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

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

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.

Aim To study mRNA and protein expression of eNOS in pulmonary tissue of chronic hypoxic pulmonary hypertension(HPH)rats and chronic hypoxic rats treated with novel KATP opener iptakalim.Methods sixty Sprague-Dawley(SD)male rats were randomly divided into control group, hypoxic group, low dose iptakalim group(0.75 mg·kg~(-1)·d~(-1)), and high dose iptakalim group(1.5 mg·kg~(-1)·d~(-1)).Except the first group, the other three groups were put into hypoxic and normobaric chamber (10%±0.5% O_2,8 h/day and 6 day/week) to establish chronic hypoxic model. After four weeks, the mean pulmonary arterial pressure(mPAP), RV/(LV+S)and the plasma concentration of NO were measured. RT-PCR was performed to analyze the mRNA expression of eNOS in pulmonary tissue. Western blot was performed to analyze the protein expression of eNOS, iNOS in pulmonary tissue. Results ① The level of mPAP and RV/ ( LV + S) were significantly higher in the hypoxic group than those in control group ( P < 0. 05 ) , Low dose iptakalim groupandhighdoseiptakalimgroupdecreased the level of mPAP and RV/( LV + S) significantly (P <0. 05). ② The level of NO was significantly lower in the hypoxic group than those in control group (P<0. 05). Low dose iptakalim group and high dose iptakalim group increased the level of NO significantly (P < 0. 05 ). ③ The mRNA and protein expression of eNOS in the hypoxic group were significantly lower than those in the control group (P < 0. 05 ). Low dose iptakalim group and high dose iptakalim group increased the expression of eNOS significantly ( P < 0. 05). High dose iptakalim group was more significant. Conclusion Pulmonary vascular endothelial dysfunction is induced by chronic hypoxia,and the level of NO, the mRNA and protein expression of eNOS are decreased. Iptakalim can improve the vascular endothelial dysfunction, increase the expression of eNOS and the level of NO and reverse hypoxic pulmonary hypertension.

Objective To present our experience with management of craniopharyngiomas by endoscopic endonasal approach Methods A retrospective review of clinical data of 65 patients who were treated for craniopharyngiomas by endoscopic endonasal approach from February 2012 to May 2016. All patients were analyzed by treatment effect, complications, and follow-up result. Results Total removal of the tumors were completed in 52 cases (80.0%), subtotal removal in 11 cases (16.9%), and partial resection in 2 cases (3.1%). The pituitary stalks were identified in 57 cases when surgery, and severed in 41 cases (71.9%). Postoperative visual acuity was improved in 31 cases (47.7%), and 6 cases remained in the preoperative level, whereas worsening occurred in 1 case. Worsening of the anterior pituitary function was reported in 21 cases (32.3%). Transient diabetes insipidus after operation was occurred in 45 patients (69.2%), and long-term diabetes insipidus was occurred in 9 cases (13.8%). Postoperative cerebrospinal fluid (CSF) leak was occurred in 4 cases (6.2%), accompanied with intracranial infection, and all these cases were repaired under endoscope again, 3 cases were saved, but 1 case was dead. Perioperative mortality rate was 4.6%. 52 patients were followed up for 4.0 ~ 45.0 (mean, 20.8) months, and 44 patients (84.6%) returned life to normal. Obesity developed in 8 patients (15.4%), with 2 recurrent cases and no deaths during follow-up period. Conclusion The endoscopic endonasal approach is a safe and effective minimally invasive surgery approach for treating craniopharyngiomas, and has its own unique advantage.

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

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.