ObjectiveTo observe the clinical efficacy ofShu Feng An Shen(expelling wind and calming mind) needling plus distal points selection in treating cluster headache.MethodForty patients with cluster headache were divided into two groups by using random number table, 20 cases in each group. The treatment group was intervened byShu Feng An Shenneedling plus distal points selection, while the control group was by acupuncture with conventional points selection. The pain intensity score (Visual Analogue Scale, VAS), lasting duration and attack frequency were observed.ResultThe VAS score, lasting duration and attack frequency were significantly reduced after treatment in the two groups (P<0.01), and the reduction was more significant in the treatment group (P<0.01).ConclusionShu Feng An Shenneedling plus distal points selection can produce a more significant efficacy in reducing pain intensity, lasting duration, and attack frequency in treating cluster headache compared with acupuncture with conventional points selection.

Objective: This work aimed to investigate the negative prognostic factors of bronchioloalveolar carcinoma (BAC) and adenocarcinoma with BAC characteristics, based on the 2004 pathological classification by the World Health Organization (WHO), which were further verified with the new pathological classification of lung adenocarcinoma (WHO 2011), to identify crucial factors that determine the prognosis of BAC and adenocarcinoma with BAC features, and to prove the coherence of the two pathological classi-fications in assessing clinical prognosis. Methods: Upon pathological diagnosis, some of the 193 cases of BAC or adenocarcinoma with BAC features were categorized into adenocarcinoma in situ (AIS) or minimally invasive adenocarcinoma (MIA), based on the 2011 WHO classification. Gender, age, tumor size, familial cancer history, smoking history, TNM stage, symptoms, duration of symp-toms, and the choice of treatment were recorded and analyzed for prognosis. The survival rate was calculated by Kaplan-Meier method. Log-rank test was introduced to compare the survival rate. Univariate and multivariate factors for the survival rate were analyzed by Cox proportional hazards regression model. Results:The overall 1-, 3-and 5-year survival rates were 84.3%, 60.6%, and 45.6%, respec-tively. Cox univariate analysis revealed that the tumor size, symptoms, TNM stage, pathological outcomes, and the choice of treatment were all prognostic factors. Cox multivariate analysis revealed that TNM stage was an independent prognostic factor for patients with BAC. Data from patients with AIS and MIA revealed better survival. Conclusion:The overall survival rate of BAC and adenocarcino-ma with BAC features are superior to that of other non-small cell lung cancer (NSCLC). The clinical symptoms are non-specific com-pared with other types of NSCLC. Clinical stage at diagnosis is a key prognostic factor, such that early correct diagnosis significantly improves survival. The new classification criteria of WHO, released in 2011, is more elaborate and more conducive to clinical practice.

Objective To investigate antimicrobial prophylaxis in clean operation,and provide evidence for formu-lating antimicrobial management measures.Methods 12 types of clean operation cases who discharged from a tertia-ry first-class hospital in May of 2011 -2014 were selected,120 cases were selected each year,patients were divided into group I and group II according to the types of operation,survey forms were designed,assessment criteria for the rationality of antimicrobial use was formulated, antimicrobial use in different years was compared. Results Prophylactic use of antimicrobial agents for clean operation decreased from 93.33% in 2011 to 35.00% in 2013 and 2014;rates of correct medication time increased from 43.75% in 2011 to 97.62% in 2013 and 92.86% in 2014 respectively;rates of medication according to indications,rational choice of antimicrobial agents,and rational treatment course increased from 42.86%,60.71 %,and 21 .43% in 2011 to 100.00% in 2014 (all P < 0.05 ). Conclusion Special rectification activities of antimicrobial use effectively promoted the standard application of anti-microbial agents for clean operation.

ObjectiveTo investigate the effect of recombinant human growth hormone (rhGH) on JAK2-STAT3 signaling pathway and on the growth of human colon cancer cells at different growth hormone receptor (GHR) expression status.MethodsLOVO and HCT-8 cell lines were selected after the colon cancer cells were screened using flow cytometry according to the GHR expression status.The growth of human colon cancer cells after intervention with rhGH was detected by MTT assay.The proliferation index ( PI),cell cycle,and apoptosis were detected by flow cytometry.The expression of JAK2-STAT3 signaling pathway proteins was detected by Western blot.ResultsHCT-8 cell line showed positive GHR expression (59.6％),and LOVO cell showed negative GHR expression (3.5％).The growth rate of HCT-8 cells increased after rhGH treatment.Compared with the untreated HCT-8 cells,the rhGH-treated HCT-8 cells showed reduced apoptosis,elevated PI,and increased G2/Mphase cells ( P =0.0073).Western blot revealed that rhGH up-regulated the proteins of pJAK2,pSTAT3,VEGF,Cyclin D1,and Bcl-xL in HCT-8 cells.In contrast,no such changes were observed in LOVO cells treated with rhGH.ConclusionsrhGH may promote the growth of HCT-8,the cell line of high GHR expression,and up-regulate the expression of a number of key nodes in JAK2-STAT3 signaling pathway.However,rhGH may not affect LOVO,the cell line of low GHR expression.

OBJECTIVETo investigate the ability of invasion and migration of breast cancer MDA-MB-231 cells under serum starvation and hypoxia, and the effect of antiangiogenic drugs, rh-endostatin and bevacizumab, on the ability of invasion and migration of breast cancer cells under serum starvation and/or hypoxia, in order to explore the potential risk of antiangiogenic therapy in clinics.

METHODSThe cells were randomized into 4 groups, i.e., group A: 10% fetal bovine serum (FBS) group; group B: hypoxia + 10% FBS group; group C: serum starvation group; group D: hypoxia + serum starvation group; each group was further divided into three subgroups as blank control, treated with rh-endostatin and bevacizumab, respectively. Cell counting kit-8 (CCK-8) was used to assess the inhibition rate of cell growth induced by endostatin and bevacizumab, in order to determine the proper working concentration and time of the two drugs. Transwell assay was conducted to detect the cell invasion and migration in vitro. The expressions of c-Met and MMP-9 were detected by Western blot. The cells treated with rh-endostatin or bevacizumab under serum starvation were tested by hybridization using Exiqon miBase 18.0 microarray. The miRNAs which exibited significant differences (P < 0.05) in miRNA hybridization were verified by real-time PCR assay.

RESULTSCCK-8 assay showed that the inhibition rates of MDA-MB-231 cells cultured with 800 mg/L rh-endostatin for 48 h and 24 h were (32.2 ± 2.5)% and (27.0 ± 1.3)%, respectively, showing a significant difference (P = 0.023). The inhibition rates of MDA-MB-231 cells cultured with 80 mg/L bevacizumab for 48 h and 24 h were (30.5 ± 1.4) % and (26.1 ± 2.4) %, respectively, showing also a significant difference (P = 0.015). The Transwell assay showed that in the starvation blank group, the number of invaded and penetrated cells were 28.8 ± 2.2 and 31.4 ± 1.5, respectively, significantly different from that in the rh-endostatin and bevacizumab groups (P < 0.05). The relative expressions of c-Met and MMP-9 were 0.213 ± 0.017 and 0.542 ± 0.048, respectively, with a significant difference from those of the groups treated with each drug (P < 0.05 for both). The numbers of penetrated cells in the Transwell assay treated with rh-endostatin in hypoxia were 17.5 ± 2.1 and 16.5 ± 2.8, respectively, and the numbers of penetrated cells in the Transwell assay treated with bevacizumab were 16.3 ± 3.5 and 17.5 ± 2.4, respectively, showing no significant difference among them (P > 0.05 for both). The ability of migration and invasion of MDA-MB-231 cells and the expression of c-Met and MMP-9 were not impacted by hypoxia (P > 0.05). Real-time PCR assay showed that only the levels of miR-2355 and miR375 were significantly and stably decreased in the cells which had increased ability of invasion and migration. The relative expression levels of miR375 and miR-2355 in the serum starvation blank group were 0.550 ± 0.036 and 0.852 ± 0.121, respectively, significantly lower than that in the groups treated with rh-endostatin or bevacizumab (P<0.05). In the serum starvation group, the expression levels of miR375 and miR-2355 of cells treated with rh-endostatin were 0.295 ± 0.012 and 0.253 ± 0.011, and the expression levels of cells treated with bevacizumab were 0.234 ± 0.020 and 0.309 ± 0.022, respectively, (P > 0.05 for all). Compared with the serum starvation blank group, the expression levels of miR2355 and miR375 were significantly decreased when cells were treated with rh-endostatin/bevacizumab under serum starvation, but no significant difference was found between the two drugs (P > 0.05). However, hypoxia did not affect the expressions of miR2355 and miR375 (P > 0.05).

CONCLUSIONSThe results of this study suggest that serum starvation can increase the ability of invasion and migration of breast cancer cells. Furthermore, both rh-endostatin and bevacizumab may enhance their invasion and penetration ability under serum starvation condition.

Angiogenesis Inhibitors ; adverse effects ; Bevacizumab ; adverse effects ; Breast Neoplasms ; pathology ; Cell Hypoxia ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Culture Media, Serum-Free ; Endostatins ; adverse effects ; Female ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; MicroRNAs ; analysis ; Neoplasm Invasiveness ; Proto-Oncogene Proteins c-met ; metabolism ; Random Allocation ; Real-Time Polymerase Chain Reaction ; Time Factors

Objective: This study aimed to examine the number of activated circulating endothelial cells (aCECs) in the peripheral blood of patients with non-small cell lung cancer (NSCLC), and investigate the relationship among aCECs, anti-angiogenic therapy, and prognosis of NSCLC patients. This study also aimed to identify novel predictive markers for anti-angiogenic therapy, and provide basic data and experimental basis for establishing an evaluation system for this therapy. Methods: A total of 142 NSCLC patients were randomly divided into the chemotherapy group (Group 1) and combined therapy group (i.e., chemotherapy plus endostatin, Group 2). The number of aCECs was measured using flow cytometry by detecting the expression status of CD105 and CD146 in the peripheral blood. The correlation between the changes in aCECs and efficacy of drug treatment was statistically analyzed using SPSS software. Results:The number of aCECs in Group 2 increased significantly at 8 and 29 d, two cycles, 50 and 71 d, and four cycles after treatment, respectively (P<0.05). In particular, aCECs amount in cases of progressive disease increased more significantly after combined therapy (P<0.05). A negative correlation was found between the treatment cycle and difference in aCECs amount before and after therapy (r=-0.970, P=0.001). A negative correlation was also observed between the difference in aCECs amount and time to tumor progression (TTP) (r=-0.351, P=0.039). Therefore, the difference in aCECs amount before and after therapy could serve as an important predictor for TTP in NSCLC patients. Conclusion:CD105 and CD146 reflected the activation status of endothelial cells, and responded to the drug treatment. Thus, CD105 and CD146 could act as ideal markers for aCECs. The number of aCECs increased during cancer progression, but significantly decreased after long-term treatment. Therefore, the change in aCECs amount may be a useful marker in predicting the efficacy of anti-angiogenic therapy.

Objectives To investigate the effect of IL-17 on proliferation and secretion of IL-1βin the human mesangial cell line (HMCL) and explore the molecular mechanism. Methods The proliferation of HMCL induced by IL-17 was detected by MTT, the secretion of IL-1βin HMCL was measured by ELISA and the expression of NF-κB was detected by immunocyto-chemistry. Results IL-17 did not impact HMCL proliferation (P>0.05) but dose-and time-dependently promoted the secretion of IL-1β(P<0.05). And the expression of NF-κB induced by IL-17 increased comparing to that in the control group (P<0.05). Con-clusions IL-17 may induce HMCL to secrete IL-1βvia NF-κB pathway that results in kidney injury.

Objective To evaluate cytological test of cerebrospinal fluid in the diagnosis of meningeal dissemination of tumor cells.Methods The clinical and imaging features of 85 cases with tumor cells diagnosed by thin-layer centrifugal cytological test of cerebrospinal fluid were retrospectively reviewed.The characteristics of cellular morphology and immunocytochemical staining were analyzed.Results The main presentations of all the patinets was meningeal irritation and neurological dysfunction.The features of the brain MRI were meningeal thicking and enhancement,intracranial abnormal signals and intracranial space occupying lesion in part of the patients.Atypical cells were found in 84 cases (98.8％) with the first sample test and immunocytochemical staining was conducted in 48 cases to identify the tissue origin.Meningeal carcinomatosis was shown to be the majority with lung cancer as the dominated tissue type and adenocarcinoma as the most common histological type.Others were lymphatic hematopoietic system (13 cases),melanomas (5 cases),primitive neuroectodermal tumor (3 cases) and glioma (1 case).In addition,12 cases were only proved to be cancer by cytological test of cerebrospinal fluid.Conclusion The thin-layer centrifugal cytological test of cerebrospinal fluid has a relatively high accuracy for detecting disseminated tumor cells of meninges and could be of great help to identify the source and type of lesion with immunocytochemical staining.

AIM:To investigate the regulation mechanism for insufficient KChIP 2 expression induces Ito,f downregulation and arrhythmogene-sis in cardiac hypertrophy .METHODS:Bidirectional manipulations of MG 53 expression were performed by adenoviral overexpression of MG53 or knockdown of MG53 with RNA interference in neonatal rat ventricular myocytes with or without PE stimulation .Ito,f was re-corded with patch clamp in whole-cell mode 48 h after adenoviral transfection .Then the WT or MG53 knockout ( MG53 -/-) mouse model of left ventricular hypertrophy induced by transverse aortic constriction ( TAC) were used to detect the susceptibility to ventricu-lar arrhythmia.RESULTS: Here, we show muscle-specific MG53 regulates KChIP2 expression and Ito,f densities, where they are downregulated in hearts from MG53 knockout mice and MG53 knockdown rat cardiomyocytes , but upregulated in MG53 overexpressed cells.MG53 expression is decreased in phenylephrine ( PE)-induced cardiomyocyte hypertrophy and restoration of MG 53 rescues PE-induced downregulation of KChIP2 and Ito,f.Furthermore, MG53 is decreased in a mouse model of hypertrophy induced by transverse aortic constriction and ablation of MG 53 increases the susceptibility to ventricular arrhythmia by exaggerating Ito,f remodeling.CON-CLUSION:These findings establish MG53 as a novel regulator of Ito,f and its central role in arrhythmogenesis in hypertrophy .

Objective To identify the histological features as well as factors influencing the course of HBeAg-negative chronic hepatitis B virus ( HBV)-infected patients with persistently normal alanine amino-transferase (ALT) levels (PNAL). Methods Ninety-eight HBeAg-negative chronic HBV-infected patients with PNAL who underwent percutaneous liver biopsy were recruited from October 2003 to March 2008. The ALT level, HBV markers, HBV DNA level and liver histological changes were detected. Comparison of means was done by t test and single factor analysis of variance. Nonparametric statistics was done by Marm-Whitey U test and Kruskal-Wallis test. Analysis of independent risk factor was done using Logistic model. The dianostic value of ALT level to significant liver histological changes was evaluated by receiver performance curve. Results Twenty-two point four percent and 17.3% of subjects had the histological activity index (HAI)≥4and fibrosis (F) score≥3 respectively. Subgroup analysis showed that subjects with ALT＞0.50 × upper limit of normal (ULN) had a significantly higher rate of HAI≥4 and F score≥3 than those with ALT≤0.50×ULN (HAI≥4:36.4% vs 11.1%, χ2 =8.881, P=0.003;F score≥3:27.3% vs 9.3%, χ2 =5.487, P= 0.019, respectively), and older subjects (more than 45 years old) had a higher proportion of HAI ≥4 than the younger (33.3% vs 13.4%, χ2 =4.923, P=0.027). Multivariate Logistic regression analysis revealed that a decade increase in age was the independent predictor of HAI≥4 (OR=2.410, P=0.023).Receive operating characteristic (ROC) curve showed that 87.0% and 90.7% of subjects with ALT＜0.50× ULN had histological changes of HAI＜4 and F score＜3 respectively. The proportions of HAI≥4 and F score≥3 in subjects with HBV DNA＜1×104 copy/mL were 14.9% and 12.8%, respectively. Conclusions Significant histological changes may be present in part of the subjects with persistently normal ALT and different HBV DNA levels, so that liver biopsy is very important, especially in those with age ＞45 years.Half time the ULN may serve as an appropriate cutoff value of normal ALT level for managing Chinese HBeAg-negative chronic HBV-int'ected patients.