AIM: To establish the technique of precision-cut fibrotic liver slice (PCLS) and grope the optimal cultural conditions for researching the liver xenobiotic metabolism in vitro and the drug interaction. METHODS: Complex factors (higher fat diet, alcohol and CCl 4) were used to make the animal model of liver fibrosis. Fibrotic liver slices were prepared and cultivation system was established. Lactate dehydrogenase (LDH) leakage, glutathione S-transferase (GST) activity and 3[4,5-Dimethythiazole-2-yl]-2,5- diphenyltetrazolium bromide (MTT) reduction were chosen as indexes to assess the viability of the slice in different thickness, medium pH and cultural time. RESULTS: Rats were in earlier hepatic fibrosis after administration for 3 weeks. When the thickness of slices was 300 ?m and medium pH was 7.0, the LDH leakage, GST activity and MTT reduction could maintain on a steady level in 6 h. CONCLUSION: A 300 ?m of thickness, 7.0 of medium pH and 6 h of cultural time are the optimal slicing and culturing conditions for fibrotic liver slice.

Analysis of metabolic activities of cytochrome P450 isoenzyme is a crucial index to study drug/toxicant metabolism, drug-drug interaction, polymorphism and et al. Due to this practice, it is important to use the proper probe substrate and to conduct the experiment under optimal conditions. The validation information in literatures on the most common and newest in vitro probe substrates have been reviewed.

Objective To observe the clinical effects of short-term intermittent ganeiclovir treatment on infants with cytomegalovirus (CMV) hepatitis. Methods The clinical data of infants with CMV hepatitis were analyzed retrospectively. The liver functions including total bilirubin (TBil), direct bilirubin (DBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and γ-glutamyltransferase (γ-GT) of the patients in treatment group (85 cases) and control group (37 cases) were collected before and after treatment. Meanwhile, the side effects of ganciclovir during treatment were observed. The measurement data were compared by analysis of variance and numeration data were compared by chi-square test. Results After short-term intermittent ganciclovir treatment in treatment group, TBil level was decreased from (109.1±677.8)μmol/L to (62.9±68.1)μmol/L (F=15.34,P<0.01); ALT level was decreased from (160.2±395.3) U/L to (68.1±56.0) U/L (F=4.73, P<0.05). In control group, TBil level was decreased from (94.9±647.4)μmol/L to (49.2±631.5) μmol/L (F=14.80, P<0.01) ; while ALT level was decreased from (131.6±206.2) U/L to (55.3±631.2) U/L (F=3.50, P=0.067). The readmission rate in control group was significantly higher than that in treatment group (21.6% vs 10.6%). Only one case (0.8%) received three times of intermittent ganciclovir treatment. The longest hospitalization time was six weeks. Conclusions Short-term intermittent ganciclovir treatment may be more suitable for infants with CMV hepatitis. There is no obvious side effect observed during the treatment and the hospitalization time can be shortened.

Objective:To investigate the clinical value of the changes of serum complement,immunoglobulin and inflammatory cytokines in children with mycoplasma pneumonia (MP).Methods:60 cases of children with MP infection were collected as MPP group,60 cases of healthy children as control group.The serum levels of complement (C3,C4) and immunoglobulin (IgA,IgG,IgM) were detected by INA,the serum levels of inflammatory cytokines (IL-8,IL-10,IL-13,TNF-α) were detected by ELISA.Results:①The serum levels of IgM,C3,C4,IL-8,TNF-et and IL-13 in MPP group in acute stage were significantly higher than those in the recovery stage and control group (P＜0.05),the levels of IL-8,TNF-et,IL-13 in the recovery stage were significantly higher than those in control group (P＜0.05);the serum levels of IgA,IL-10 in MPP group in acute stage were significantly lower than those in the recovery stage and control group,and these were still significantly lower than the control group (P＜0.05);the IgG in MPP group in acute stage was not significant difference with control group (P＞0.05),but it in recovery stage was significantly higher than the acute stage and the control group (P＜0.05).②The serum levels of IgM,IgG,C3,C4,IL-8,TNF-et and IL-I 3 in MPP group in severe were significantly higher than those in the mild stage and control group (P＜0.05),and the levels of IgM,IL-8,TNF-αt,IL-13 in the severe stage were significantly higher than those in control group (P＜0.05);the serum levels of IgA,IL-10 in MPP group in severe stage were significantly lower than those in the mild stage and control group,and these in mild stage were still significantly lower than the control group (P＜ 0.05).Conclusion:Children with MP infection have a significant cellular and humoral immune dysfunction,the detection of the changes of serum complement,immunoglobulin and inflammatory cytokines is valuable to assessing the disease staging and degree.

Objective To assess the effect of peroxisome proliferator-activated receptor γ (PPARγ) agonist on prostate epithelial cells in vitro.Methods The expression of peroxisome proliferator-activated receptor γ(PPARγ) was studied by immunocytochemistry and immunofluorescence study.The RWPE-1 human prostate epithelial cell line was treated with PPARγ agonist rosiglitazone 100 μmol/L for 48 h.Analysis of apoptosis was performed by Caspase 3/7 activity assay.Mitochondria depolarization was measured by using the potential-sensitive color,JC-1.The expression of apoptosis-related proteins-Bax was investigated by immunohistochemistry.Results PPARγ mainly located in nucleus and perinucleus.RWPE-1 cell line treated with PPARγ agonist rosiglitazone showed higher Caspase 3/7 activity (10636±1032 RLU) than in control (5936±620 RLU),P＜0.01 and significantly upregulated Bax level (8250±694 vs.6017±563)than in control group,P＜0.01.In addition,mitochondrial membrane potential was depolarized in rosiglitazone treated cells.Conclusions PPARmay play important roles in the pathophysiology of BPH.The mechanism might be that PPARγ regulates cell apoptosis.It is suggested that the mitochondrial and Bax pathway might be involved in signaling PPARγ induced cell apoptosis.

Objective To investigate the role of CD4+CD25+FoxP3+ in severe Mycoplasma pneumonia among children.Methods One hundred and forty children with M.pneumoniae pneumonia (65 severe and 75 non-severe) who were hospitalized were enrolled along with forty other children as controls.X-ray was assessed.The proportions of peripheral blood CD4+CD25+FoxP3+cells were determined by flow cytometry.Results Both severe and non-severe children had decreased CD4+CD25+FoxP3+cells as compared with control subjects in acute phase (0.87 ± 0.66％ vs.3.88 ± 2.00％,P ＜ 0.01 and 1.17 ± 0.70％ vs.3.88 ±2.00％,P ＜0.01,respectively).The levels of CD4+CD25+FoxP3+cells in severe children were lower than those in non-severe children in acute phase and recovery phase (0.87 ±0.66％ vs.1.17 ±0.70％,P ＜0.05 and 1.66 ±0.85％ vs.3.61 ± 1.45％,P＜0.01,respectively).Both severe children and non-severe children expressed higher CD4+CD25+FoxP3+cells in recovery phase than in acute phase (1.66 ± 0.85 ％ vs.0.87 ± 0.66％,P ＜0.01 and 3.61 ± 1.45％ vs.1.17 ±0.70％,P ＜0.01,respectively).Conclusion The expression of CD4+CD25+FoxP3+Tregs may play a role in the onset of severity of mycoplasma pneumonia and the low express of CD4+CD25+FoxP3+Tregs in children infected with M.pneumonia may increase the susceptibility to severe mycoplasma pneumonia.

Objective T o evaluate the application value for the prognosis of m echanical ocular injury cases using ocular traum a score (O TS). Methods Four hundred and eleven cases of m echanical ocular traum a w ere retrospectively review ed. O f the 449 eyes, there w ere 317 closed globe injury and 132 open globe injury. O T S variables included num erical values as initial visual acuity, rupture, endophthalm itis, perforat-ing or penetrating injury, retinal detachm ent and relative afferent pupillary block. T he differences be-tw een the distribution of the final visual acuity and the probability of standard final visual acuity w ere com pared to analyze the correlation betw een O T S category and final visual acuity. T he different types of ocular traum a w ere com pared. Results C om pared w ith the distribution of final visual acuity in standard O T S score, the ratio in O T S-3 category w as statistically different in present study, and no differences w ere found in other categories. Final visual acuity show ed a great linear correlation w ith O T S category (r=0.71) and total score (r=0.73). C om pared w ith closed globe injury, open globe injury w as generally associated w ith low er total score and poorer prognosis. R upture injury had poorer prognosis com pared w ith penetrating injury. Conclusion T he use of O T S for the patients w ith ocular traum a can provide re-liable inform ation for the evaluation of prognosis in forensic m edicine.

Objective To investigate the relationship between retinal nerve fiber layer thickness and peripheral neuropathy in elderly patients with type 2 diabetes.Methods Clinical data of elderly patients with T2DM in Endocrinology Department in Beijing Hospital were retrospectively collected.Global and sectorial retinal nerve fiber layer(RNFL)thicknesses were measured by using optical coherence tomography(OCT),and never conduction velocity measurements were performed in all patients.Diabetic peripheral neuropathy was diagnosed by the criteria in diabetic neuropathies update(2010,American Diabetes Association).RNFL thickness was compared between diabetic nonperipheral neuropathy group (control group,n =30)and diabetic peripheral neuropathy group (DPN group,n=17)and between different sub-groups.Results The RNFL thicknesses of temporal,nasal,superior and inferior visual fields and the mean RNFL thickness were less in DPN group than in control group,among which there were significant differences in the RNFL thicknesses of superior,inferior visual fields and the mean RNFL thickness [(107.7±27.4)μm vs.(128.1±17.3)μm,(112.9 ±20.8)μm vs.(130.8±21.8)μm,(88.2±15.5)μm vs.(100.5± 11.3)μm,F=7.446,7.468,7.988,respectively,P=0.009,0.009 and 0.007].RNFL thickness was decreased along with the aggravation of DPN from the control group,the subclinical DPN group to the DPN group successively(all P＜ 0.05).Conclusions Retinal nerve fiber layer thickness is associated with diabetic peripheral neuropathy in elderly patients with type 2 diabetes,and the relationship is more significant in patients with serious DPN.

Objective To explore the protective effect of L-carnitine on neonates with myocardial injury caused by as?phyxia. Methods Forty-four neonates with myocardial injury caused by asphyxia were randomly divided into L-carnitine treatment group (21 cases) and control group (23 cases). Patients in control group were received routine treatment and pa?tients in treatment group were given L-carnitine 0. 1 g/(kg · d) on the basis of routine treatment for 7 days. Symptoms and physical signs were observed before therapy and during the treatment in two groups. Before and after the treatment, plasma levels of free L-carnitine and cardiac troponin I (cTnI) were detected with the method of colorimetric assay and chemilumi?nescent, respectively. Results The clinical effective rate was significantly higher in treatment group than that of control group (90.48%vs 60.87%, P<0. 05). Compared with the control group, there was a significantly higher plasma concentra?tion of free L-carnitine in treatment group after treatment [(27.00±5.69)μmol/L vs (13.20±3.04)μmol/L, P<0.05]. In treat?ment group, plasma concentration of free L-carnitine was significantly higher after treatment than that of pre-therapy [(14.87 ± 3.95)μmol/L,P<0.05]. Compared with the control group, there was a significantly lower plasma concentration of cTnI after treatment in treatment group [(0.025±0.006)μg/L vs (0.046±0.010)μg/L, P<0.05]. In the treatment group, there was a significant correlation between decreased plasma concentration of cTnI and increased plasma concentration of free L-carnitine (r=0.899, P<0.05). Conclusion Administration of L-carnitine can effectively decrease the abnormal plasma lev?el of cTnI in neonates with myocardial injury caused by asphyxia, and thereby protect the myocardium.

Objective To construct expression vectors of human ribosomal protein S 3(RPS3) and RPS3Ser209 mutant in orcler to investigate the effect of RPS3Ser209 mutant on NF-κB signaling pathway and DNA binding capacity .Methods The vector RPS3-myc was amplified by polymerase chain reaction ( PCR) from the human liver cDNA and subcloned into pcDNA-3.1myc-HisB.RPS3S209A represented mutant RPS3 expression vectors, in which the designated amino acid was mutated to an alanine residue .Dual luciferase reporter gene assay was used to detect the NF-κB transcription activity in HEK293 cells,immunofluorescence to detect RPS3 location, and EMSA to examine NF-κB DNA-binding activity.Results The expression vectors of RPS3-myc and RPS3S209A-myc were constructed.Compared with wild-type RPS3,the nucleus translocation, transactivation activity of NF-κB and DNA binding ability of RPS3S290A were reduced significantly .Conclu-sion The impact of RPS3 on NF-κB signaling pathway depend on its serine 209.