfl, cppB and nspA.

RESULTSFourteen Neisseria strains were identified through analysis of the 16S rRNA gene, including 3 N. mucosa strains, 3 N. cinerea strains, 2 N. gonorrhoea strains, 2 N. sicca strains, 2 N. subflava strains, 1 N. lactamica strain, and 1 N. polysaccharea strain. Among them, 9 showed positive results in gonococcal fluorescence-labeled multiplex-PCR detection, 1 in cppB gene reaction, 5 in orfl gene reaction, and 3 in nspA gene reaction. The consistency rate was 85.7% between the above results from our gene detection and those from the routine bacteriological methods.

CONCLUSIONThe cppB gene is absent in the non-gonococcal strains of Neisseria spp. that can cause male genitourinary tract infection. Most of the strains not only lack virulence-associated orfl and nspA genes, but also show positive results in gonococcal fluorescence-labeled multiplex-PCR detection, which is one of the important reasons for the misdiagnosis and missed diagnosis of gonorrhea infection. The combination of routine bacteriological methods and gene detection in laboratory examinations may help improve the accuracy rates of Neisseria species identification and clinical diagnosis of the infections caused by Neisseria spp.

Genes, Bacterial ; Gonorrhea ; diagnosis ; microbiology ; Humans ; Male ; Neisseria gonorrhoeae ; classification ; genetics ; Polymerase Chain Reaction ; Urinary Tract Infections ; microbiology