Objective To assess the influence of PBL formation study effectiveness of TCM students.Methods The controlled studies with the teaching format of both PBL and LBL were included to assess the effectiveness of learning,by searching CBM,CNKI and VIP database.All data was analyzed on Revman 5.1.Results 9 articles were included ; All were low in the quality of their methodology,and the theoretical scores of students in PBL or LBL format shows no statistical difference [SMD=3.76,95％ CI ( -0.62,8.15 ) ].PBL format was superior to LBL on students' practical scores [SMD=7.62,95％ CI ( 3.92,11.32 ) ]; Compared to LBL format,PBL proved to have a better influence on students' self-assessment for learning ability[OR=3.69,95％ CI( 1.88,7.21 )].Conclusion PBL helps to enhance the activeness of students,to improve their practical ability,which is valuable if applied in clinical concerned courses.But the included studies were low in quality; more rigorously randomized controlled teaching trials are expected to verify the conclusion.

A model of classification techniques was established according to the sex and age by quantitative analy-sis of the risk factors for air popplution-induced respiratory diseases using the C4 .5 decision-making tree classifi-cation.The classification-based quantitative analysis was studied.The role of risk factors for respiratory diseases in 9 groups of people was assayed .This method can be used to analyze the risk factors for both respiratory diseases and other diseases in different populations, and can thus provide the evidence for their prevention and control.

Objective To investigate the effects of rhubarb on the expression of glucocorticoids receptor (GR)and peripheral blood lymphocytes in burning-induced septic rats. Methods Sixty-six male healthy Sprague-Dawley(SD)rats were randomly divided into sham operated control group(n=18),sepsis model group(n=24) and rhubarb treatment group(n=24),each group was further randomly divided into 12,24 and 72 hours subgroups according to different time points. The model of scald sepsis was replicated by scald injury induced by boiling water at the rat back accounting for 30% total body surface area(Ⅲ grade of scald),and administration of endotoxin (5 mg/kg)into the peritoneal cavity 12 hours after scald injury. After the successful establishment of septic models, the rats in the rhubarb treatment group were immediately infused with 50 mg/kg rhubarb powder dissolved in 1 mL saline through a gastric tube,while the rats in sham operated control group and sepsis model group received saline by the same way as a substitute for rhubarb. The the binding capacity of GR of peripheral blood leucocyte and binding activity of GR of hepatocyte were analyzed by radiation ligands binding assay. The CD4+,CD8+as well as CD4+/CD8+ ratio in peripheral blood lymphocytes were detected by flow cytometer. Results The binding capacity of GR of peripheral blood leucocyte and binding activity of GR of hepatocyte were significantly decreased in a time-dependent manner in sepsis model group compared to those of the sham operated control group,while in the rhubarb treatment group they were increased in a time-dependent manner after interference of rhubarb, and they were higher than those in the model group at the same time points〔leukocyte GR binding capacity (locus/cell)at 12,24,72 hours :1 515.38±300.44,1 859.63±258.26,1 890.50±307.88 vs. 1 122.63±225.39, 1 008.88±150.41,724.38±91.19;hepatocyte GR binding capacity(fmol/mg):210.19±26.26,258.01±20.98, 283.38±38.21 vs. 153.11±30.07, 129.83±26.89, 94.08±14.30, all P<0.01〕. Compared with the sham operated control group,the CD4+ and CD8+ were decreased in various degrees at 12 hours and 24 hours in the septic group, at 24 hours the differences being statistically significant (P<0.01 and P<0.05). CD4+/CD8+ratios were decreased significantly at all time points,the differences were statistically significant at 24 hours and 72 hours(both P<0.01). The CD4+ T cell and CD4+/CD8+ ratio at all the time points were increased at various degrees in the rhubarb treatment group,and the differences from those in the sepsis model group at 24 hours and 72 hours were statistically significant (1.58±0.69, 1.56±0.49 vs. 1.02±0.41, 1.01±1.68, both P<0.01). Conclusion Rhubarb can modulate the binding capacity of GR of peripheral blood leucocyte and the binding activity of GR of hepatocyte,and via its influence on the number of peripheral leucocytes,the immune dysfunction in the sepsis processes is improved.

ObjectiveTo study HIV-1 DNA levels in different parts of HIV patients during the early stage of antiretroviral therapy.MethodsThe peripheral blood,gut associated lymphoid tissues and lymph nodes samples were collected before and 12 weeks after treatment in regular follow-up HIV-1/AIDS patients in Beijing Youan Hospital ( n =11 ).The average age was 39 years old ( 25 to 55 ).Mononuclear Cells were isolated by density gradient centrifugation and then used DNA extraction kit to extract DNA.Realtime quantitative polymerase chain reaction was used to examine HIV-1 DNA copy-number.Non-parametric test was used to analyse the differences of HIV-1 DNA copy numbers among groups.Results Before treatment,HIV-1 DNA copy-number in both gut associated lymphoid tissues ( 10 714 ± 2043 ) copies/106 cells and lymph nodes (9145 ± 1202) copies/106 cells were higher than that in the peripheral blood (66 ± 8) copies/106 cells ( U =0.00,P ＜0.05 ),There was no significant difference between lymph nodes and gut associated lymphoid tissues (U =46.00,P ＞0.05).After 12 weeks of treatment,HIV-1 DNA copy-number in both gut associated lymphoid tissues (1701 ± 790) copies/106 cells and lymph node (11 591 ± 1781 ) copies/106 cells were higher than the peripheral blood ( 18 ± 3 ) copies/106 cells ( Z =- 2.934,P ＜ 0.05 ).There was a significant reduction of DNA copy-number in gut associated lymphoid tissues and peripheral blood after treatment (Z =- 2.934,P ＜ 0.05 ).Conclusion Gut associated lymphoid tissues and lymph nodes may be important latent reservoirs for HIV-1 DNA.

ObjectiveTo study the effects of small-dose glucocorticoid (GC) on glucocorticoid receptor (GR) and cellular immune function in critical patients.MethodsForty ICU critical patients admitted in Shanghai Changzheng Hospital from March 2007 to March 2009 were enrolled in the study and were divided into GC group and non-GC group according to the use or absence of GC.Blood samples were collected at days 1,7 and 10 after GC treatment to detect GR binding affinity of mononuclear leukocytes (MNLs) and polymorphonuclear leukocytes (PMLs) in the peripheral blood and the CD4/CD8 ratio in the T lymphocytes.The method of GC use was that the hydrocortisone was given intravenously at a dose of 100 mg every eight hours.ResultsGR binding capacity of MNLs at day 1 and 7 showed no statistical difference between the GC and non-GC groups.GR binding capacity of MNLs in the GC group was lower at day 1 and was much lower at day 7 (P ＜ 0.05 ).However,in the non-GC group,it was lower at day 1,but showed significant improvement at day 7 ( P ＜ 0.05 ).The change of GR binding capacity of PMLs was similar to that of MNLs.There was no significant difference of CD4/CD8 ratio between the GC and non-GC group at day 1.The ratio of CD4/CD8 in the non-GC group was significantly higher than that in the GC group at day 10 (P ＜0.05).CD4/CD8 ratio in the GC group showed a slight reduction at day 10,with no significant difference from that at day 1.While,the non-GC group showed a significant increase of CD4/CD8 ratio at day 10 as compared with that at day 1 (P ＜ 0.05 ).ConclusionLow-dose GC plays some role in the negative feedback regulation of GR binding capacity of peripheral blood leukocytes and in the inhibition of cellular immune function.

Objective To understand the carriage of NDM-1 and other carbapenemases in carbapenem-resistant Acinetobacterbaumannii(CRAB)in Jiangxi area,and provide laboratory basis for the prevention and control of healthcare-associated infection (HAI). Methods Sixty-four strains of CRAB isolated from clinical specimens from 3 tertiary first-class hospitals in Jiangxi area from January 2015 to June 2016 were collected,susceptibility to com-monly used antimicrobial agents were detected with Kirby-Bauer method. Carbapenemases and metalloenzyme in CRAB were screened with modified Hodge test and EDTA-disk synergy test respectively,carbapenems gene was de-tected by polymerase chain reaction (PCR),NDM-1-producing Acinetobacterbaumannii (A. baumannii)were per-formed conjugation test.Results The resistance rates of CRAB to ampicillin/sulbactam,ciprofloxacin,gentamicin, and levofloxacin were up to 95.31% ,98.44% ,90.63% ,and 54.69% respectively. The positive rates of modified Hodge test and EDTA-disk synergy test were 76.56% and 96.88% respectively. PCR amplification result showed that 87.50% (n= 56)of CRAB carried OXA-23 and VIM-1 genes,18.75% (n= 12)carried SIM,3.13% (n= 2)car-ried OXA-24,and 26.56% (n= 17)carried NDM-1 . CRAB carrying NDM-1 gene were all from The First Affilia-ted Hospital of Nanchang University,64.70% (11/17)of which were pandrug-resistant strains. Conjugation test re-sult showed that NDM-1-producing strains could transfer NDM-1 gene to recipient strain Escherichiacoli J53,then acquired resistance to imipenem. Conclusion Antimicrobial resistance rates of clinically isolated CRAB in this area are high,OXA-23 and VIM-1 genes are the main carbapenemase genes,NDM-1 gene positive CRAB is detected, and there may be a clonal spread of NDM-1 gene in hospital,effective measures should be taken as soon as possible to prevent and control the spread of NDM-1 positive CRAB.

BACKGROUND:Studies have shown that lung cancer stem cel s can be isolated from lung cancer cel lines. But there are few reports about in vitro isolation, culture and identification of lung cancer stem cel s in patients with lung squamous carcinoma.
OBJECTIVE:To explore the feasible methods of harvesting lung cancer stem cel s from fresh lung cancer tissue in patients with lung squamous carcinoma.
METHODS:Side population cel s were isolated by col agenase digestion, Ficol density gradient centrifugation and Hoechst 33342 solution. The isolated cel s were suspended in conditioned medium for isolated culture. Flow cytometry method was used to detect lung cancer stem cel s based on the cel surface markers CD133 and CD44, and the positive rates of CD133+, CD44+and CD133+/CD44+cel s were recorded.
RESULTS AND CONCLUSION:Cel s adhered at 0.5 hour after incubation;typical cel colony was formed at 4 days of culture;cel s showed paving stone-shape at 7 days in a total number of 10 8. The positive rates of CD133+, CD44+and CD133+/CD44+cel s at passage 4 were increased significantly. These findings indicate that stem cel-like lung cancer cel s were obtained from fresh lung cancer tissue in patients with lung squamous carcinoma, which were stably and rapidly amplified in vitro, laying the foundation for the further study on the heterogeneity and resistance of lung cancer stem cel s in the future.

BACKGROUND:Studies have shown that lung cancer stem cels can be isolated from the lung cancer cel lines, But there are few reports on in vitro isolation, culture and identification of lung cancer stem cels in patients with lung squamous carcinoma.
OBJECTIVE:To establish the feasible methods of harvesting lung cancer stem cels from fresh lung cancer tissues in patients with lung squamous carcinoma, and to investigate the alterations in cel number and function during primary culture.
METHODS: Side population cels were isolated by colagenase digestion, Ficol density gradient centrifugation and Hoechst 33342 efflux properties. The isolated cels were isolated and cultured in conditioned medium. Flow cytometry method was used to detect lung cancer stem cels based on the cel surface markers CD133 and CD44, and the positive rates of CD133+, CD44+ and CD133+/CD44+ were recorded. The single cel clones assay, flat colony formation assay and the cel sphere formation assay were used to identify the stem-like characteristics of lung cancer stem cels between the first and fourth generations.
RESULTS AND CONCLUSION:The positive rates of CD133+, CD44+ and CD133+/CD44+ cels at the fourth generation were increased significantly, and the positive rates of CD133+ and CD133+/CD44+ cels at passage 4 were significantly higher than those at the first generation. The abilities of single cel clone formation, the flat colony formation and the cel sphere formation in the fourth-generation cels were greatly enhanced compared with the first-generation cels. Experimental findings showed that stem cel-like lung cancer cels were obtained from fresh lung cancer tissue in patients with lung squamous carcinoma, which stably and rapidly amplified in vitro, laying the foundation for the further study of the heterogeneity and drug resistance of lung cancer stem cels.

AIM:To explore the molecular mechanism through which curcumin reverses hepatocyte growth factor (HGF)-induced resistance to gefitinib in lung cancer cells.METHODS:The methods of MTT assay, wound healing assay and Western blot were used to observe the effects of HGF, curcumin and gefitinib on the migration, drug susceptibility, epithelial-mesenchymal transition, and related signaling pathways in the PC9 lung cancer cells.RESULTS:HGF reduced susceptibility of the PC9 cells to gefitinib, and curcumin significantly reversed HGF-induced resistance to gefitinib.HGF induced migration and epihelial-mesenchymal transition, and promoted c-Met/AKT/mTOR pathway activation in the PC9 cells.Gefitinib alone did not prevent the above activities.However, combined with curcumin, gefitinib prevented the above activities.CONCLUSION:Curcumin reverses HGF-induced resistance of the PC9 cells to gefitinib by preventing epithelial-mesenchymal transition and inhibiting c-Met/AKT/mTOR activation.

Objective To investigate the expression of CD40,CD40L and MMP9 in carotid atherosclerotic plaques and evaluate their roles in carotid atherosclerotic plaque stability.Methods Carotid atherosclerotic plaques were isolated in carotid eversion endarterectomy (GEE) in 37 patients with high-grade stenosis (>70%) including 20 stroke (A group) and 17 non-stroke patients (B group).The control group included samples of normal carotid artery from 11 normal individuals,The RNA expression levels of CD40,CD40 L and MMP9 in all A,B and control groups were quantitatively detected by real-time quantification polymerase chain reaction (PCR) and the protein expression levels were detected by Western blotting analysis.The expression and distribution of CD40,CD40L and MMP9 in carotid atherosclerotic plaques were detected by immunohistochemistry staining.Then correlations between CD40-CD40L and MMP9 were statistically analyzed.Results The relative CD40 mRNA level in high-grade stenosis of A group,B group and normal control were 2.41±0.43,1.03±0.38 and 0.31±0.12,respectively,and MMP9 mRNA 6.88±1.57,1.90±0.44 and 0.39±0.12,respectively.The levels of CD40 and MMP9 mRNA in A group were significantly higher than those in B group (P=0.000),the levels of CD40 and MMP9 in B group were significantly higher than those in controls (P=0.000).There was a linear correlation between CD40 and MMP9 mRNA (r=0.929,P=0.000).However,there were no significantly difference in mRNA levels of CD40L between carotid atherosclerosis and controls.The protein expression levels of CD40,CD40L and MMP9 in A group were significantly higher than those in B group (FCD40=104.100,P=0.000;FCD40L=129.932,P=0.000;FMMP9=13.565,P=0.021) and B group higher than normal controls (FCD40=115.848,P = 0.000;FCD40L= 30.482,P=0.005;FMMP9=35.557,P=0.004).The areas of positive staining of CD40,CD40L and MMP9 in immunochemistry study in A group were significantly higher than those in B group and B group was significantly higher than controls.There were linear correlations between positive staining areas Of CD40 and CD40L,CD40 and MMP9,CD40L and MMP9 (r=0.963,0.959,0.929,P=0.000).Expressions of CD40,CD40L and MMP9 were significantly higher in the shoulder areas of the atherosclerotic plaques than in other areas.Conclusions The CD40-CD40L has an important role in the formation of carotid atherosclerosis and plaque instability,probably by up-regulating MMP9.The expression of CD40L may be regulated by post-transcriptional modification to exert biological effects.