AIM: To observe the function and morphological change of human umbilical vein endothelial cells(HUVECs) treated with lipopolysaccharide(LPS) and composite salviae dropping pills(DSP).METHODS: HUVECs were cultured and incubated within 10 mg/L LPS for 12 hours.Different final concentrations of composite salviae dropping pills(1 g/L,0.5 g/L,0.25 g/L,0.1 g/L) were added before and after LPS treatment. Cell viability,NO,NOS,ET-1 and intracellular calcium were measured.The cells were observed under inverted microscope,inverted phase contrast microscope,laser confocal scanning microscopy and transmission electron microscope.RESULTS: When given after LPS treatment,different final concentrations of composite salviae dropping pills played a protective role(P0.05).The HUVECs injured by LPS underwent apparent morphological change after treatment with composite salviae dropping pills.CONCLUSION: Composite salviae dropping pills have an evident protective effect on human umbilical vein endothelial cells exposed to LPS.

AIM: To observe the protective effect of composite salviae dropping pills against human umbilical vein endothelial cells(HUVECs) injure induced by hydrogen dioxide(H_2O_2) and discuss the mechanism of its therapeutic action on cardiac and encephalic diseases.METHODS: HUVECs were injured by 0.1 mmol/L H_2O_2 and then different final concentrations of composite salviae dropping pills(0.5 g/L,0.25 g/L,0.1 g/L) were added before and after the injury.Cell viability was measured by MTT assay.TBA method was used to detect the intracellular malonaldehyde. Nitric oxide(NO) in the culture medium was detected by using nitrate reductase assay,and immunocytochemisty was used to observe the expression of NOS2,NOS3 and NF-?B.Morphologic observation was also performed.RESULTS: HUVECs were injured by 0.1 mmol/L H_2O_2.Composite salviae dropping pills increased the cell viability apparently,inhibited the production of MDA induced by H_2O_2,regulated the generation of NO bilaterally and influenced the expression of NOS3 and NF-?B.CONCLUSIONS: Composite salviae dropping pills at concentrations of 0.5 g/L,0.25 g/L and 0.1 g/L protects HUVECs from injury by H_2O_2,no matter it is be added before or affter the injury.The possible mechanism is associated with its regulating the expression of NOS2,NOS3 and NF-?B.

Objective To investigate the relationship between acupuncture analgesia and cellular immunity, the acupuncture effect on expression of c fos mRNA, ppENKmRNA,iNOSmRNA and iNOS activity in the mouse peritoneal macrophage were studied. Methods Twenty BALB/c mice were randomly divided into 2 groups:a,the acupuncture group treated with 5Hz electroacupuncture (EA); b, the control group treated with no EA. The pain threshold was detected by K + ionophoresis method before and after EA or restraining. The experiment was performed by using in situ hybridization, NADPH NBT histochemistry, RNA dot blot and protein dot blot techniques. All dot blot signals were scanned for statistical analysis. Results The hybridization signals and iNOS activity appeared as granules in bluish violet color, localized in the cytoplasm of the macrophage. All dot blot signals were increased in the acupuncture group, when compared with those in the control group( P

In the present study the effects of electroacupuncture (EA) on substance P (SP), somatostatin (SOM) and serotonin (5-HT) immunoreactivity (IR) of APUD cells and nerve fibers in the pylorus and in the skin of "Zusanli" acupo- int of rats were studied using immunohistochemical technique. Forty rats were divided into 20 pairs, one of each pair for EA (bilateral "Zusanli" acupuncture for 20 min), the other one for control. After EA the pain threshold was mark- edly increased(P

In order to improve the technique for histochemical demonstration of the different lymphocyte patterns by alpha-naphthyl acetate esterase (ANAE) activity from a single microscopic slide, 10 kinds of comparative experiments were performed. The results showed as follows. The peripheral blood mononuclear cells first fixed in 0.4% glutaldehyde and postfixed in 15% formol calcium gave the best effect on preserving the enzyme activity and on connterstaning of the leucocyte nuclei. During formol calcium fixation, the erythrocytes were dissolved from the diluted blood film with PBS more easily than from the blood smear without any dilution. The lymphocytes were distributed more uniformly and densely in the former specimen than in the latter. When incubated with 0.5 mg alpha-naphthyl acetate/ml medium in hexoazotized pararosaniline for 1(1/2)～2 hrs at 37℃, the distinct ANAE patterns of circulating lymphocytes were more readily identified.In 28 healthy adults the differential counts of the lymphocyte ANAE patterns from the diluted blood specimen was similar to those found from the specimen of isolated lymphocytes. The focal pattern was about 56%, the diffuse pattern about 25% and the negative pattern about 19%; while the isolated lymphocyte specimen was compared with the diluted blood film from the same individuals, the focal pattern was decreased insignificantly, the diffuse pattern increased and the negative pattern decreased significantly.

Objective To explore the effects of vitamin C and niacinamide on the growth and differentiation of human primary cultured keratinocytes.Methods Normal human foreskin was used in this study.The epidermis was separated enzymatically from the dermis by thermolysin,and keratinocytes were isolated from the epidermis by digestion with trypsin plus EDTA.The single keratinocytes were cultured with undedying NIH-3T3 cells as feeder cells in a complete medium supplied with 50 mg/L (vitamin C group),niacinamide of 400 μmol/L(niacinamide group)or vehicle(control group).Immunocytochemistry and immunodot blot were performed using monoclonal antibodies directed against C-myc,cyclin D1,filaggrin and involucrin.Results The colony number was highest in vitamin C group,followed by the control group and niacinamide group,and the colony morphology in vitamin C group was similar to that in the control group,but distinct from that in the niacinamide group.A significant increase was noticed in the expression of C-myc,cyclin D1,filaggrin and involucrin in vitamin C-treated keratinocytes compared with the control keratinocytes(all P＜0.05);however,in niacinamide-treated keratinocytes,the expression of filaggrin was significantly enhanced(P＜0.01),that of involucrin remained unchanged(P＞0.05),while that of C-myc was depressed(P＜0.05).Conclusions These results demonstrate that vitamin C has a favorable effect on both the growth and differentiation of human keratinocytes,while niacinamide seems to only promote the differentiation but attenuate the growth of human keratinocytes.

OBJECTIVE To investigate the epidemiological characteristic of lower respiratory infection among the hospitalized patients in Tianlin Community Health Service Center so as to take effective measures to control the infection and reduce the emergence of drug resistant strains.METHODS Totally 6150 cases of patients discharged from our Center during from Jan 2004 to Sept 2006 were collected and among them 141 cases occurred hospital onset of lower respiratory infection and drug resistance was analyzed through contrasting.RESULTS The incidence rate of hospital onset of lower respiratory infection and drug resistance were 2.29% and 58.62%.Among the 141 cases,the incidence rate of ESBLs was 28.36% which mainly produced from Escherichia coli and Klebsiella pneumoniae.CONCLUSIONS Primary affection should actively treated and antibiotics should use rationally according to the drug sensitive cultivation to control the abuse of antibiotics and reduce the emergence of drug resistant strains.

The met-enkephalin (MEK), leu-enkephalin (LEK) and ?-endorphin (?-EP) immunoreactivities were detected in the circulating lymphocytes of 64 guinea pigs by PAP immunohistochemical method under the conditions of electroacupuncture, naloxone or exogenous opioid like peptides (OLP). Subsequently the ?-naphthyl acetate esterase (ANAE)was demonstrated on partial immunostained lymphocyte samples. The results showed that the OLP immunoreactivity with different intensity was found in approximate 96％ of the lymphocytes. There was a positive correlation between the ANAE-focal pattern lymphocytes and MEK-IIRL (intense immunoreactivity lymphocytes)or ?-EP-IIRL; whereas there was no significant correlation between the ANAE-focal pattern lymphocytes and LEK-IIRL. The MEK-IIRL, LEK-IIRL and ?-EP-IIRL were all increased markedly in the guinea pigs which underwent electroacupuncture at "Zusanli" points. There was no significant difference in MEK-IIRL, LEK-IIRL and ?-EP-IIRL, when the acupuncture group, naloxone group and control group were compared with each other. However, the LEK-IIRL, MEK-IIRL and ?-EP-IIRL could be depressed by naloxone, when the lymphocytes were preincubated with 10~(-7)mol/L of MEK, LEK or ?-EP in vitro. Hence, the naloxone group had a significant difference in MEK-IIRL, LEK-IIRL or ?-EP-IIRL when compared with the acupuncture group. A high peak of the lymphocyte MEK-IR among the range of 10~(-10)—10~(-8)mol/L of MEK preincubation in the acupuncture group when compared with the control group. It suggests that electroacupuncture may enhance activity of the unbound MEK receptor.

Capillary zone electrophoresis (CZE) was used to monitor the whole unfolding process of bovine insulin resulted from the dithiothreitol (DTT) reduction. An off-line matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) was employed to determine the molecular weight of the reaction products at the same time to confirm the observation results obtained by CZE. The structural change during the process of bovine insulin unfolding could be observed directly from the electropherogram and information of protein unfolding could be obtained simultaneously. The results indicated that as an effective tool of monitoring the conformational change of protein,the method of CZE was simple,quick,sensitive and lower sample consumption.

Objective: To observe the inhibitory effect of scopolamine(Spm) and chlopromazine (Clo) on withdrawal syndromes in morphine dependent rats. Methods: The intensity of withdrawal syndromes on the model of morphine dependent rats was recorded after single or muiltiple subcutaneous administration(sc) of Spm and Clo at different doses. Results: Withdrawal syndromes were markedly decreased when single Spm 1 mg/kg and Clo 0.5 mg/kg combined with morphine were injected (P<0.05). Spm+Clo(sc) had much stronger effects on inhibiting withdrawal syndromes after intraperitoneal (ip) naloxone in morphine dependent rats (P<0.01). Conclusion: Spm can act on Ach-receptor and relieve morphine withdrawal syndromes. Clo may have a synergistic action with Spm via α2-receptor in the locus coeruleus of the rat brain stem.