AIM: To investigate the effect of antisense oligodeoxynucleotides against platelet-derived growth factor B-chain(PDGF-B) in the proliferation of cultured rat hepatic stellate cells and collagen-Ⅰ production.METHODS: The phosphorothioate antisense oligodeoxynucleotides and missense oligodeoxynucleotides were synthesized.The rat hepatic stellate cells(HSCs) were treated with these oligodeoxynucleotides at different concentrations,respectively.The inhibition of the proliferating of cultured rat hepatic stellate cells was assayed by MTT method.The expressions of PDGF-B and collagenⅠ mRNA were detected by means of reverse transcription-polymerase chain reaction(RT-PCR).The expression of endogenous PDGF-B was determined by flow cytometry(FCM).Collagen-Ⅰin the supernatant was determined by ELISA.RESULTS: The results showed that the ASODN against the PDGF-B at a final concentration of 10 ?mol/L inhibited the HSCs proliferation,the expressions of PDGF-B and collagen-ⅠmRNA.FCM and ELISA demonstrated that the expression of endogenous PDGF-B and the production of collagen-Ⅰin HSCs were significantly lower than those in controls,whereas the controls(missense oligodeoxynucleotides) had no effect at the same concentration.CONCLUSION:PDGF-B ASODN inhibits the proliferation of cultured rat hepatic stellate cells,the endogenous expression of PDGF-B and the production of collagen.PDGF-B ASON might be useful for gene therapy in liver fibrosis.

Objective To evaluate the influences of the background music in operation room on the parturi-ents of first -time cesarean sections and the medical staff.Methods 204 parturients for elective cesarean sections were selected and divided into two groups according to the dates the operations:101 for the intervening group and 103 for the control group.The patients in the intervening group would enjoy both of the background music and language communication during C -section operation,while the cases in the control group would only have routine nursing.The self -evaluation of anxiety scale(SAS)and visual analogue scale(VAS)were applied to compare the degree of anxie-ty and pain during the operations.Meanwhile,the changes of averaged blood pressure and heart rate of the patients during the operations were also recorded and compared.A questionnaire survey was completed for all the medical staff who participated in the above operations.Results The SAS and VAS scores of the intervening group were (34.35 ± 7.40)points and (2.0 ±0.7)points,lower than those of the control group,which scored (47.85 ±5.58)points and (2.7 ±0.8)points.The differences between the two groups were significant(t =6.25,4.10,all P <0.05).while the averaged blood pressures and heart rates of the intervening group were (76.64 ±8.82)mmHg and (76.18 ± 12.09)times per minute respectively,which were also lower than those of the control group[(90.75 ±7.37)mmHg and (87.62 ±9.23)times per minute respectively],the differences between the two groups were significant(t =5.810,4.443,all P <0.05 ).All of the differences were statistically significant.75% of the medical staff thought background music was good for parturients and medical staff.Conclusion Background music in operation room can reduce the level of anxiety for the patients undergoing first -time cesarean section,and can reduce the degree of pain and discomfort of the patients,and also may stabilize the patients'blood pressures and heart rates,as well as regulate medical staff's mood,and improve their working efficiency.

Objective To investigate the effect of desflurane pretreatment on the density of muscarinic (M)-receptors on anoxic/reoxygenated rat cardiac myocytes. Methods Myocytes were obtained from ventricle of 2-3 day SD rats and incubated for 3 days. Then they were randomly allocated to one of 5 groups: group I received no treatment and served as control ( n = 5); group Ⅱ underwent 2 h hypoxia (liquid culture medium was insufflated with a gas mixture of 1 % CO2 , 1 % O2 and 98% N2) followed by 1h reoxygenation ( n = 6) ; group Ⅲ was pre-treated with 1.5 MAC desflurane (9% ) for 20 min, followed by 10 min washout before the same duration of hypoxia/reoxygenation as in group II ( n = 6) ; group IV underwent longer duration of hypoxia (48 h) followed by 24 reoxygenation ( n = 5) ; group V was pre-treated with 1.5 MAC desflurane (9% ) for 20 min followed by 10 min washout, then underwent longer duration of hypoxia/reoxygenation as in group IV ( n = 5) . The density of M-receptors was measured by the radio-ligand binding assay (RLBA) .Results The density of M-receptors was significantly lower in group Ⅱthan that in group I ( P

Purpose:To investigate the effect of in vitro antisense oligodeoxynucleotide of cyclin D 1 on the cyclin D 1 gene expression and cell proliferation of human stomach adenocarcinoma cell BGC 823 cell line.Methods:Phosphorothioate cyclin D 1 ASODN and random oligodeoxynucleotide (RODN) were synthesized and transfected into BGC 823 Cells. Their effects on cell proliferation were examined by MTT method,RT PCR method,immunohistochemical study.Results:Cyclin D 1 ASODN could significantly inhibit the growth of BGC 823 Cell lines.The RODN showed no such effect.The inhibition peaked at 48 hour after transfection by MTT method and was dose dependent.ASODN could downregulate the expression levels of cyclin D 1 mRNA and protein by RT PCR method and immunohistochemical study respectively.Conclusions:The data suggested that ASODN could specifically inhibit the expression of cyclin D 1 mRNA and protein and regulate cell cycle and cell proliferation of BGC 823 cells. [

BACKGROUND:It has been proved that bone marrow mononuclear cel transplantation can obviously improve neurological function of rats with cerebral hemorrhage. OBJECTIVE:To investigate the effects of transplanted bone marrow mononuclear cel s on the neurological function and apoptosis in perihematomal brain tissues fol owing cerebral hemorrhage in a rat model. METHODS:Twenty-four Sprague-Dawley rats were given stereotaxical injection of col agenase IV into the caudate nucleus to establish cerebral hemorrhage models in transplantation group (n=12) and model group (n=12), and then at 6 hours after cerebral hemorrhage, rats in these two groups were administrated 3x1010/L al ograft bone marrow mononuclear cel s and the same amount of PBS, respectively. Another 12 rats were given no interventions as control group. Neurological functions of rats were assessed at 1, 4, 8, 16 days after cerebral hemorrhage;pathological changes of the injury sites were observed at 16 days after transplantation;neuronal apoptosis rates in the perihematomal brain tissue were detected by flow cytometry at 2 and 4 days after transplantation. RESULTS AND CONCLUSION:The modified neurologic severity scores in the transplantation group were significantly lower than those in the model group at 8 and 16 days after cerebral hemorrhage (P<0.05). In the control group, cel s in each layer arranged closely with complete structure, and neurons and glial cel s were in good shape;in the model group, perihematomal brain tissues were loose with intercel ular gap, in which most neurons and glial cel s became necrotic;in the transplantation group, cel s in each layer arranged closely and regularly, and glial cel proliferation occurred. Besides, compared with the model group, the neuronal apoptosis rate in the transplantation group was significantly lower (P<0.05). To conclude, bone marrow mononuclear cel s can significantly enhance the neurological function recovery and reduce neuronal apoptosis in the brain of cerebral hemorrhage rats.

Objective To evaluate the effect and side effects of Kezhi capsule in short-term treatment of nonalcoholic fatty liver disease (NAFLD). Methods 60 NAFLD cases of outpatients according to the inclusion criteria of were randomly assigned to two groups: the treated group-30cases with the prescription of Kezhi capsule , and the control group-30cases with the prescription of Xuezhikang. The treatment Course of taking Kezhi capusule (1.25 g, tid, po) and Xuezhikang(0.6 g, bid, po)was 24 weeks. Results After treatment in both groups we saw the significant decrease of the levels of ALT, AST, TC, TG, LDL-C, CREAT, BUN, BMI and TCM Syndromes scores, and the improvement of the ultrasonographic findings of liver steatosis. In the Xuezhikang group we saw higher decrease of TC and TG than the Kezhi capsule group with statistic difference (P < 0.05), while in Kezhi capsule group we saw higher decrease of BMI , TCM Syndromes scores and the improvement of the ultrasonographic findings of liver steatosis than the xuezhikang group with statistic difference (P < 0.05). Conclusions The results show that kezhi capsule is effective for the treatment of NAFLD without obvious side effects.

Background and purpose: Hepatocellular carcinoma (HCC) is a hypervascular tumor associated with a poor prognosis and lack of effective treatments. Consequently, identifying novel therapeutic strategies are urgently needed. We have previously shown that the kringle 1 domain of human hepatocyte growth factor (HGFK1) is a more effective anti-angiogenesis molecule than angiostatin. In this study, we observed the effects and mechanisms of HGFK1 gene on the HCC. Methods: A recombinant adeno-associated vires carrying the HGFK1 gene (rAAV-HGFK1) was constructed.HCC of rat was induced by McA-RH7777. rAAV-HGFK1 was used to treat the rat, median survival time and metastasis rate were observed. Results: Ten days after tumor cell inoculation, surgery were performed to confirm the tumor formation, PBS, rAAV-EGFP or rAAV-HGFK1 was injected directly into the tumor nodule followed by portal vein injection. Results from our study demonstrated that rAAV-HGFK1 treatment significantly prolonged the median survival time of the HCC bearing rats from 30 days (PBS and rAAV-EGFP groups) to 49 days (rAAV-HGFK1 group). More importantly rAAV-HGFK1 inhibited tumor growth and completely prevented liver, lung and peritoneal metastasis. In the controlled PBS and AAV-EGFP group, liver and peritoneal metastasis rate were both 100%, and lung metastasis rate was 100% and 83%, respectively. While there was no metastasis found in treatment group, with only 33% of ascites happened. This was most possibly due to the primary tumor in liver but not due to the metastasis. Moreover, at a higher magnification (1000×), it was clear that the HGFK1 protein was expressed mainly in the cytoplasma of liver cells. In parallel, IHC staining of CD31 also demonstrated a significantly lower level of microvessel density (MVD) (6.21±1.6) in the liver tumor of the AAV-HGFK1 treatment group, as compared to the two control PBS and AAV-EGFP groups (25.1±2.1 and 26.8±2.5, respectively, P<0.01). HE staining showed that AAV-HGFK1 treatment induced large areas of necrosis in the tumor tissues, while minimal areas of necrosis were observed in the tumor tissue in the control groups. In addition, no toxicity appeared when high dosage (4.8× 1012 vg/rat) of rAAV-HGFK1 was administered in rats. Conclusion: Results from this study demonstrated that HGFK1 inhibited the growth and metastasis of HCC and prolonged the survival time of animals with HCC through anti-angiogenesis effects. No obvious toxicity was observed. It might be the novel promising treatment for HCC and other cancers.

AIM:To investigate the inhibitory effect of small interfering RNA ( siRNA) on the expression of protein kinase Cε( PKCε) in human hepatoma SK-Hep-1 cells, and the biological behaviors of the transduced cells , inclu-ding proliferation and invasion , were investigated.METHODS:The cultured SK-Hep-1 cells were divided into 3 groups, including PKCε-siRNA group , negative control ( NC)-siRNA group and control group .MTT assay was used to analyze the proliferation of the SK-Hep-1 cells in the respective groups , while invasion potency was determined by Transwell assay .The protein levels of functional biomarkers such as Ki 67 and matrix metalloproteinase 9 ( MMP-9 ) were measured by Western blotting .The Luciferase reporter gene assay was used to explore the activity of the NF-κB pathway .RESULTS:PKCεex-pression in SK-Hep-1 cells transfected with PKCε-siRNA was significantly down-regulated at both mRNA and protein levels compared with that in the normal SK-Hep-1 cells (P<0.01), with the decreases in the protein levels of Ki67 and MMP-9. The invasion and proliferation of SK-Hep-1 cells were obviously inhibited in PKCε-siRNA group compared with control group (P<0.01).Furthermore, the transcriptional activity of NF-κB was down-regulated when PKCε was effectively in-hibited by PKCε-siRNA (P<0.01).CONCLUSION:Down-regulation of PKCεinhibits the proliferation and invasion of hepatic carcinoma cells , which might be mediated via the NF-κB signaling pathway .

OBJECTIVE:To investigate the compatibility stability of Salvianolic acid for injection (SAFI) combined with 8 kinds of common solvents. METHODS:Referring to package inserts,SAFI was collected and combined with 0.9% Sodium chlo-ride injection,5% Glucose injection,10% Glucose injection,Compound sodium chloride injection,Mannitol injection,Glycerol fructose and sodium chloride injection,Sodium lactate Ringer's injection,Glucose and sodium chloride injection,250 mL each re-spectively,and then sealed in the dark at the room temperature. The appearance of mixtures were observed,pH value,the number of insoluble particles,maximal absorption wavelength and maximal absorbance were detected,and the contents of salvianolic acid B,rosmarinic acid,lithospermic acid and salvianolic acid Y in mixtures were determined by HPLC at 0,1,2,4,8 h after mix-ing. RESULTS:Under above condition,no obvious change was found in appearance or pH values of the mixtures within 8 h. Maxi-mal absorption wavelength ranged 284.5-286.0 nm. After mixed with Mannitol injection,the number of particles≥10μm(1-8 h af-ter mixing)and particles ≥25 μm(4-8 h after mixing)exceeded the scope of Chinese Pharmacopoeia(2015 edition);the maxi-mal absorbance changed significantly(RSD=9.17%,n=5);the relative content of salvianolic acid B,rosmarinic acid,lithosper-mic acid and salvianolic acid Y decreased by more than 10%(RSD=14.65%,6.45%,8.97%,12.49%,n=5);after mixed with Sodium lactate Ringer's injection,the relative content of rosmarinic acid and lithospermic acid changed greatly (RSD=14.57%, 7.28%,n=5);after mixed with 5% Glucose injection(4-8 h after mixing)and Glycerol fructose and sodium chloride injection(8 h after mixing),the relative content of rosmarinic acid were less than 90%(RSD=6.30%,4.86%,n=5);and the number of particles ≥25μm exceeded the scope of phamcopoeia after mixing with Glycerol fructose and sodium injection(0 h). The number of insoluble particles in other mixtures were in line with the standard of pharmacopoeia;maximal absorbance had no significant change(RSD<5%,n=5),and the relative content change of analytes were all less than 10%. CONCLUSIONS:Clinical appli-cation of SAFI combined with Mannitol injection,Sodium lactate Ringe's injection and Glycerol fructose and sodium injecrion should be avoided. After mixed with 5% Glucose injection,SAFI should be used within 4 h. SAFI can be compatible with 0.9%Sodium chloride injection,10% Glucose injection,Compound sodium chloride injection and Glucose and sodium chloride injec-tion.

The main ingredients of Ophiopogonjaponicas include steroidal saponins,flavonoids,polysaccharides,and so on,but the research on polysaccharides is relatively less.This particle will mainly review the chemical composition,analytical methods,and pharmacological effects of Ophiopogon polysaccharides.The Ophiopogon polysaccharides contain a variety of substances,such as MDG-1,Md-1,Md-2,OJP-1,etc;The main analysis methods were sulfuric acid method,phenol-sulfuric acid method,3,5-dinitrosalicylic acid (DNS) colorimetric method,and near-infrared spectroscopy combined with partial least squares regression method.Ophiopogon polysaccharides can effectively improve the cardiovascular system diseases,with antihypoxia,anti-inflammatory,antitumor,anti-oxidation,and other pharmacological effects.