Objective To explore the effects of low molecular weight heparin (LMWH) on the virological and pathological changes of newborn mouse liver congenitally infected with human cytomegalovirus (HCMV).Methods Sixty healthy pure line clean level BALB/c mice which were about 10 weeks old (half were female) were divided into five groups (six pairs in each group).The mice in LMWH intervention group and positive control group were intraperitoneally inoculated with 6.0 lg tissue culture infective dose50 (TCID50) of HCMV AD169; those in blank control group were intraperitoneally injected with 0.5 mL dulbecco's modified Eagles medium (DMEM) ; then all the mice were paired to mate.The pregnant mice in LMWH intervention group Ⅰ were subcutaneously injected with LMWH 1000 U/kg daily for 10 days; those in group Ⅱ were subcutaneously injected with LMWH 1000 U/kg daily for 5 days and their newborn mice were subcutaneously injected with LMWH 1000 U/kg daily for 5 days; those in group Ⅲ were subcutaneously injected with LMWH 1000 U/kg daily for 10 days in their newborn mice.All these newborn mice were sacrificed at day 10 of birth.The liver was removed for virus isolation,dry-wet weight determination,pathology examination and quantitative fluorescence-polymerase chain reaction (PCR) detection.The comparison among groups was done by analysis of variance.Results HCMV was isolated from the supernatant of liver tissue homogenate in 10-day positive control newborn mice,while HCMV was isolated in 24-day newborn mice of the other three groups of LMWH intervention.Pathology confirmed that positive control liver tissue had inflammatory changed,liver cell inflammatory swelling degeneration,vacuoles degeneration,specific HCMV inclusion body in nuclear,and portion of liver cell necrosis,while liver pathological results of LMWH intervention group showed mild liver cell inflammatory changes and slight cell inflammatory swelling degeneration,which were similar to the blank control group.The moisture of liver tissue contents in LMWH intervention group decreased more obviously than positive control group.The HCMV DNA loads in 50 mg liver tissues of LMWH intervention groups Ⅰ,Ⅱand Ⅲ were (3.26±0.43),(3.26±0.41) and (3.32±0.51) lg copy,respectively,which were significantly lower than that of positive control group [(7.38 ± 0.53) lg copy; F =314.620,P0.01],while there were no significant differences among LMWH intervention groups (P＞0.05).Conclusion LMWH intrauterine and postnatal interventions can significantly reduce HCMV DNA replication in hepatocytes,and relieve inflammatory changes in liver tissue.

Objective To discuss the validity and the feasibility of modified specific immunotherapy of hypoglossis allergen in the treatment of children allergic asthma. Methods 100 children with allergic asthma were selected from October,2007 to April ,2008 in our hospital and divided into the observation group and the control group. The control group adopted routine method, the observation group made modifi-cation based upon routine nursing, placing emphasis on intervention of cognition and behavior of children and their parents, improvement of treatment compliance, whole- process, dynamic and continuous observa-tion of the treatment process, making individualized health education plan. The treatment compliance, total score of asthma control, and pulmonary function examination of impulse oscillation(IOS) were compared be-tween the two groups. Results The observation group was superior to the control group in treatment com-pliance, pulmonary function examination and the control results of asthma. Conclusions Specific im-munotherapy of hypoglossis allergen combined with modified nursing method can increase treatment com-pliance of children and lead to better results.

Objective To study the effects of rehabilitation interventions on the earthquake injured to reduce the incidences of disability,to improve the recovery of overall function of the wounded in the earthquake and to guarantee their return to family and society.Methods The rehabilitation team·work provided one-month comprehensive intervention on 92 fracture patients after earthquake and assessed with scale of participate and activities ability,before,after and in the middle of intervention.Results The ability of understanding and communication,moving body,taking care of oneself,getting along with others,undertaking daily activities.social participating and the scores from the scale of participate and activities ability improved significantly(P<0.05).Conclusion Rehabilitation interventions with team-work can effectively enhance the abilities of the earthquake injured to participate in activities and improve quality of life.

BACKGROUND:Myeloid cell (TREM-1) is an important mediator of the signal transduction pathway in inflammatory response. In this study, a mouse model of acute lung injury (ALI) by intraperitoneal injection of lipopolysaccharide (LPS) was established to observe the expression pattern of TREM-1 in lung tissue and the role of TREM-1 in pulmonary inflammatory response to ALI. METHODS:Thirty BALB/C mice were randomly divided into a normal control group (n=6) and an ALI group (n=24). The model of ALI was made by intraperitonal injection of LPS in dose of 10 mg/kg. Specimens from peripheral blood and lung tissue were collected 6, 12, 24 and 48 hours after LPS injection. RT-PCR was used to detect TREM-1 mRNA, and ELISA was employed for detection of TREM-1 protein and TNF-a protein, and HE staining was performed for the pathological Smith lung scoring under a light microscope. RESULTS:The expressions of TREM-1 mRNA in lung tissue and blood of the ALI group 6, 12, 24, and 48 hours after injection of LPS were higher than those in the control group. The levels of TREM-1 protein and the levels of TNF-a protein in lung tissue of the ALI group 6, 12, 24, and 48 hours after LPS injection were higher than those of the control group; the level of TREM-1 protein peaked 12 hours after LPS injection, but it was not significantly correlated with the expression of TREM-1 mRNA (P=0.14); the TNF-a concentration was positively correlated with TREM-1 levels in lung tissue and with Smith pathological score (r=0.795, P=0.001:r=0.499, P=0.034), but not with the expression of TREM-1 mRNA (P=0.176). CONCLUSIONS:The expression of TREM-1 mRNA in lung tissue of mice with ALI is elevated, and the expression of TREM-1 mRNA is related to the level of TNF-a and the severity of inflammatory response to ALI. The expressions of the TREM-1 gene are not consistent with the levels of TREM-1 protein, suggesting a new functional protein involved in immune regulation.

Objective To investigate the role of CD4+ CD25+ regulatory T lymphocyte(Tregs)in the immunological pathogenesis of liver fibrosis.Methods Twenty-six rats were divided into two groups:control group(6 rats)and model group(twenty rats).The rat model of liver fibrosis was induced bv subcutaneous injection of carbon tetrachloride(CCl4).The serums were collected for detection of hepatic function and fibrosis parameters.Hepatic tissue samples were used to observe the histopathological changes.The flow cytometry was used to detect the proportions of Tregs in both peripheral blood and spleen.The data were evaluated by t-test.The relationship between two variables was analyzed using Pearson linear correlation.Results The levels of serum alanine aminotransferase (ALT)and aspartate aminotransferase (AST) were significantly increased,but the level of serum albumin (Alb) was obviously decreased.The concentrations of serum hyaluronic acid (HA),procollagen type Ⅲ(PCⅢ),collagen type Ⅳ(CⅣ)of the model group increased to(177.42±61.25)μg/L,(34.86±7.47)μg/L and(7.32±3.71)μg/L,respectively,which were higher than those in the control group(t=-3.670,-5.661,-3.950,respectively;all P<0.01).In model group,hepatic lobules were full of collagen fibers and the hepatic pseudolobule formation was observed.The proportion of peripheral blood Tregs in CD4+ T lymphocyte in liver fibrosis model was(7.41±2.15)%,which was significantly lower than that in control group(12.88±2.93)%(t=3.752,P<0.01).Furthermore,the frequency of Tregs in spleen of the model group was(9.49±1.16)%,which was also significantly lower than that in control group(13.16±2.36)%(t=2.793,P<0.05).In addition,the levels of serum ALT,AST and fibrosis parameters were inverselv correlated with the frequency of Tregs in spleens and peripheral blood(ALT and Tregs in blood:r=-0.727,AST and Tregs in blood:r=-0.698,ALT and Tregs in spleen:r=-0.663,AST and Tregs in spleen:r=-0.535,HA and Tregs in blood:r=-0.719,PCⅢ and Tregs in blood:r=-0.558,CⅣ and Tregs in blood:r=-0.792,HA and Tregs in spleen:,r=-0.424,PCⅢ and Tregs in spleen:r=-0.685,CⅣ and Tregs in spleen:r=-0.506;all P<0.05).However,the linear correlations between serum Alb and Tregs in spleens and peripheral bloods were not observed(r=0.423,0.372,respectively,both P>0.05).Conclusion These findings suggest that the reduction of CD4+ CD25+ Tregs probably play an important role in the immunological pathogenesis of liver fibrosis.

Objective To study the protective role of ulinastatin in acute liver failure (ALF) and the effect on the expression of hemeoxygenase-1 (HO-1).Methods Sixty-six S-D rats were divided into three groups:control group,ALF group (model group) and ulinastatin group (intervention group).The rat model of ALF was induced by intraperitoneal injection of D-galactosamine (D-Gal) and lipopolysaccharide (LPS). The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and malondialdehyde (MDA) were detected dynamically after 6,12,24,36 and 48 h injection.HO-1 mRNA expression in liver tissue was determined by reverse transcriptionpolymerase chain reaction (RT-PCR), and the expression of HO-1 protein was detected by immunohistochemistry.The differences among multiple groups were compared by univariate ANOVA and pairwise comparison was done by least significant difference (LSD). Results D-Gal/LPS injections successfully induced ALF rat model which presented with elevated levels of serum ALT,AST and liver MDA after 6 h injections (F=23.864,38.446,18.051,respectively,all P＜0.01),and peaked at 12-24 h after injections.Twenty-four hours after D-Gal/LPS treatment,the levels of serum ALT,AST and MDA in model group and intervention group were (8 346.7±1 363.1) U/L vs(4 151.3±970.0) U/L,(9 766.7±1.274.1) U/L vs (4 696.7±1 476.9) U/L,(8.34±1.13)μmol/g vs (4.66±0.91 ) μmol/g,respectively,which were significantly higher than those e (24.0±2.0) U/L,(82.3±16.9) U/L,(2.55±0.22) μmol/g,respectively] in control group (F=55.684,55.501,47.843,respectively,all P＜0.01);while those in intervention group were much lower than those in model group (P＜0.01).The expressions of HO-1 mRNA and protein in model group were significantly increased than those in control group (P＜0.01),while those in intervention group were even higher (P＜0.01).Conclusion Ulinastain could up-regulate the expressions of HO-1 mRNA and protein,which indicates that ulinastain may play anti-oxidant and anti-inflammatory roles in ALF through HO-1 pathway.

Objective To investigate the relationship of 24 h urinary potassium (K) measurement and the symptoms change in ketamine-associated cystitis.Methods Forty-three ketamine-associated cystitis patients (29 male cases,14 female cases) were analyzed.The average age was 22 (17-29) years.Thirty-two patients without indwelling urinary catheter were categorized as group A,while the other 11 patients with indwelling urinary catheter were in group B.The therapy regimes consisted of anti-inflammatory,antioxidant,relieving spasm and pain,improving the microcirculation and repairing the bladder epithelium barrier.Thirty healthy adults were selected as the controls.Urinary K,sodium (Na) and creatinine (Cr)were determined in 24 h urine samples from all patients and controls before and after treatments.24 h urinary Cr was used as the internal standard.24 h urinary K and Na concentrations of the patients were calculated as relative to the Cr concentrations.The pelvic pain and urgency/frequency symptom (PUF) was used for evaluation before and after the treatments.The differences of urinary K were compared within each group and between groups before and after treatments.In addition,relationship of urinary K and PUF were assessed.Results Urinary Cr concentrations in all groups were not significantly different (P ＞ 0.05).Patients in group A had lower average K-to-Cr ratios than those patients in group B and controls (A 1.80 ± 0.67 vs.B 6.22±0.92 mmolK/mmol Cr,P=0.0001; A 1.80±0.67 vs.controls 6.47 ±0.97 mmol K/mmol Cr,P =0.0001) before treatments.But the ratios of K-to-Cr in group A were not significantly different with group B and controls after treatments (A 6.23 ± 1.42 vs.B 6.02 ± 0.98 mmol K/mmol Cr,A 6.23 ± 1.42vs.controls 6.47 ±0.97 mmol K/mmol Cr,F =0.698,P =0.472).PUF in both groups was not significantly different before treatments.For group A,PUF was negatively correlated with urinary K before and after treatments (before: r=-0.637,P=0.0001; after: r=-0.427,P=0.015).For group B,PUF had no correlation with urinary K before treatment (r=0.581,P =0.188),while there was a negative correlation between them after treatments (r =-0.779,P =0.005).PUF scores in all patients (group A + B)were significantly decreased after treatments when compared to those before treatments (18.12 ± 2.83 vs.22.77 ± 3.63,P =0.0001).Conclusion Urinary potassium measurement may have a role in evaluating the disease status and efficacy of treatments of patients suffered from ketamine-associated cystitis.

OBJECTIVE:To evaluate the clinical effects of urapidil on cardiovascular response induced by sympathetic stimulation during front approach cervical vertebra operation.METHODS:One hundred and twenty patients undergoing front approach cervical vertebra operation were randomly divided into three groups.Patients in URA group treated with0.5mg/kg urapidil and patients in NIC group treated with5?g/kg nicardipine when systolic blood pressure(SBP)and heart rate(HR)increased by sympathetic nerve stimulation during the operation.And patients in COM group only increased their anesthesia degree with isoflurane at the same situation.The patients’SBP and HR were observed and compared among three groups at the time before the sympathetic nerve stimulation,and5minutes and10minutes after the drugs treatment respective?ly.RESULTS:The patients’SBP and HR in URA group had no significantly increase after administration of urapidil.In NIC group,although the patients’SBP had no significantly increase,the patients’HR increased significantly after nicardipine was used.But the patients’SBP and HR increased significantly after improved anesthesia degree with isoflurane.CONCLUSION:Urapidil has better effect on sympathetic cardiovascular response during the front approach cervical vertebra operation com?pared with nicardipine.

OBJECTIVEThe purpose of the present study was to investigate the in vitro effects of baicalin on induction of apoptosis in human prostate cancer cell line DU145.

METHODHuman prostate cancer cell line DU145 was treated with different concentration of baicalin in vitro. The apoptosis rate was determined by FACS analysis, cell cycle distribution was detected by flow cytometry, morphological changes and protein analysis were determined by means of electron microscope techniqueand immunohistochemical techniquerespectively.

RESULT50micromol x L(-1) and 125 micromol x L(-1) of baicalin dose-dependently induced apoptosis and inhibited the proliferation of prostate cancer cell DU145 in a dose and time-dependent manner. DNA flow cytometric analysis indicated that baicalin induced a arrest in G1 phase, showing a typical apoptosis peak. Electron microscopy detected a characteristic appearance of the apoptotic cells morphology. Immunohistochemical analysis revealed that induction of apoptosis by ways of inhibition of the bcl-2, loss of the Bax, and upregulation of Fas.

CONCLUSIONThe results indicate that baicalin may induce apoptosis and inhibit proliferation of prostate cancer cells, and has direct anti-tumor effects on human prostate cancer cells.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Flavonoids ; isolation & purification ; pharmacology ; G1 Phase ; Humans ; Male ; Plants, Medicinal ; chemistry ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Scutellaria ; chemistry ; bcl-2-Associated X Protein ; fas Receptor ; metabolism

A study on the microbial transformation of glycyrrhetinic acid (GA) was conducted by a fungus, Cunninghamella blakesleeana CGMCC 3.970 systematically. After incubation with the cell cultures of C. blakesleeana CGMCC 3.970 at 25 degrees C for 7 days on a rotary shaker operating at 135 r x min(-1), GA was converted into one major product and five minor products. The products were extracted and purified by solvent extraction, macroporous adsorbent resin, silica gel column chromatography, and semi-preparative RP-HPLC chromatography. Their structures were identified as 3-oxo-15α-hydroxy-18β-glycyrrhetinic acid(1), 3-oxo-15β-hydroxy-18β-glycyrrhetinic acid (2), 7β,15α-dihydroxy-18β-glycyrrhetinic acid (3), 3-oxo-7β, 15α-dihydroxy-18β-glycyrrhetinic acid (4), 7β-hydroxy-18β-glycyrrhetinic acid(5) and 15α-hydroxy-18β-glycyrrhetinic acid(6) by the analyses of MS, 1H-NMR and 13C-NMR spectroscopic data respectively. Among them, 2 was a new compound. These results suggest that C. blakesleeana CGMCC 3.970 has the capability of selective ketonization and hydroxylation for GA. [Key words] glycyrrhetinic acid; Cunninghamella blakesleeana CGMCC 3. 970; microbial transformation
Biotransformation ; Cunninghamella ; metabolism ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; metabolism ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization