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.

A strain of Cellulosimicrobium cellulans Ha8 was studied on its morphological, biological characteristics and its utilization of several kinds of benzoic compounds, the results showed this strain was Gram-positive, the long rod-shaped cells were changed into short rod-shape gradually. pH value from pH 6.0 to pH 9.0 and the temperature from 20 ℃ to 40 ℃ were good for its growth. It could not only hydrolyze protein and starch, use cellulose and pectin, decomposite chitin, liquify gelatin and fix nitrogen, but also use phenol, xylene, benzoic, cinnamic acids and diphenlamine as the sole carbon resource for its growth. It could tolerate 0 mmol/L~30 mmol/L, 0 mmol/L~8 mmol/L, 0 mmol/L~30 mmol/L, 0 mmol/L~15 mmol/L and 0 mmol/L ~ 40 mmol/L of benzoic acids, phenol, xylene, cinnamic acids and diphenlamine seperately, but could not use 2,4-dinitrophenol, o-Nitrophenol, 2-Methoxyphenol, aminobenzenesulfonic acid, catechol and o-Phenanthroline as its sole carbon resource.

AIM: To observe the effect of intravenous methylprednisolone combined with peri - orbital injection of triamcinolone acetonide for diffuse - type orbital inflammatory pseudotumor.
METHODS: Diffuse - type orbital inflammatory pseudotumor in 15 cases ( 19 eyes ) were treated. Intravenous implosive methylprednisolone therapy (0. 5g/ d) was used in the first 3d, and 0. 5g once a week in the following 3wk, ended by 0. 25g once a week in the last 6wk, which meant the total dose was 4. 5g and the whole course lasted for 10wk. At the same time, peri - orbital injection of triamcinolone acetonide ( 40mg ) was performed once in every 3wk, totally 2-4 times.
RESULTS: Eight eyes from 7 cases were completely cured, 11 eyes from 8 cases were partly cured. No recurrence and severe complications were observed in the treatment duration.
CONCLUSION: Intravenous methylprednisolone combined with peri - orbital injection of triamcinolone acetonide is effective, safe and feasible in treatment of diffuse type orbital pseudotumor with less complications.

Objective To investigate the effects of ulinastatin on the expression of neutrophil CD_(11b) and CD_(18) during orthotopic liver transplantation(OLT).Methods Forty patients with liver diseases,ASAⅢorⅣ, undergoing OLT were randomly divided into two groups.Ulinastatin group(n=20)received intravenous infusion of ulinastatin 3?10~5 IU in 100 ml normal saline after skin incision and repeated every 4 hours thereafter.Control group received same amount of normal saline instead(n=20).Blood samples were taken immediately before skin incision,120 min after skin incision,30 min of anhepatic phase,60 min of neohepatic phase and at the end of operation for measurement of CD_(11b) and CD_(18) expression of neutrophil by flow cytometry.Results CD_(11b)/CD_(18) expression was increased significantly in control group 60 min of neohepatic phase and at the end of operation compared to the level before skin incision,but in ulinastatin group there was no significant change in CD_(11b)/CD_(18) expression during the whole procedures(P＞0.05).CD_(11b)/CD_(18)expression was significantly lower 60 min of neohepatic phase and at the end of operation in ulinastatin group than in control group(P＜0.01).Conclusion Ulinastatin can inhibit the increase in CD_(11b)/CD_(18)expression during OLT,and be helpful for reducing the inflammatory response.

Objective To explore the effects of peripheral stem cell transplantation on extremely severe bone marrow form of acute radiation sickness.Methods One patient was radiated aecidently with the radiation dosage of 9～15 Gy and diagnosed as extremely severe bone marrow form of acute radiation sickness.Pretreatment was performed at 4th day after the accident and 3 days later,HLA- matched allogeneic stem cell transplantation on the patient was performed.Graft versus host disease (GVHD) was prevented with cyclosporin A (CsA) and mycophenolate mofetil (MMF).Results The haematopoiesis was recovered at 9th day after transplantation.At 11th day after transplantation, WBCs were increased to 14.74?10~9/L and returned to the normal levels subsequently,number of platelets risen to 51?10~9/L and hemaglobin was over 80 g/L.TRS-PCR and blood type dynamic detection testified that the donor's cells were stably implanted,the chromosome aberration and micro- nuclei disappeared after transplantation,and the patient's blood type changed into the donor's at 27th day after transplantation.But the radiation injury were still getting worse complicated with multiple infections,At 68th day after transplantation (75 days after the accident),the patient was died of mul- tiple organs failure.Conclusion Extremely severe bone marrow form of ARS can achieve hematopoie- sis recovery by allogeneic stem cell transplantation,but only hematopoiesis recovery can not cure the immunodefficiency and the radiation injury of the whole body.

BACKGROUND:Our previous study has already manifested that Chinese medicine Xiaokening can effectively prevent and treat the early diabetic nephropathy. OBJECTIVE:To compare the effect of rhein and archen on the apoptosis of mesangial cells of rats cultured with high glucose. METHODS:Mesangial cells were stimulated by different concentrations of rhein and archen (20, 40 and 80 μmol/L). Morphology of apoptotic cells was observed by hematoxylin-eosin staining. Karyon apoptosis was examined by fluorescence staining of DAPI. cellapoptosis rate was detected by flow cytometry. RESULTS AND CONCLUSION:The outcome of hematoxylin-eosin staining and DAPI staining indicated that the effect of rhein was much stronger than that of archen on the apoptosis of mesangial cells in the rats cultured with high glucose. Comparison of the apoptosis rate also indicated that the rhein had a stronger effect on the earlier and the later stage apoptotic rate of mesangial cells than archen. Both rhein and archen with different concentrations can induce apoptosis of mesangial cells, but the effect of rhein is much stronger.

Objective To explore the effects of 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3] on memory CD4+T cells of focal proliferative IgA nephropathy (IgAN)patients.Methods (1) Total of twenty incipient focal proliferative IgAN patients (Lee classification:Ⅲ level) were chosen as IgAN group and 20 healthy volunteers were chosen as healthy control group.The level of serum 1,25(OH)2D3 was measured by radioimmunoassay (RIA).Peripheral blood mononuclear cells (PBMCs) were separated by the method of Ficoll density gradient centrifugation and were stimulated with anti-CD3/anti-CD28 in the absence or presence of various concentrations of 1,25(OH)2D3,Dexamethasone(DEX) and 1,25(OH)2D3and DEX combined.PBMCs were cultured for 72 hours and the levels of IFN-γ,IL-4,IL-17A,Foxp3 were measured by flow cytometry(FCM),standing for the levels of Th1,Th2,Th17,Treg.(2) IgAN group was divided into two subgroups (proteinuria ＜ 1 g/24 h subgroup,proteinuria≥ 1 g/24 h subgroup),then the serum levels of 1,25(OH)2D3,IFN-γ,IL-4,IL-17A,Foxp3 were compared.Results Compared with healthy control group,serum 1,25(OH)2D3 level of IgAN group was significantly lower (P ＜ 0.05).Serum 1,25(OH)2D3 level in proteinuria≥ 1 g/24 h subgroup was significantly lower than proteinuria ＜ 1 g/24 h subgroup and healthy control group (P ＜ 0.05).The level in proteinuria ＜ 1 g/24 h subgroup was lower than healthy control group,but the difference was not statistically significant (P ＞ 0.05).(2) The levels of IFN-γ and IL-17A and the ratios of IFN-γ/IL-4,IL-17A/Foxp3 in IgAN group increased significantly compared with healthy control group (all P ＜ 0.05),and the level of Foxp3 decreased significantly (P ＜ 0.05).The level of IL-4 also increased,but the difference was not statistically significant (P ＞ 0.05).The levels of IFN-γand IL-17A and the ratio of IL-17A/Foxp3 in proteinuria≥ 1 g/24 h subgroup increased significantly,and the level of Foxp3 decreased significantly,compared with urinary protein ＜ 1 g/24 h subgroup and healthy control group (P ＜ 0.05).The ratio of IFN-γ/IL-4 in proteinuria≥1 g/24 h subgroup and proteinuria ＜ 1 g/24 h subgroup all increased,compared with healthy control group,and the ratio in proteinuria≥ 1 g/24 h subgroup increased significantly (P ＜ 0.05).There was no significant difference in the level of IL-4 among all groups.(3) After treatment with 1,25(OH)2D3,the levels of IFN-γ and IL-17A and the ratios of IFN-γ/IL-4 and IL-17A/Foxp3 decreased significantly,and the level of Foxp3 increased significantly (P ＜ 0.05),and these effects were more obvious as the increase of the drug concentration.The level of IL-4 did not change significantly.The combination of 1,25(OH)2D3 and DEX had a synergistic inhibition on the production of IFN-γ,IL-4,IL-17A,and the ratios of IFN-γ/IL-4 and IL-17A/Foxp3,and had a synergistic promotion on the production of Foxp3.Conclusions There is a certain extent of vitamin D deficiency in focal proliferative IgAN patients,which may be associated with the severity of proteinuria.The disorder of immunomodulatory effects of memory CD4+ T cell might exist in the patients of focal proliferative IgAN.1α,25-dihydroxyvitamin D3 might have beneficial effects on the immunoregulation of memory CD4+T cells of focal proliferative IgAN patients.