Objective To investigate the dynamic changes of metallothionein(MTs)gene expression and explore the important significance of MTs during hepatocarcinogenesis.Methods One hundred and twenty-five SPF 5 -8-week old male C57BL/6J mice were randomly divided into control group and model group.Diethylnitrosamine (DEN) was given to the mice at a dose of 100 mg/kg, ip, and 50 mg/kg, ip, in the first and next week, respectively.The mice were given ethanol (53%, 5 mL/kg/day, 5 days/week) from the third week of experiment till 35 weeks.At 1, 3, 9, 13, 24 and 35 weeks of the experiment, liver samples were taken for histopathological examination of liver damages and incidence of HCC. The liver index and malondialdehyde (MDA) of liver homogenate were determined.All liver tissue samples were examined by histopathology using hematoxylin and eosin (HE), Masson and reticular fiber staining.Real-time RT-PCR was used to analyze the mRNA expression level of liver metallothionein-1 /2 (MT-1 /2) in different periods.Results Progressive liver damages in model group mice were identified in different periods.Hepatocytes abnormal tission and abnormal liver plate structure, architecture often characteristic of HCC could be seen in approximately 50% of mice at 35 weeks.In addition to these, a higher liver index also were seen at 35 weeks.Increased MDA levels in the mouse liver tissues were observed in each stage.Real-time RT-PCR analysis showed that significantly increased transcription of MT-1 and MT-2 at 1, 3 and 9 weeks, then gradually declined and even below the normal level.Conclusions MTs gene expression levels in mouse liver tissues are changed from significantly increased in the early stage of injury to decreased expression combined with distinct fibrosis. Our findings further demonstrate that the down-regulation of MTs level is closely correlated with hepatocarcinogenesis.

[Objective]To investigate the differentiation of acute from chronic osteoporotic vertebral fractures.[Method]The clinical presentation,X-ray,CT and MRI of patients with osteoporotic vertebral fractures were evaluated in the study.[Result]In 36 cases,64% were identified as acute and 36% with chronic osteoporotic vertebral fractures.The decreased anterior vertebral height was a most important criterium for the diagnosis of osteoporotic vertebral fractures.Degenerative changes on the X-ray film were usually found in chronic fractures.A high intense signal in a specific T2 under eliminating fat tissue was always observed in acute fracture.[Conclusion]The local back pain located at the level of the fractured vertebral body suggests an acute vertebral fracture.Degenerative changes occurred in the edged vertebra is indicative of a chronic fracture.The finding of a specific T2 high intense signal under eliminating fat tissue on MRI is a gold standard for the diagnosis of an acute vertebral fracture.

Purpose To evaluate the application of mismatch repair (MMR) genes proteins expression and methylationspecific to screen for Lynch syndrome patients.Methods 126 endometrial carcinoma patients were tested the protein expression of hMSH2,hMSH6h,hMLH1,hPMS2 by immunohistochemically of SP,and the methylation status of hMLH1 genes by the methylation-specific PCR.Results The result of MMR immunocytochemistry showed that 22％ (28/126) cases lacked MMR protein expression,including hMLH1-/hPMS2-in 12 cases,4 hMSH2-/hMSH6-,6 hPMS2-,3 hMLH6-and 3 hMLH1-.Meanwhile,the methylation-specific PCR test showed that 9 cases was methylation status of hMLH1 genes in 15 cases hMLH1-,and suggested the patients might be sporadic endometrial carcinoma.Conclusion Immunohistochemical of SP staining for MMR proteins in endometrial carcinoma patients,accompanied by testing for the methylation status of hMLH1 genes,may be an effective approach to screen for Lynch syndrome.

Objective To examine the effects of different doses of midazelam on ED50 of emulsified isoflurane and to determine the type of interaction between them for hypnosis by isobelographic analysis in rats. Methods One hundred and twenty-five adult male SD rats weighing 240-300 g were randomized into 5 groups (n=25 each): group Ⅰmidazolam (M); group Ⅱ emulsified isnflurane (Ⅰ); group Ⅲ ,Ⅳ ,Ⅴ, 1/4, 1/2 and 3/4 ED50 of midazelam for hypnosis + emulsified isoflurane (MI1 , MI2, MI3). Up-and-down sequential experiment was used to determine ED50of midazolam and emulsified isoflurane for loss of righting reflex in group Ⅰ -Ⅴ . The intial dose of midazolm was 17.3 mg/kg in group M. The initial doses of emulsified isoflurane (120 mg/ml) were 0.55 (in group Ⅰ, 0.22 (MI1), 0.19 (MI2) and 0.12 ml/kg (MI3) respectively. In group MI1-3 midazolam was injected over 15 seconds and after an interval of 2.5 min emulsified isoflurane was injected over 10 s. ED50 was calculated using Dixon-Mood method. Isebolographic and algebraic analyses were used to determine the type of interaction between midazolam and emulsified isoflurane. Results The five groups were comparable with respect to M/F sex ratio and body weight. The Edso of midazolam was 26.0 mg/kg in group M. Midazolam 6.5, 13.0 and 19.5 mg/kg were given in group MI1 , MI2 and MI3 respectively. The ED50 of emulsified isoflurane was 0.67 (ingroup Ⅰ), 0.30 (MI1), 0.22 (MI2) and 0.18 ml/kg (MI3) respectively. The isobolographic analysis indicated that with increasing doses of midazolam, the Edw of emulsified isoflurane decreased progressively in a non-linear fashion. The isobolographic and algebraic analyses demonstrated that the interaction between midazolam and emulsified isoflurane was synergistic for hypnosis. Conclusion The hypnosis is synergistic when midazolam 6.5,13 mg/kg are combined with emulsified isoflurane and additive when midazolam 19.5 mg/kg is combined with emulsified isoflurane.

OBJECTIVE:To compare home-made mu'an-eye-gel(acyclovir plus honey) with commercial aciclovir(ACV)-eye-gel in releasing drug characteristics in vitro.METHODS:The in vitro drug release test was conducted by the third method of dissolution determination stated in Chinese Pharmacopeia together with bag filler method.The cumulative drug-releasing percentage and the acyclovir amount in mu'an-eye-gel versus ACV-eye-gel were determined by UV spec-trophotometry,and the accumulative releasing drug percentages of the two preparations were computed and their drug release behaviors w ere compared.RESULTS:The in vitro releasing behaviors of mu'an-eye-gel followed the Weibull kinetic equa-tion,however the vitro releasing behavior of commercial ACV-eye-gel followed the zero order kinetic equation,and the T80%and Q8 h had statistical significances between(mu'an-eye-gel:T80%=3.156?0.013(h),Q8 h=93.28?0.010(%);ACV-eye-gel:T80%=10.16?0.009(h),Q8 h=67.85?0.025(%)) 2 kinds of preparation.CONCLUSION:Mu'an-eye-gel is superior to the commercial ACV ophthalmic gel in both releasing velocity and accumulative drug release percentage.

OBJECTIVE:To investigate whether the magnesium sulfate injection(MSI)into cisterna magna in a rabbit with subarachnoid hemorrhage(SAH)can reverse the cerebral vasospasm and damage of brain tissues.METHODS:The single-hemorrhage SAH rabbit model was used.Thirty New Zealand white rabbits were randomly divided into the following three groups(n=10,in the each group):sham group,model group and MSI group.At 24 hours after SAH,the rabbits were injected with 0.1 mL?kg-1 of saline into cisterna magna in sham and model groups,while 0.1 mL?kg-1 of 4% magnesium sulfate was injected into cisterna magna of rabbits in MSI group.All animals were sacrificed at 48 hours after SAH.Basilar arteries and hippocampus were then removed for measurement of the cross-sectional areas of basilar arteries and the hippocampus normal neuron density of CA1 regions.RESULTS:The cross-sectional areas of basilar artery and the hippocampus normal neuron density of CA1 regions were smaller in model group than in the other two groups(P0.05).CONCLUSIONS:This study results indicate that injecting magnesium sulfate into cisterna magna can reverse the cerebral vasospasm and the following hippocampal neuron damage in rabbits with subarachnoid hemorrhage.

Objective To investigate the effect of triptolide on the proliferation and apoptosis of human epidermal squamous cell carcinoma cell line A431 in vitro. Methods Human epidermal squamous cell carcinoma cell line A431 was cultured. After the treatment with triptolide, the inhibi-tion of cellular growth was determined by measuring MTT dye absorption of the living cells. Light mi-croscope showed morphological changes. The cell cycle and apoptosis rate were assessed by flow cy-tometry. Results Triptolide could significantly inhibit the proliferation of A431 cells in a dose- and time-dependent manner. Triptolide could also cause cell morphological changes ( the number of float-ing cells and nuclear pyknosis increase), induce cell apoptosis, and change the distribution of cell cycle phase in A431 cells. Compared with the control group, the G0/G1 phase A431 cell rate in-creased and the rate of S phase cell decreased in TP-treated group. Cell cycles were obviously inhibi-ted by triptolide in G0/G1 phase (both P<0.05). Conclusion TP could play an anti-tumor role by effectively inducing cell apoptosis and inhibiting the proliferation of A431 cells.

Objective To study the activation of Janus protein tyrosine kinase (JAK)/signal transducer and activator of transcription 1 (STAT1) signaling pathway and its inhibitor-signal transducer and activator of transcription-1(SOCS-1) in patients with systemic lupus erythematosus. Methods A total of 45 patients with active systemic lupus erythematosus (SLE) and 30 healthy controls were randomly selected. Western blot was performed to measure the expression of Stat1 protein and phospho-Stat1 protein (an activated form of Stat1 protein) in the monocytes after stimulation with recombinant high mobility group box1 (rHMGB1) at various time points. Expression of Stat1 protein in the skin or lesional skin was also detected. Phasic expressions of SOCS-1 mRNA in the monocytes after rHMGB1 stimulation were detected by real-time reverse transcription-polymerase chain reaction. SOCS-1 gene expression in the skin or lesional skin was also detected. Results The expression level of Stat1 proteins in the monocytes from patients with SLE was higher than that from healthy controls (t=9.16,P＜0.01) and positively correlated with SLE disease activity index (SLEDAI) (r=0.59,P＜0.01). Expression of phospho-Stat1 in the monocytes from SLE patients was time-dependently upregulated after stimulation with rHMGB1 at various time points, while expression of SOCS-1 mRNA remained unchanged(all P＞0.05). Expressions of phospho-Stat1 protein and SOCS-1 mRNA in the monocytes from healthy controls were increased transiently after stimulation with rHMGB1(all P＜0.05). Both expressions of phospho-Stat1 protein and SOCS-1 gene in the lesional skin from patients with SLE were upregulated compared with those in normal skin from healthy controls (all P＜0.01). Conclusion There are hyperactivation of JAK-STAT1 signaling pathway and negative feedback down-regulation of SOCS-1 in patients with systemic lupus erythematosus. HMGB-1 may be partly involved in the pathogenesis of SLE by the abnormal mediating function of JAK-STAT1 signal transduction pathway.

Objective To investigate the influence of CO2-insufflation pressure on invasion potential of the colon cancer cells.Methods With an in vitro artificial pneumoperitoneum model,SW1116 human colon cancer cells were exposed to CO2-insufflation of 5 different pressure groups:6,9,12,15 mm Hg and control group,respectively for 1 h.The invasion capacities of SW1116 cells exposed to CO2-insufflation of 5 different pressure groups were detected by cell adhesion/invasion assay in vitro.Results Immediately following exposure to 15 mm Hg CO2 insufflation,the invasion of SW1116 cells decreased significantly compared to the cells before exposure.At the 0 h time point,the cells exposed to 15 mm Hg were significantly less invasive than those exposed to the other insufflation pressure(P