Objetive To investigate a method of collecting lung cancer cells with bronchofibroscopic biopsy for primary culture and to improve the success rate of primary culture. Methods Thirty lung cancer specimens were obtained through bronchoscopic biopsy for primary culture. The correlation of cancer morphology under bronchofi-broscopy and success rate of primary culture was analyzed. Results Among the lung cancer specimens obtained through bronchoscopic biopsy, primary culture was successful in 17 of 30 cases (56.67%) . The success rate of cauliflower-like tumor mass under bronchofibroscopy was 84.62% (11/13) . The success rate of infiltrating tumor mass under bronchial mucosa with luminal stenosis with or without cristate were 66.67% (2/3) and 37.5%(3/8), respectively. The primary culture of a globular and stiff tumor mass was successful only 1 in 6 cases (16.67%) .Conclusions The primary culture of lung cancer cells obtained from bronchofibroscopic biopsy is simple and effective with a total success rate of 56.67%. Furthermore, the success rate of primary culture is signifi-cantly correlated with the cancer morphology under bronchofibroscopy.

This paper summarize the practice in the teaching reform of microbiology experiment in recent years. We identify the main contents of experimental teaching systems and pay much more attention to peo-ple-oriented. Through the reform of teaching and assessment methods,students are trained to cultivate their practical ability and spirit of innovation.

AIM: To investigate whether RNA interference (RNAi) induced by small interference RNA (siRNA) could suppress Polo- like kinase- 1 (Plk 1 ) expression and its effects in A549 cells. METHODS: A recombinant plasmid containing siRNA targeting Plk1 ( psiRNA - hH1 - Plk1 ) was transfected into A549 cells with Lipofectamine 2000.Expressions of Plk1, cyclin B1 and p53 protein were detected by Western blotting. Cell proliferation was evaluated by direct cell counting, while cell cycle and apoptosis were examined by flow cytometry, and expression of α - tubulin was detected by immunofluorescence. RESULTS: The results demonstrated that sequence specific siRNA targeting Plk1 was capable of suppressing Plk1 expression, and reflecting in lower kinase activity in A549 cells. The level of Plk1 protein was reduced by at least 70％ after 48 h of psiRNA - hH1 - Plk1 treatment relative to controls. Expressions of cyclin B1 and p53 were increased greatly after Plk1 depletion, and cells showed absence of microtubule polymerization and spindle abnormalities in staining for α -tubulin. Growth inhibition, G2/M arrest and apoptosis were observed in psiRNA -hH1 -Plk1 transfected group. CONCLUSION: All these data suggest that siRNA targeted against human Plk1 may be a valuable tool in cancer therapy.

Objective To explore the changes and clinical significance of levels of serum vascular endothelial growth factor(VEGF)and endostatin in smokers. Methods In a case-control study,levels of serum VEGF and endostatin were determined in 82 smokers with lung cancer,82 pair-matched smokers without lung cancer and 20 healthy non-smokers by enzyme-linked immunoabsent assay(ELISA) or competitive enzyme immunoassay.Results The level of serum VEGF in smokers with lung anncer[(16.1±7.9)ng/ml]was markedly higher than that in the other two groups(both P＜0.01).The level of serum VEGF in smokers without lung cancer was significantly higher than that in healthy non-smokers(P＜0.05).The level of serum endostatin in smokers with lung cancer was significantly higher than that in healthy nonsmokers(P＜0.01),but was not significantly different from that in smokers without lung cancer(P＞0.05),and that in smokers without lung cancer was significantly higher than that in healthy non-smokers (P＜0.05).Notably,the ratio of endostatin to VEGF in smokers with lung cancer(1.3±0.5)was significantly lower than that in the other two groups(both P＜0.01).However.there was no significant difference in it between smokers without lung cancer and healthy non-smokers(P＞0.05).The level of serum VEGF correlated significantly to that of endostatin in smokers both with and without lung cancer(P＜0.01).Conclusions These findings suggest that smoking may result in imbalance of levels of serum endostatin and VEGF leading to tumorigenesis.The ratio of endostatin to VEGF can be used as an early diagnostic indicator for lung cancer in smokers.Periodic determination of levels of serum VEGF and endostatin as well as the ratio of endostatin to VEGF is of clinical importance.

OBJECTIVETo analyze the parameters of urodynamic tests for patients with type-III B prostatitis and evaluate the significance of the results of urodynamic tests in the choice of therapies for this disease.

METHODSUrodynamic tests were performed for 87 type-III B prostatitis patients aged 22-45 (30.7 ± 8.5) years, who had moderate or severe lower urinary tract symptoms (LUTS) and failed to respond to routine therapy. Different treatments were administered according to the results of urodynamic tests followed by observation of the therapeutic effects.

RESULTSUrodynamic abnormalities were found in 70 of the 87 patients, bladder outlet obstruction in 28 (32.2%), detrusor overactivity in 25 (28.7%), bladder hyperesthesia in 18 (20.7%), low compliance in 10 (11.5%), detrusor-external urethral sphincter dyssynergia in 1 (1.1%), and impaired detrusor contractile function in 1 (1.1%). Treatments achieved obvious effectiveness in 26 cases (29.9%), effectiveness in 51 (58.6%), and no effectiveness in 10 (11.5%).

CONCLUSIONUrodynamic tests contribute significantly to the choice of therapies for type-III B prostatitis patients with moderate or severe LUTS.

Adult ; Humans ; Lower Urinary Tract Symptoms ; physiopathology ; therapy ; Male ; Middle Aged ; Prostatitis ; physiopathology ; therapy ; Urethra ; physiopathology ; Urinary Bladder Neck Obstruction ; physiopathology ; Urinary Bladder, Overactive ; physiopathology ; Urodynamics

OBJECTIVETo observe the primary prevention role of Wuling Capsule (WC) on poststroke depression (PSD) patients.

METHODSAcute stroke patients were recruited and randomized into 2 groups by stratification, 55 in each group. All patients received same routine treatment of cardiovascular diseases. Patients in the experimental group additionally took WC (0.33 g each pill), 3 pills per day, three times per day; while those in the control group additionally took placebos, 3 pills per day, three times per day. Two weeks consisted of one therapeutic course. The diagnosis of PSD was performed once every other week. Those in accordance with PSD diagnosis discontinued any drug therapy. Those not in accordance with PSD diagnosis continued the drug therapy for 1-12 therapeutic course(s) (in total of 6 months). If they were still not in accordance with PSD diagnosis, then they discontinued the drug therapy. The morbidity of PSD, the average time of depression occurrence, Hamilton depression rating scale (HAMD) score, and adverse reactions were observed.

RESULTSThe 1-, 3-, and 6-month morbidity of PSD was 8%, 16%, and 34% in the experimental group, while they were 19.6%, 29.4%, and 54.9% in the control group. The occurrence rate was lower in the experimental group than in the control group. Besides, there was statistical difference in the 6-month occurrence rate between the two groups (chi2 = 4.465, P < 0.05). The average time of PSD occurrence was longer in the experimental group than in the control group (14.96 +/- 8.31 weeks vs. 9.36 +/- 6.06 weeks; t=6.762, P < 0.05). The HAMD score at the PSD occurrence was 11.96 +/- 2.14 in the experimental group, lower than that of the control group (14.57 +/- 4.24), showing statistical difference (t=5.641, P < 0.05).

CONCLUSIONWC was superior to the placebos in lowering the incidence of PSD, delaying the occurrence time of PSD, attenuating the depression degree of PSD, and had certain preventive effect on the incidence of PSD.

Aged ; Capsules ; Depression ; etiology ; prevention & control ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Primary Prevention ; Stroke ; complications

The changes of bioactive constituents were analyzed for Puhuang-Wulingzhi before and after compatibility and the antiplatelet and antithrombin activitives were evaluated in order to elucidate the scientific and reasonable of Puhuang-Wulingzhi compatibility. UPLC-QTOF-MA-Markerlynx, principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis were used for data analysis and tracking changes of chemical composition during the decocting process. In vitro platelet aggregation induced by ADP, thrombin time(TT) and prothrombin time (PT) were investigated for Puhuang-Wulingzhi before and after compatibility. The results showed that significant differences were found between the mixed decoction and codecoction of Wulingzhi and Puhuang. Five compounds changed obviously were identified as typhaneoside, naringenin, isorhamnetin-3-O-ruinoside, quercetin-3-O-neohesperidoside, kaempferol-3-O-neohesperidoside. The codecoction, comparing with the single decoction, was more significant in antiplatelet aggregation and could prolong thrombin time. In the same crude drug dose, the thrombin time (TT) elongation were greater. These data could provide references for elucidation of bioactive components for this herb pair.
Animals ; Antithrombins ; chemistry ; pharmacology ; Blood Platelets ; drug effects ; physiology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Humans ; Molecular Structure ; Platelet Aggregation ; drug effects ; Rabbits ; Thrombin Time

Objective To investigate ATM deletion [del (ATM)] in chronic lymphocytic leukemia (CLL) and study its correlation with the clinical stage. Methods Spectrum Orange~(TM) labeled sequence specific DNA probe for ATM locus on 11q22.3 and fluorescence in Situ hybridization (FISH) was used to examine del (ATM) in 28 newly diagnose patients with CLL. FISH analysis were performed on bone marrow smears. Clinical staging was done according to Binet Method.Fisher exact propability was used to study the relations between del (ATM) and clinical feature. Results 4 out of the 28 cases were found with deletion of ATM. The incidence of del (ATM) in BinetA, BinetB and BinetC was 1/9 (11.1 %), 1/8 (12.5 %), 2/11 (18.2 %), respectively. Fisher exact propability showed that deletion of ATM was not associated with its clinical feature. Conclusion Application of FISH on archival bone marrow smears is a simple, liable method, and can be readly used to retrospective study of clonal blood system diseases. Deletion of ATM was common cytogenetic changes in CLL patients.And the significance of del (ATM) in the prognosis of CLL in China needs to be further investigated.

Objective To evaluate the immunogenicity and immunoprotective effect of heat shock protein 60 kDa (SjHSP60) of Schistosoma japonicum in mice after immunization and challenge infection, and explore the mechanism. Methods B cell/an?tibody?related databases and analysis tools were used to predict B?cell epitopes of SjHSP60. The mice were immunized with the recombinant SjHSP60 and challenged with S. japonicum cercariae. SjHSP60?specific antibodies in serum were detected by ELI?SA. The level of splenocyte proliferation was determined by 3H?TdR incorporation. Ex vivo suppression assay was performed to in?vestigate the effects of CD4 +CD25 + regulatory T cells (Tregs) induced by SjHSP60. Results SjHSP60 possessed multiple pre?dominant regions of B?cell epitopes. SjHSP60 induced a significant increase in both SjHSP60?specific IgG levels (P < 0.01) and splenocyte proliferation (P < 0.01) with a higher IFN?γ production (P < 0.01). However, the immunization with SjHSP60 resulted no significant reduction in adult worms (P > 0.05) and liver?accumulated eggs (P > 0.05) in S. japonicum?infected mice. Ex vivo assay showed that CD4+CD25+ Tregs from SjHSP60?immunized mice enhanced immunosuppressive activity. Conclusion SjH?SP60 has a dual role in host immune system, being involved in the induction of dominant humoral and cellular immune responses as well as in the enhancement of immunosuppression.

Objective To detect expression of TEL-AML1 fusion genes in pediatric cases with acute lymphoblastic leukemia(ALL) and discuss the role of reverse transcriptase polymerase chain reaction(RT-PCR)and fluorescence in situ hybridization(FISH) in detection of t(12 ;21) and the clinical significance. Methods TEL-AML1 fusion gene was identified in bone marrow munonuclear cells from 31 newly diagnosed childhood ALL patients by NRT-PCR, FISH and conventional cytogenetic analysis (CCA). Results TEL-AML1 fusion gene was found in 7 out of 31 cases, accounting for 22.6 % in pediatric ALL, and 7 out of 31 cases accounting for 25.9 % in B-ALL Seven cases were found with t (12;21) by FISH and NRT-PCR. The incidence of the t(12;21) was 22.6 % in newly diagnosed pediatric ALLs. Conclusion It is concluded that TEL-AML1 rearrangement is a frequent molecular abnormality in childhood ALL. t(12;21) is the most common cytogenetic translocations in Chinese pediatric ALLs, but it is always difficult to identify by routine CCA.Other molecular methods, e.g. NRT-PCR and FISH are powerful in detecting such a critical genetic translocation.