Objective: To investigate the inhibitory effect of short hairpin RNA (shRNA) -mediated silence of phospholipase C epsilon (PLC)̇gene expression on the proliferation of renal carcinoma 786-0 cells and its action mechanism. Methods: Liposome was employed to mediate the transfection of both the recombinant plasmid (pGenesil-PLC)̇and the control plasmid (pGenesil-NP) into the 786-0 cells. RT-PCR was used to detect the PLĊ mRNA expression after being transfected for 48 h. MIT assay was conducted to detect the proliferation inhibitory rate at 24, 48, and 72 h after transfection, respectively. Flow cytometry (FCM) was performed to analyze cell cycle after 48-h transfection. RT-PCR and immunocytochemistry were used to analyze the expression levels of both p27 and Ki67. Results: The expression of PLĊmRNA was significantly inhibited by recombinant plasmid transfection with the inhibitory rate of 69.7%. The proliferation inhibitory rates were 21.2%, 31.6%, and 32.7% after being transfected for 24, 48, and 72 h, respectively. FCM analysis demonstrated that the distribution of cell cycle changed. The number of cells in the G0/G1 phase increased, and that in both the S phase and G2/M phase decreased. Cells were arrested at the G0/G1 phase, and the subdiploid "apoptotic peak" appeared at the same time. RT-PCR and immucytochemistry indicated that the expression of p27 was up-regulated and that of Ki67 was downregulated. Conclusion: The proliferation of the renal carcinoma 786-0 cell line is inhibited by interference of PLĊgene expression, and the underlying mechanism may partially be related with up-regulation of P27 and down-regulation of Ki67.

Objective To explore the clinical features of pulmonary tuberculosis complicated with DM and related factors. Method The manifestations of 87 newly diagnosed PTB patients with type 2 DM(DM-PTB group) at two hospitals in Tianjin during October 2003 to October 2004 were compared to 88 PTB patients without DM who were randomly choosen from the same hosipital.The factors related to sputum-positive TB or cavity were explored through unconditional logistic regression model. Results DM-PTB patients showed significanly lower frequencies of cough(66.7% vs 83.0%),toxicity symptoms(37.9% vs 62.5%) and haemoptysis(16.1% vs 33.0%) than those of PTB group.Positive-sputum(63.2% vs 34.1%),cavity(56.3% vs 13.6%),wide lesions and bilateral lesions on chest radiographs was more common in DM-PTB group than in PTB one.Introversion(OR=1.582, 95%CI:1.063 ～2.355),type 2 DM(OR =2.186,95% CI:1.082 ~4.420)and cavity(OR =2.793,95% CI:1.308 ～5.964)were factors independently associated with positive-sputum TB.The factors related to cavity included diabetes(OR=14.199,95%CI:5.376~ 37.501),cough,postive sputum and lore-up lobe lesion.Conclusion This study confirmed that clinical manifestations and chest radiographs of PTB patients complicated with type 2 DM significantly departed from the typical presentation.Type 2 DM seems to have a negative effect on postive-sputum and cavity.So it should be alarmed that DM-PTB patients who tend to have untypical clinical symptom and higher rate of positive sputum become the reservoirs of infection to cause the transmission of TB.

AIM: To investigate the role of autophagy in the apoptosis of human pulmonary artery endothelial cells (HPAECs) induced by cigarette smoke extract (CSE).METHODS: HPAECs were cultured routinely.HPAECs were treated with CSE at different concentrations, and the cell viability was detected by MTT assay.HPAECs were divided into control group, CSE group, 3-methyladenine (3-MA) group and 3-MA+CSE group.The autophagy was observed under fluorescence microscope with monodansylcadaverine (MDC) staining.Annexin V/propidium iodide staining and Hoechst 33342 staining were employed to detect apoptosis.In addition, the protein levels of LC3, beclin-1 and cleaved caspase-3 were determined by Western blot.RESULTS: MDC staining showed the increased production of autophagic vacuoles was observed in CSE group.The results of Western blot showed that the expression levels of autophagy-related proteins LC3 and beclin-1 were increased, while 3-MA pretreatment inhibited the expression of these proteins and the production of autophagic vacuoles.Observation with Annexin V/propidium iodide staining and Hoechst 33342 staining showed that the apoptotic rate in CSE group was significantly higher than that in control group, and pretreatment with 3-MA induced further increase in the cell apoptosis.The protein level of cleaved caspase-3 in CSE group was significantly higher than that in control group (P<0.05), and 3-MA+CSE treatment induced the further increase in the protein level of cleaved caspase-3.CONCLUSION: CSE induces autophagy and apoptosis in the HPAECs.Inhibition of autophagy promotes the apoptosis induced by CSE in HPAECs, which can be achieved through activation of caspase-3.

Animals ; Blood Glucose ; drug effects ; Body Weight ; drug effects ; Chronic Disease ; Female ; Hypoxia ; physiopathology ; KATP Channels ; drug effects ; Male ; Propylamines ; pharmacology ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective To investigate the anti-tumor effect of ZM-66 on multidrug-resistant leukemic cell line K562/ADM.
Methods The K562/ADM cells were treated with varying concentrations (0, 1, 2, 4×10-3 mmol/L) of ZM-66 or etoposide for 24 hours. The proliferation was detected by Sulforhodamine B Sodium Salt (SRB) assay and apoptosis was detected by flow cytometry analysis and fluorescent staining. In addition, the expression levels of p53 and bax genes in K562/ADM cells were detected by RT-PCR analysis. The level of P-glycoprotein (P-gp), P53 and Bax protein in K562/ADM cells were detected by Western blot assay.
Results SRB assay demonstrated that etoposide had little inhibitory effect on K562/ADM cells, whereas ZM-66 (1, 2, 4×10-3 mmol/L) had significantly inhibitory effect on K562/ADM cells (all P<0.01). The acridine orange/propidium iodide dual staining showed that there were typical condensation of chromatin and nuclear fragmentation nuclei with red color in ZM-66 treated cells. Flow cytometric analysis showed that there was a significantly increase of apoptotic cells in K562/ADM cells after treated with ZM-66. RT-PCR showed that the p53 and bax mRNA expression levels in K562/ADM cells treated with ZM-66 at 1, 2, 4×10-3 mmol/L were higher than those in the cell without treatment. Western blot showed that the P53 and Bax protein expression levels in K562/ADM cells treated with ZM-66 at 2, 4×10-3 mmol/L were higher than those in the cell without treatment. But the P-gp protein expression level in K562/ADM cells treated with ZM-66 at 2, 4×10-3 mmol/L was gradually lower than those in the cell without treatment.
Conclusion ZM-66 is able to induce cell death by apoptosis in vitro, as a result of the reverse of the apoptosis resistance in drug-resistant K562/ADM cells by modulating expression of key factors associated with apoptosis induction.

A bacterium strain BFHM2002 is isolated from Lake Donghu, Wuhan. BFHM2002 has advantages that it can produce melanin with a high rate and high yield in the absence of tyrosine. Induced by tyrosine, melanin yield can be dramatically increased. BFHM2002 may be identified as a new strain in Bacillus firmus, for melanin-production.

The original strain was treated with UV combining LiCl. The total carotenoid yield of the strain UL-61 was 7.62mg/L, the maximal yield of astaxanthin had reached 616.8?g/g under 25℃. 8 variables including fermentation temperature, initial pH were optimized by Plackett-Burman design and Response Surface Analysis of SAS software. The temperature, initial pH and corn steep liquor were the major factors. The optimal conditions were predictted to be 16.78℃, pH4.73 and 7.06 mg/L, respectively. Under these conditions, the theroretical maximal yield of carotenoid was 3.9407 mg/L and the practical maximal yield was about 3.9261mg/L. The carotenoid production was increased by 20.4% when Phaffia rhodozyma was cultivated under the optimal conditions as compared with the control.

Objective To investigate the relationship between high-sensitive C-reactive protein(hsCRP)and insulin resistance(IR)in elderly hypertension patients.Methods The levels of ambulatory blood pressure mo- nitoring(ABPM),fasting plasma glucose(FPG),fasting insulin(FINS),von Willebrand factor(vWF),hsCRP were measured in elderly patients with hypertension alone(n=260),hypertension coexit with diabetes(n=230), and healthy matched subjects(n=250).Results The levels of FINS,FPG,vWF,hsCRP,systolic blood pres- sure(SBP),diastolic blood pressure(DBP)in the HT and HT+DM group were significantly higher than that in the NC group(P

Melatonin has been reported to inhibit hepatic fibrosis and the mechanism may be correlated to its anti-oxidant effect.Nevertheless,the mechanism is not completely identified.This study was conducted to investigate the effects of melatonin on TGF-β1/Smad signaling pathway in liver fibrosis in rats.The liver fibrosis model was made by the subcutaneous injection of CCl4.The liver pathology changes were detected using hematoxylin and eosin (H&E) staining and Van Gieson (VG) staining.Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were measured with an autoanalyzer.Glutathione peroxidase (GPx) activities and levels of malondialdehyde (MDA) and hydroxyproline (Hyp) in liver were evaluated by spectrophotometry.Expression levels of TGF-β1,Smad2/3,phosphorylated Smad2/3 (p-Smad2/3) and Smad7 in liver were detected by immunohistochemistry and Western blot analysis.Results showed that melatonin suppressed CC14-induced liver fibrosis,along with an improvement in histological changes,significant decreases in pathologic grading sores and obvious decreases in Hyp levels in liver.Melatonin improved liver function indicated by decreased serum ALT and AST activities.In addition,melatonin exerted its anti-oxidant effects,as supported by decreased MDA levels and increased GPx activities in liver.Furthermore,melatonin inhibited TGF-β1/Smad pathway,as evidenced by decreased TGF-β1,Smad2/3 and p-Smad2/3 expression and increased Smad7 expression in liver.In conclusion,melatonin may suppress CCl4-induced hepatic fibrosis in rats via inhibiting TGF-β1/Smad pathway.It is possible for melatonin to be a potential reagent to treat and cure liver fibrosis.

To analyze the clinical characteristics and combined use of chemical and traditional Chinese medicine (TCM) medicine of hospitalized patients with psoriasis base on real world database, 2 991 cases of hospitalized patients with psoriasis in hospital information system (HIS) database from 16 hospitals in China were analyzed for general hospitalization information, combined diseases and combined use of drugs et al. The results showed that half of inpatients aged 18-45 years old. The most common syndrome of TCM was intrinsic blood heat. More than 1/3 inpatients' hospitalization time was 18-25 days, and the average expense of hospitalization was 6 989. 20 RMB. The top five combined diseases were hypertension, non-alcoholic fatty liver disease, diabetes, upper respiratory tract infection and lipoprotein disorders. Medicine information analysis showed 599 chemical medicines and 341 TCMs were used and combined use of drugs was common in clinical practice. Licorice extract medicine was the most common combined TCM with western medicine; in the next two places were compound Qingdai capsule and tripterygium glycosides. The most common combined use of chemical medicines were Vitamin C, calcium gluconate, ketotifen, cetirizine, retinoic acid and external use glucocorticoid. Anti-inflammatory and liver protection, clearing heat and toxic materials, activating blood and dissolving stasis were the most common combined TCM medicine with western medicine, while the most common combined chemical medicine with TCM were anti-allergic, anti-infection, glucocorticoid and retinoic acid. In conclusion, half of hospitalized patients of psoriasis were young adults. The main type of combined diseases was metabolic disorders and upper respiratory infections. Combined use of chemical medicine and TCM was common in clinical practice. Licorice extract medicine was the most common combined TCM with western medicine.
Adolescent ; Adult ; Aged ; Ascorbic Acid ; therapeutic use ; Calcium Gluconate ; therapeutic use ; Cetirizine ; therapeutic use ; China ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Ketotifen ; therapeutic use ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Psoriasis ; drug therapy ; Tretinoin ; therapeutic use ; Young Adult