The viewpoint of treating gallbladder and combinedtreatment of liver and gallbladder for liver diseasse isproposed and the therapeutic project of intravenousdrip of Mixture of Radix Bupleuri plus Radix SalviaeMiltiorrhizae is formulated.Sixty eases of chronic hep-atitis of damp—heat of liver—gallbladder with block-age of collaterals by stagnant blood were thus treated.Results revealed the relief of jaundice,decrease of en-zyme and inhibition of viral replication in treatinggroup are all better than the western drug controlgroup.

Objective To investigate the role of cell adhesion molecule L1 like (CALL) in the genesis and development of esophageal squamous cell carcinoma (ESCC).Methods From July 2007 to December 2010,a total of 100 patients with ESCC who received radical resection of esophageal cancer were enrolled.The ESCC tissues and corresponding tumor-adjacent normal tissues were obtained.The expression of CALl was determined by tissue microarray technology and immunohistochemical staining.The CALL over-expressed esophageal cancer cell line was established.The effects of CALL on cell migration and invasion were detected by wound-healing assay and Transwell assay,respectively.The effects of CALL on actin microfilament was analyzed by filamentous actin (F-actin) staining.Chi square test,Fisher's exact test,multivariate analysis and t test were performed for statistical analysis.Results The positive expression rate of CALL in ESCC tissues was 56 ％ (56/100),which was lower than that of tumor-adjacent normal tissues (95％,95/100),and the difference was statistically significant (x2=41.114,P＜0.01).There were statistically significant differences in CALL expression at protein level among patients with ESCC of different differentiation degree,different pathological T stage,lymph node metastasis and different TNM stage (x2=13.702,5.317,21.453,Fisher's exact test;all P＜ 0.05).The five year disease related survival rate of ESCC patients with down-regulated expression of CALL was 0(0/49),which was lower than those with normal CALL expression (25.5％,13/51),and the difference was statistically significant (x2 =43.338,P＜0.01).The median survival time of CALL expression down-regulated group was 17 months,and that of normal expressed group was 38 months.CALL expression was an independent risk factor of disease special survival rate (hazard ratio (HR) 0.353,95％ confidence interval (CI) 0.188 to 0.666,P=0.001).The results of wound-healing assay showed that the migration ability of CALL overexpressed CALL-k30 cells was lower than that of Vec-k30 cells in control group on 24 hours after wound.The results of Transwell invasion test showed the number of migrating cells penetrating CALL k30 cells attached to the inferior surface of the membrane was 44.000±13.748,which was less than that of the Vec k30 cells (154.333±25.007),and the difference was statistically significant (t=5.136,P=0.036).The results of F-actin staining demonstrated that actin filaments of CALL-k30 cells was 234.667 ± 65.118,which was lower than that of Vec-k30 cells (597.000± 119.929),and the difference was statistically significant (t=4.707,P=0.042).Conclusions CALL lowers the migration and invasion abilities of esophageal cancer cells by inhibiting F-actin microfilaments.Its abnormal expression may play an important role in the genesis,development and prognosis of ESCC.

Objective To investigate the effect of nerve cells apoptosis induced by endoplasmic reticulum stress (ERS) on brain injury after asphyxiating cardiac arrest and cardiopulmonary resuscitation (CA/CPR) in rats.Methods A total of 40 male Sprague-Dawley rats were randomly divided into control group and CA/CPR group (n=20).The CA/CPR models were established by asphyxia method/CPR.The levels of neuron specific enolization enzyme (NSE) and S100 beta protein (S100β protein) in serum at baseline and 0,3 and 6 h after CPR were detected by double antibody sandwich enzyme-linked immunosorbent assay (ELISA).The mRNA expressions of CCAAT-enhancer binding protein homologous protein (CHOP),activate transcription factor 4 (A TF4) and X-4 box binding protein 1 (XBP1) in the hippocampus were detected by reverse transcription-polymerase chain reaction (RT-PCR).The protein expressions of CHOP,B-cell lymphoma-2 (Bcl-2),Bcl-2 associated X protein (Bax) and cysteinyl aspartate specific proteinase 3 (caspase-3) in the hippocampus were measured by Western boltting.Neuronal apoptosis were detected by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL).Morphological and ultrastructural changes of the hippocampi were observed by light microscopy and electron microscopy.Results As compared to that in the control group,S100β protein in the CA/CPR group at 0,3 and 6 h after resuscitation was statistically different (P＜0.05);NSE protein level in the CA/CPR group at 6 h after resuscitation was significantly higher than that in the control group (P＜0.05).As compared with those in the control group,the mRNA expressions of CHOP,A TF4 and XBP-1 in the hippocampus of the CA/CPR group were obviously increased (P＜0.05).Significantly increased protein expressions of CHOP,Bax,and caspase-3,and statistically decreased Bcl-2 expression in the CA/CPR group were noted as compared with those in the control group (P＜0.05).The neuronal apoptosis rate in the CA/CPR group (29.74％±6.26％) was significantly higher than that in the control group (7.48％±4.34％,P＜0.05).The morphology changes and ultramicrostructure injuries of the hippocampus in the CA/CPR group were more obvious as compared with those in the control group.Conclusion CA/CPR in rats causes significant damage to brain tissues,and brain injury is aggravated gradually along with the prolongation of time,and the mechanism of brain injury may be connected with ERS-induced apoptosis of nerve cells.

Objective:To inhibit the Itch gene expression of T-lymphocytes and investigate the immune activity of T lymphocytes using ultrasound-targeted microbubble destruction (UTMD).Methods:T lymphocytes were separated by magnetic bead, and to establish an Itch gene targeted short hairpin RNA (shRNA) plasmid. There were three groups in this study: ①experimental group, Itch-shRNA plasmid-SonoVue microbubbles combined with ultrasound irradiation; ②control group, negative control shRNA plasmid-SonoVue microbubbles combined with ultrasound irradiation; ③blank group, untreated. Forty-eight hours after UTMD transfection, transfection efficiency was detected and analyzed by fluorescence microscopy and flow cytometry, the expression of Itch protein was measured with Western blotting.Seventy-two hours after UTMD transfection, the secretion of interleukin 2 (IL-2) and interferon γ (IFN-γ) in the cell supernatant were measured by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA was used to compare the differences between groups, and LSD- t test was used to compare the differences between groups. Results:The UTMD mediated shRNA transfection rate was 52.3%, and the relative expression levels of Itch protein in the experimental group, control group and blank group were 0.301±0.080, 0.773±0.101 and 0.719±0.090, respectively, and the difference was statistically significant( F=24.441, P<0.01). The Itch gene expression can be effectively suppressed in the experimental group. Seventy-two hours after transfection, the concentrations of IL-2 in the experimental group, control group and blank group were (417.3±37.1)ng/L, (158.7±17.3)ng/L and (147.0±10.2)ng/L, respectively, and the difference was statistically significant( F=118.701, P<0.001) and the concentrations of IFN-γ were (168.3±12.1)ng/L, (74.3±3.7)ng/L and (74.6±7.1)ng/L, respectively, and the difference was statistically significant ( F=126.833, P<0.001). The expression levels of IL-2 and IFN-γ in the experimental group were significantly higher than those in the control group and the blank group. Conclusions:UTMD mediated shRNA transfection can significantly decrease the expression of Itch and promote immune activity of T lymphocyte.

ObjectiveTo construct the rat model of high-normal blood pressure with excessive phlegm-dampness syndrome and analyze the serum metabolites and explain the metabolic characteristics of the model rats. MethodThe model rats of high-normal blood pressure with excessive phlegm-dampness syndrome were fed with high-fat diet and intraperitoneal injected with 7.625 mg·kg^-1 of Nω-nitro-L-arginine （L-NNA）， while rats in the normal group were fed chow diet and injected with the same amount of normal saline. The model was evaluated from the aspects of the general state， body weight， blood pressure and serum biochemical indexes， such as triglyceride （TG）， total cholesterol （TC）， high density lipoprotein cholesterol （HDL-C）， low density lipoprotein cholesterol （LDL-C）， blood glucose， malondialdehyde （MDA） and glutathione peroxidase （GSH-Px）. Liquid chromatography-mass spectrometry was used to analyze the changes of metabolites in rats serum. The conditions were as follows：mobile phase of 0.05% formic acid aqueous solution （A）-0.05% formic acid acetonitrile solution （B） for gradient elution （0-3 min， 2%B； 3-9 min， 2%-40%B； 9-18 min， 40%-98%B）， electrospray ionization （ESI）， positive and negative ion detection modes， acquisition range of m/z 80-1 000. Univariate and multivariate statistical analysis were used to screen the differential metabolites between groups which were used to metabolic pathways enrichment analysis. ResultCompared with the normal group， the rats in the model group had the characteristics of darker fur， lazy movement and loose stools， the body weight， blood glucose and contents of TG， TC， LDL-C， MDA increased significantly （P<0.05， P<0.01） and GSH-Px level decreased significantly （P<0.01）， the blood pressure remained stable at about 160/88 mmHg （1 mmHg≈0.133 kPa）. A total of 115 differential metabolites were screened in the positive and negative ion modes， of which 45 were up-regulated and 70 were down-regulated ［variable importance in the projection （VIP） value>1， P<0.05 and fold change （FC）≥2］， including indoleacetaldehyde， 5-hydroxytryptamine， lysophosphatidylcholine， phosphatidylcholine， xanthine and so on， which regulated blood pressure mainly through glycerophospholipid metabolism， linoleic acid metabolism， tryptophan metabolism and arachidonic acid metabolic pathways. ConclusionFeeding with high-fat diet and intraperitoneal injection of L-NNA can successfully construct the rat model of high-normal blood pressure with excessive phlegm-dampness syndrome， and indoleacetaldehyde， 5-hydroxytryptamine， lysophosphatidylcholine， phosphatidylcholine and xanthine can be used as characteristic serum differential metabolites of the rat model.

OBJECTIVE：To study the improvement effects of isopimpinelline on p-chlorophenylalanine（PCPA）-induced pineal injury model rats and its effect on expression of biological clock gene. METHODS ：Totally 60 rats were divided into blank control group（2％ polysorbate solution），model control group （2％ polysorbate solution），positive control group （melatonin，10 mg/kg） and isopimpinelline high-dose ，medium-dose and low-dose groups （3，1.5，0.75 mg/kg）. Except for blank control group ，rats in other groups were given PCPA intraperitoneally （450 mg/kg）to establish pineal injury model. After modeling finished ，they were given relevant medicine intragastrically ，once a day ，for consecutive 7 d. On the 6th day of administration ，the sleep latency and sleep duration of rats in each group were investigated by pentobarbital sodium coordination sleep test ；after last administration ， ELISA assay was used to determine the serum level of melatonin in rats. Fluorescence microscope and electron microscope were used to observe the pathological tissue and cell ultrastructure changes of the pineal gland. RT-qPCR was used to detect the mRNA expressions of biological clock gene Clock，Bmal1，Per1，Per2，Per3，Cry1，Cry2 in pineal gland of rats. RESULTS ：Compared with blank control group ，model control group had significantly longer sleep latency （P＜0.05）；serum melatonin ，mRNA expressions of Bmal1 and Per1 in pineal gland were significantly decreased （P＜0.05 or P＜0.01）while mRNA expression of Per3 was increased significantly （P＜0.05）. The pineal gland cell arrangement disorder ，nuclear pyknosis ，vacuolar degeneration increased and cell number decreased significantly ；mitochondria swollen ，cristae broken and pyknosis were observed. Compared with model control group ，the sleep latency of isopimpinelline high-dose group was shortened significantly （P＜0.05），sleep duration time was prolonged significantly （P＜0.05）；the levels of melatonin in serum ，mRNA expressions of Clock，Bmal1， Per1，Cry1 and Cry2 in pineal gland of rats were increased significantly （P＜0.05 or P＜0.01）. In isopimpinelline medium-dose group，the sleep latency was shortened significantly （P＜0.05）；the levels of melatonin in serum and mRNA expressions of Clock， Bmal1，Per1，Cry1，Cry2 in pineal gland were increased significantly （P＜0.05 or P＜0.01），while mRNA expression of Per3 was decreased significantly （P＜0.05）. In isopimpinelline low-dose group ，the levels of mRNA expressions of Clock，Bmal1，Per2 and Cry2 were increased significantly （P＜0.05），while mRNA expression of Per3 was decreased significantly （P＜0.05）. Cell arrangement disorder was improved and nuclear pyknosis vacuole degeneration was decreased to some extent in isopimpinelline groups；mitochondria swelled ，cristae fractured ，and pyknosis decreased to some extent. CONCLUSIONS ：Isopimpinelline can improve PCPA-induced pineal gland injury in rats ；it can up-regulate the expressions of positive regulators Clock，Bmal1 and negative regulators Per1，Per2，Cry1，Cry2，while down-regulate the expression of negative regulator Per3.

OBJECTIVETo observe the clinical efficacy differences between electroacupuncture (EA) and regular treatment for severe acute pancreatitis accompanied with paralytic ileus.

METHODSThis was a prospective pragmatic randomized controlled trial. A total of 140 cases of severe acute pancreatitis accompanied with paralytic ileus were randomly assigned into an EA group and a regular treatment group, 70 cases in each one. The patients in the regular treatment group were treated with regular treatment, including intensive care, gastrointestinal decompression, fasting, blood capacity supplement, acid suppression treatment, internal environment maintenance, infection prevention, inhibition of pancreatic exocrine secretion, etc. Based on the regular treatment, patients in the EA group were treated with EA at Zusanli (ST 36) and Zhigou (TE 6), 30 min for each treatment, once a day for totally 5 days. The VAS-based abdominal distension severity scale and abdominal pain severity scale were compared before and during treatment in the two groups, moreover, the number of patients who transferred to surgery or ICU was compared.

RESULTS(1) After the 1st EA, the abdominal pain and distension severity scales were both improved in the EA group, which were superior to those of the regular treatment group (all<0.05); afterwards, the abdominal distension and pain severity scales of each day in the EA group were all significantly superior to those of the regular treatment group (all<0.05). (2) The number of patients who transferred to surgery or ICU was not significantly different between the two groups (>0.05).

CONCLUSIONSEA at Zusanli (ST 36) and Zhigou (TE 6) can significantly reduce the abdominal distension and pain severity scales in patients of severe acute pancreatitis accompanied with paralytic ileus, indicating positive clinical significance; in addition, EA is safe and can be recommended to the treatment of severe acute pancreatitis in combination with treatment plan of integrated Chinese and western medicine.