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.

Objective To evaluate the clinical significance of quantitative sensory testing (QST) in screening diabetic peripheral neuropathy of the early stage. Methods One hundred patients with type 2 diabetes mellitus were examined by nerve conduction velocity (NCV) and QST examination. With the NCV positive as the gold criterion for screening diabetic peripheral neuropathy of the early stage, the sensitivity and specificity of QST was further analyzed for diagnosis of the early stage diabetic peripheral neuropathy. Results Among the 100 patients with type 2 diabetes mellitus,there were 41 cases positive and 59 cases negative in NCV examination. On the other hand,there were 74 cases positive,and 26 cases negative in QST. The sensitivity and specificity of QST for the diagnosis of early stage diabetic peripheral neuropathy was 97.56% (40/41) and 42.37% (25/59). Conclusions In the screening of early stage diabetic peripheral neuropathy,QST shows higher detection sensitivity,but lower specificity than NCV examination. Therefore, QST may be an examination for the supplement of the routine electromyography.

Objective To explore virtual three-dimensional (3D)model for intravascular interventional devices,the method of preoperative simulation and its value in clinical work.MethodsThe virtual models including catheter,guide wire,stent and coil were established by using the 3D moulding software of 3D Studio MAX R3. The interventional preoperative simulation was performed on personal computer including 21 patients of cerebral aneurysm embolization ( anterior communicating artery 5,posterior communicating artery 10,middle cerebral artery 3,internal carotid artery 2,and vertebral artery 1 ),during interventional procedures,the surgeon relied on the simulation results for plastic micro-guide wire,catheter and the release of micro-coils and stents.Results( 1 ) All the virtual instruments and real instruments had similar shape,the overall tine for constructing virtual model was about 20 hours.The preoperative simulation took 50 to 80 minutes. (2) The simulation result of catheter insertion in the 18 cases had relevant value to guide micro-catheter,molding micro-guide wire tip,and shortened the operating time.For embolization,the simulation results of filling coil and releasing stent were similar to surgical results in 76％ of the patients (16/21).(3)For teaching and training,93％ (38/41) of doctors in training believed that preoperative simulation facilitated the understanding of surgery. ConclusionsThe method of virtual model of intravascular interventional devices was reliable.The preoperative simulation results could be used to guide practical clinical operation with relatively high degree of similarity,and could play a role in promoting researches on interventional virtual operations.

Objective To evaluate ultrasound-guided methylene blue dyeing for radical liver segmental resection.Methods Liver segmental resection with uhrasound-guide methylene blue dyeing (UMD-SR) was performed in 16 cases, results were compared with 16 conventional liver segmental resection (CSR) retrospectively.Results All the operations under uhrasound-guided methylene blue dyeing were successfully carried out, among them, reverse dyeing was used in cases with segment Ⅳ, Ⅴ, Ⅷ resection.The blood loss in UMD-SR group was much less than CSR group(t =3.011 ,P =0.009) , at the cost of a longer operation time (t =5.423,P =0.000 07).There was no difference in the mortality and morbidity rates between two groups.Tumor recurrence rate was 6.25％ in UMD-SR group and 18.75％ in CSR group (x2 =0.133,P =0.285).Conclusions Ultrasound-guided methylene blue dyeing liver segmental resection can reduce the blood loss during operation, improve the safety of hepatectomy in case of hepatic carcinoma.

Previous studies showed that three methods for regulating and invigorating lung and kidney (lung invigorating and spleen strengthening, lung invigorating and kidney tonifying, and Qi supplementing and kidney nourishing) could regulate inflammatory signaling pathways of chronic obstructive pulmonary disease (COPD) in rats, so as to alleviate inflammation. In the present study, R-value comprehensive evaluation method was used to evaluate the comprehensive effect of three methods for regulating and invigorating lung and kidney on inflammatory signaling pathways. Rats were randomly divided into control, model, lung invigorating and spleen strengthening, lung invigorating and kidney tonifying, Qi supplementing and kidney nourishing and aminophylline groups. The COPD rat models were established by cigarette smoking combined with bacterial infection, and orally administered with drugs between the 9th and 20th week. Afterwards, efforts were made to observe the long-term effects between the drug withdrawal and the 32rd week and detect indicators in two batches in the 20th week and 32th week. Specifically, (1) Linking JAK/STAT signaling pathway: JAK2 mRNA, and protein expressions of STAT-1, STAT-3, STAT-5, JAK-2; (2) NF-kappaB signaling pathway: Smad2 mRNA and protein expressions of I-kappaB, NF-kappaB, TGF-beta1; (3) PPARgamma and antioxidant signaling pathway: SOD, PGE mRNA, PPARgamma protein. According to the results, 5 indicators in JAK/STAT pathway, 4 indicators in NF-kappaB pathway, and 3 indicators in PPARgamma pathway were significantly rectified by three methods for regulating and invigorating lung and kidney in between the 20th week and 32nd week. Between the 20th and 32nd week, the recipes for rectifying JAK/STAT pathway with intensity from high to low were recipes for lung invigorating and spleen strengthening, Qi supplementing and kidney nourishing, lung invigorating and kidney tonifying, aminophylline, particularly those for lung invigorating and spleen strengthening; The recipes for rectifying NF-kappaB pathway with intensity from high to low were recipes for lung invigorating and spleen strengthening, lung invigorating and kidney tonifying, Qi supplementing and kidney nourishing and aminophylline, particularly the first three types of drugs. The recipes for rectifying PPARgamma and antioxidant signaling pathway with intensity from high to low were recipes for lung invigorating and kidney tonifying, Qi supplementing and kidney nourishing, lung invigorating and spleen strengthening and aminophylline. Therefore, three methods for regulating and invigorating lung and kidney showed better long-term effects in regulating COPD lung inflammation signaling pathways. Specifically, recipe for lung invigorating and spleen strengthening showed a better effect in JAK/STAT and NF-kappaB pathways, while recipe for lung invigorating and kidney tonifying and Qi supplementing and kidney nourishing showed better effects in PPARgamma and antioxidant signaling pathways. In conclusion, R-value comprehensive evaluation method can evaluate the comprehensive effect of medicines and define the ranking of multiple drugs and their main targets.
Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Kidney ; drug effects ; physiopathology ; Lung ; drug effects ; immunology ; metabolism ; physiopathology ; NF-kappa B ; immunology ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; immunology ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Smad2 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo compare postoperative blood loss under different negative pressures of drainage after total hip arthroplasty for the treatment of femoral neck fractures.

METHODSFrom January 1st to December 30th 2013, 74 patients with femoral neck fractures treated with total hip arthroplasty were randomly divided into two groups: high negative pressure drainage group and low negative pressure drainage group. In high negative pressure drainage group, there were 34 cases including 10 males and 24 females, with a mean age of (75.94 ± 9.02) years old, and the patients were treated with 60 kPa negative pressure of drainage. In the low negative pressure drainage group, there were 40 cases including 13 males and 27 females, with an average age of (74.93 ± 8.90) years old, and the patients were treated with 30 kPa negative pressure of drainage. The amount of total drainage, total blood loss, and hemoglobin change were compared between these two groups.

RESULTSAll the patients got primary healing without infections. In high negative pressure drainage group,the change of hemoglobin was (41.74 ± 15.69) g/L, total blood loss was (1,217.73 ± 459.50) ml and the drainage volume was (312.94 ± 103.44) ml; while in low negative pressure drainage group,the results were (34.90 ± 12.90) g/L, (904.01 ± 381.58) ml and (129.25 ± 44.25) ml separately. All the results in high negative pressure drainage group were higher than those in the other group. Three days after operation, the change of hemoglobin was (46.00 ± 13.29) g/L and total blood loss was (1,304.72 ± 421.75) ml; while in low negative pressure drainage group, the changes of hemoglobin was (43.87 ± 11.39) g/L and total blood loss was (1,196.78 ± 344.20) ml; there were no statistically significant differences between two groups.

CONCLUSIONWhen placing drainage devices after total hip arthroplasty for the treatment of femoral neck fractures, the level of negative pressure should be chosen according to preoperative level of hemoglobin and HCT in patients. For old patients with femoral neck fracture, low negative pressure is more suitable.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; methods ; Case-Control Studies ; Female ; Femoral Neck Fractures ; surgery ; Humans ; Male ; Middle Aged ; Negative-Pressure Wound Therapy ; Postoperative Hemorrhage ; prevention & control