Objective To observe the distribution of TCM syndromes in patients with functional constipation (FC). Methods One hundred and five cases of patients with FC were analyzed with relevant factors and summarized to syndromes of FC by cluster analysis. Results There was a correlation between distribution of syndromes of FC and patient's age, job occupation, course of disease, incentives and patient's eating habits. The distribution of syndromes analyzed by cluster analysis was consistent with diagnostic criteria. Conclusion The syndromes of FC are divided into heat constipation, qi constipation, deficiency constipation and cold constipation. Among these syndromes, the number of excess constipation patients especially heat constipation is more than the number of others.

Objective To investigate the effects of preconditioning with different concentrations of nerve growth factor (NGF)-β on ischemia-reperfusion (I/R) injury in isolated rat hearts.Methods Thirty-two male Sprague-Dawley rats weighing 200-300 g,aged 8-10 weeks,were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg and heparin 500 IU/kg.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution oxygenated with 95％ O2-5％ CO2 at 37 ℃.After 10 min of stabilization,the isolated hearts were randomly divided into 4 groups (n=8 each): I/R group and different concentrations of NGF-β groups (groups N1-N3).In group I/R,the hearts were continuously perfused with K-H solution for 30 min,perfusion was suspended for 30 min followed by 120 min reperfusion.In groups N1-N3,the hearts were continuously perfused with K-H solution containing NGF-β0.1,0.2 and 0.4 ng/ml,respectively,for 20 min,followed by 10 min washout,and perfusion was suspended for 30 min followed by 120 min reperfusion.HR,Left ventricular end-diastolic pressure (LV-EDP),left ventricular developed pressure (LVDP) and + dp/dtmax were measured at the end of 10 min stabilization (baseline,T1),immediately before suspension of perfusion (T2),and at 5,30,60 and 120 min of reperfusion (T3-6).The activities of creatine kinase MB (CK-MB) and lactate dehydrogenase (LDH) in the coronary effluent were measured at T1 and T3-T6.Myocardial tissues were obtained at T6 for detection of apoptosis (by TUNEL) and for microscopic examination.Apoptotic index (AI) was calculated.Results Compared with the baseline value at T1,LVDP and + dp/dtmax were significantly decreased,and LVEDP and activities of LDH and CK-MB were significantly increased at T3-T6 in each group,HR was significantly decreased in groups I/R,N2 and N3,while HR was significantly increased in group N1 (P ＜ 0.05).Compared with group I/R,LVDP,+ dp/dtmax and HR were significantly increased,LVEDP,activities of LDH and CK-MB and AI were significantly decreased in group N1,LVDP,+ dp/dtmax and HR were significantly decreased,and activities of LDH and CK-MB were significantly increased in group N2,and LVDP,+ dp/dtmax and HR were significantly decreased,and LVEDP,activities of LDH and CK-MB and Al were significantly increased in group N3 (P ＜ 0.05).LVDP,+ dp/dtmax and HR were significantly lower,and LVEDP,activities of LDH and CK-MB and AI were significantly higher in groups N2 and N3 than in group N1,and in group N3 than in group N2 (P ＜ 0.05).The pathologic changes were reduced in group N1 compared with I/R,N2 and N3 groups.Conclusion Preconditioning with the optimum concentration of NGF-β can at-tenuate I/R injury in isolated rat hearts,however,the injury can be aggravated when the concentration is too large,and inhibition of apoptosis in myocardial cells is involved in the mechanism of myocardial protection.

Objective To evaluate the effect of pretreatment with nerve growth factor-beta (NGF-β) on ischemia-reperfusion (I/R) injury in isolated rat hearts by comparing it with NGF-β preconditioning.Methods Pathogen-free male Sprague-Dawley rats,weighing 200-300 g,aged 8-10 weeks,were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with KH solution aerated with 95％ O2 and 5％ CO2 at 37℃.Twenty-four isolated rat hearts were randomly divided into 3 groups (n =8 each) using a random number table:I/R group,NGF-β pretreatment group (group NGFPt) and NGF-β preconditioning group (group NGFPc).The hearts were perfused with K-H solution for 10 min (equilibration) in each group.In group I/R,the hearts were continuously perfused with K-H solution for 30 min.The hearts were continuously perfused with K-H solution containing NGF 0.1 μg/ml for 20 min before ischemia in group NGF-Pt.The hearts were continuously perfused with K-H solution containing NGF 0.1 μg/ml for 20 min followed by 10 min washout before ischemia in group NGFPc.The perfusion was suspended for 30 min followed by 120 min of reperfusion with K-H solution in each group.HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and + dp/dtmax were measured at the end of 10 min equilibration (baseline,T1),immediately before suspension of perfusion (T2),and at 5,30,60 and 120 min of reperfusion (T3-6).The activities of creatine kinase isoenzyme-MB (CK-MB) and lactate dehydrogenase (LDH) in coronary effluent were measured at T1 and T3-6.Myocardial specimens were obtained at T6 for detection of myocardial apoptosis (by TUNEL) and for microscopic examination.Apoptotic index (AI) was calculated.Results Compared with the baseline value at T1,+ dp/dtmax was significantly decreased,and LVEDP and activities of CK-MB and LDH were increased at T3-6 in each group,LVDP and HR were decreased at T3-6 in group I/R,LVDP was decreased at T3,4 in group NGFPt and at T3-6 in group NGFPc,and HR was increased at T2-6 in NGFPt and NGF Pc groups (P ＜ 0.05).Compared with group I/R,LVDP,+ dp/dtmax and HR were significantly increased and LVEDP and activities of CK-MB and LDH and AI were decreased in NGFPt and NGFPc groups (P ＜ 0.05).Compared with group NGFPc,LVDP,+ dp/dtmax and HR were significantly increased,while the LDH activity and AI were decreased (P ＜ 0.05) and no significant changes were found in LVEDP and CK-MB activity in group NGFPt (P ＞ 0.05).The pathologic changes of myocardium were significantly reduced in NGFPt and NGFPc groups as compared with I/R group.Conclusion Pretreatment with 0.1 μg/ml NGF-β attenuates I/R injury in isloated rat hearts,and the efficacy is superior to that of NGF-β preconditioning.

Acupuncture clinical trials are designed to provide reliable evidence of clinical efficacy, and SCI papers is one of the high-quality clinical efficacy of acupuncture research. To analyze these papers published in high impact factor journals on acupuncture clinical trials, we can study clinical trials from design to implementation, the efficacy of prevention and cure, combined with international standard practices to evaluate the effectiveness and safety of acupuncture. That is the core of acupuncture clinical trials, as well as a prerequisite for outstanding academic output. A scientific and complete acupuncture clinical trial should be topically novel, designed innovative, logically clear, linguistically refining, and the most important point lies in a great discovery and solving the pragmatic problem. All of these are critical points of papers to be published in high impact factor journal, and directly affect international evaluation and promotion of acupuncture.
Acupuncture ; Acupuncture Therapy ; Clinical Trials as Topic ; Humans ; Journal Impact Factor ; Research Design

AIM: Some studies found that the glucose uptake ability of diabetes mellitus mice is stronger after feeding with taurine. Whether the taurine can affect adipocytes or not deserves further studies. This article investigates the effects of taurine on the differentiation of 3T3-L1 preadipocytes and detects the mechanics. METHODS: Experiments were performed in the Laboratory Center of Xiangya Third Hospital of Central South University from July to September 2006. ①3T3L1 preadipocytes were provided by Shanghai Cell Bank of China Academy of Sciences. ②3T3-L1 preadipocytes were cultured in high glucose DMEM medium containing fetal bovine serum of 0.10 volume fraction, 108 u/L benzylpenicillin and 80?1010 U/L streptaquaine. After confluence, cells at 5?107 L-1 were incubated in culture flask, and induced with 0.5 mmol/L IBMX, 0.5 mg/L insulin and 1 ?mol/L dexamethasone. The cells in an experimental group were intervened with taurine, whereas these in a control group were not treated with taurine. Oil O staining was used to observe the development of adipose cells. C3T3-L1 preadipocytes were treated with 10,20 mmol/L taurine respectively for 24 and 48 hours. RNA and protein were extracted in the control group and the experimental group. ③Genes related to adipose cells development was examined by RT-PCR and Western-blot. RESULTS: ①The number of cells stained by oil O was less in the experimental group than the control group in 3T3-L1 preadipocytes treated with 10 mmol/L taurine for 14 days. ②After C3T3-L1 preadipocytes were treated with 10,20 mmol/L taurine respectively for 24 and 48 hours, there were no changes in expression of Insig-2 protein, PPAR, Insing-2, adiponectin, adiponectin receptor, GLUT-4, AP-2 mRNA. CONCLUSION: Taurine inhibits the differentiation of 3T3-L1 preadipocytes, but the mechanics still need to be investigated.

Objective To evaluate the role of 1-phosphatidylinositol 3-kinase (PI3K) signaling pathway in nerve growth factor-beta (NGF-β)-produced mitigation of cell apoptosis induced by endoplasmic reticulum stress during hypoxia-reoxygenation (H/R) in rat cardiomyocytes.Methods H9c2 cells were seeded in 96-well plates at a density of 5× 105 cells/ml (100 μl/well).The wells were randomly divided into 5 groups (n =15 each) using a random number table: control group (group C);group H/R;NGF-β group (group N);NGF-β+NGF receptor trkA antagonist K252a group (group N+K);NGF-β+ PI3K inhibitor LY294002 group (group N+L).The cells were exposed to 95％ N2-5％ CO2 for 4 h in an anaerobic incubator, followed by reoxygenation in a standard incubator for 4 h in H/R, N, N+K and N+L groups.In addition, the cells in N, N + K and N + L groups were incubated in a standard incubator containing NGF-β, the mixture of NGF-β and K252a and the mixture of NGF-β and LY294002, respectively, during reoxygenation, and the final concentrations of NGF-β , K252a and LY294002 were 50 ng/ml, 100 nmol/L and 50 μmol/L, respectively.The cell viability was detected by using CCK-8 assay, and the cell survival rate was calculated.The cell apoptosis was examined by flow cytometry, and apoptosis rate was calculated.The expression of glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), caspase-12, Akt and phosphorylated Akt (p-Akt) was detected by Western blot.The ratio of p-Akt to Akt was calculated.Results Compared with group C, the cell survival rate was significantly decreased, and the apoptosis rate was increased in H/R and N groups, and the expression of GRP78, CHOP and caspase-12 was significantly up-regulated in group H/R, and p-Akt/Akt was significantly increased in group N (P＜0.05).Compared with group H/R, the cell survival rate was significantly increased, the apoptosis rate was decreased, the expression of GRP78, CHOP and caspase-12 was up-regulated, and p-Akt/Akt was increased in group N (P＜0.05), and no significant change was found in the parameters mentioned above in N+K and N+L groups (P＞0.05).Compared with group N, the cell survival rate was significantly decreased, the apoptosis rate was increased, the expression of GRP78, CHOP and caspase-12 was up-regulated, and p-Akt/Akt was decreased in N+K and N+L groups (P＜0.05).Conclusion NGF-β can mitigate the cell apoptosis induced by endoplasmic reticulum stress during H/R, and activation of PI3K signaling pathway is involved in the mechanism.

Pancreatic neuroendocrine tumors (P-NETs) are a group of heterogeneous tumors,including functional and nonfunctional ones.With the enhancement of clinicians' awareness about this disease and the improvement of imaging diagnostic techniques,the incidence of P-NETs has obviously increased in the past years.Based on the mitotic counting and Ki-67 positive index,the grading classification is of great value for the diagnosis,treatment and even prognosis of P-NETs.P-NETs are a group of malignant tumors with inert biological behaviors,whose surgical resection rate and long-term survival is much better than those of pancreatic ductal adenocarcinoma.P-NETs have different malignant potentials.Clinicians need to develop a comprehensive treatment plan in combination with the patient's symptoms,tumor grading classification and TNM staging information.Surgery is the only curable way to cure P-NETs.Even if radical resection is not suitable,palliative surgery may alleviate the patients,symptoms,and even prolong their survival time.According to the tumor location,size,quantity,degree of grading,local invasion and distant metastasis,different surgical procedures should be selected.

Objective To estimate the biological effects of leptin on human HaCaT keratinocytes and explore their molecular mechanisms.Methods Cell counting kit-8 (CCK-8) was used to evaluate the proliferation of cultured HaCaT cells treated with different concentrations of leptin for 24 and 48 hours.Some HaCaT cells were classified into four groups to remain untreated,be treated with leptin (100 μg/L) and piceatannol (a specific inhibitor of STAT3 phosphorylation) alone or in combination for 24 hours,respectively,followed by the evaluation of cell proliferation using CCK-8 kit.Flow cytometry was performed to assess cell cycle of HaCaT cells treated with leptin of 100 μg/L,Western blot to determine the phosphorylation level of Erk1/2 and STAT3 in HaCaT cells treated with leptin of 100 μg/L for different durations.Statistical analysis was done by Student's t-test for unpaired data using GraphPad Prism 5 software.Results The proliferation of HaCaT cells was accelerated to different degrees after treatment with leptin of 50 and 100 μg/L for 24 and 48 hours,and the accelerating effect was in a dose-dependent manner within 24 hours (r =0.9989,P ＜ 0.05).Piceatannol apparently inhibited the promotive effect of leptin on the proliferation of HaCaT cells.There was an obvious elevation in the percentage of cells at S phase ((57.70 ± 5.88)％ vs.(42.50 ± 7.55)％,P ＞ 0.05),but a significant decrease in that at G0/G1 phase ((39.70 ± 1.57)％ vs.(45.20 ± 1.44)％,P ＜ 0.05),with a significant increase in proliferation index (0.603 ±0.0157 vs.0.564 ± 0.0144,P ＜ 0.05) in HaCaT cells treated with leptin of 100 μg/L for 24 hours compared with the untreated controls.Western blot showed that leptin of 100 μg/L markedly enhanced the phosphorylation level of STAT3 in HaCaT cells.Conclusion Leptin may upregulate the proliferation of HaCaT cells through activation of STAT3 pathway.

Objective To evaluate the effects of nerve growth factor-beta (NGF-β) pretreatment on cell apoptosis induced by endoplasmic reticulum stress during ischemia-reperfusion in isolated rat hearts.Methods Male Sprague-Dawley rats,weighing 180-220 g,were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.The hearts were excised and perfused in a Langendorff apparatus with K-H solution aerated with 95％ O2 and 5％ CO2 at 37 ℃.Thirty-two isolated rat hearts were randomly divided into 4 groups (n =8 each) using a random number table:control group (group C),I/R group,NGF-β pretreatment group (group N) and NGF-β combined with K252a (trkA receptor antagonist) pretreatment group (group N + K).In group C,the hearts were continuously perfused with K-H solution for 195 min.The hearts were perfused with K-H solution for 45 min in group I/R.In N and N + K groups,the hearts were perfused with K-H solution for 15 min,and then with K-H solution containing 0.1 μg/ml NGF-β and 0.1 μg/ml NGF-β mixed with 100 nmol/L K252a,respectively,for 30 min.The perfusion was suspended for 30 min followed by 120 min of reperfusion with K-H solution in I/R,N and N + K groups.HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and + dp/dtmax were measured at the end of 15 min equilibration (baseline) and at 5,30,60 and 120 min of reperfusion.Myocardial specimens were obtained at 120 min of reperfusion for detection of myocardial apoptosis (by TUNEL) and expression of glucose-related protein 78 (GRP78),CCAAT/enhancer-binding protein homologous protein (CHOP),and caspase-12 (by Western blot analysis).Apoptosis index was calculated.Results Compared with group C,HR,LVDP and + dp/dtmax were significantly decreased,and LVEDP,apoptosis index and expression of GRP78 and CHOP were increased in I/R and N groups,and the expression of caspase-12 was upregulated in I/R group.Compared with group I/R,HR,LVDP,and + dp/dtmax were significantly increased,and LVEDP,apoptosis index and expression of GRP78,CHOP and caspase-12 were decreased in group N,and the expression of GRP78 was down-regulated in group N + K.There was no significant difference in cardiac function indexes between group I/R and N + K.Compared with group N,HR,LVDP and + dp/dtmax were significantly decreased,and LVEDP,apoptosis index,and expression of GRP78,CHOP and caspase-12 were increased in group N + K.Conclusion NGF-β pretreatment can protect the isolated rat hearts against ischemia-reperfusion injury,and inhibition of the endoplasmic reticulum stress-triggered cell apoptosis after activating trkA receptors is involved in the mechanism.

Objective To investigate the effect of propofol anesthesia on electroconvulsive therapy (ECT)-induced hyperphosphorylation of Tau protein in hippocampus in depressed rats.Methods Thirty-two female WYK rats in which the total score was 30-120 after Open-field test,aged 24 weeks,weighing 200-250 g,were randomly divided into 4 groups ( n =8 each):control group (group C),propofol group (group P),ECT group (group E)and propofol + ECT group (group PE).In groups C and E,the animals received intraperitoneal normal saline 5 ml,and in addition the animals received ECT 15 min later in group E.In groups P and PE,the animals received intraperitoneal 100 mg/kg propofol 5 ml,and in addition the animals received ECT 15 min later in group PE.The learning and memory function was assessed by Morris water maze test at 24 h after ECT.The animals were sacririced at 6 h after Morris water maze test and the hippocampal tissues were removed for determination of the expression of phosphorylated Tau protein.Results Compared with group C,the escape latency was significantly prolonged,the swimming time was significantly shortened in groups P,E and PE,the expression of phosphorylated Tau protein in hippocampus was down-regulated in group P,and the expression of phosphorylated Tau protein in hippocampus was up-regulated in group E ( P ＜ 0.05).Compared with group E,the escape latency was significantly shortened,the swimming time was significantly prolonged,and the expression of phosphorylated Tau protein in hippocampus was down-regulated in group PE (P ＜0.05).Conclusion The mechanism by which propofol anesthesia improves cognitive impairment induced by ECT may be related to inhibition of hyperphosphorylation of Tau protein in hippocampus in depressed rats.