OBJECTIVETo investigate the apoptosis regulation on hepatoma cells by HBx between genotype B and C.

METHODSGenotype B and C HBx gene fragments were amplified and inserted into green fluorescent protein (GFP) eukaryotic expression vector pEGFP-C1 to construct recombinant pGFP-XB and pGFP-XC. The pEGFP-C1, pGFP-XB and pGFP-XC were introduced into Bel-7402 cells by Fugene HD to obtain Bel-7402 cells expressing GFP. The transcription and expression of HBx gene were demonstrated by RT-PCR and Western Blot analysis. Bel-7402, Bel-7402/GFP, Bel-7402/GFP-XB, Bel-7402/GFP-XC cells were treated with adriamycin (2.5 microg/ml), and the apoptosis of the cells was determined by trypan blue exclusion, and flow cytometry analysis.

RESULTSRT-PCR and Western Blot analysis showed that HBx genes of genotypes B and C were transcribed and expressed in Bel-7402/GFP-XB, Bel-7402/GFP-XC cells. Trypan blue exclusion showed adriamycin induced time-dependent cell death in Bel-7402, Bel-7402/GFP cells while no significant cell death was observed in Bel-7402/GFP-XB, Bel-7402/GFP-XC cells. Flow cytometry analysis indicated that no significant differences of apoptosis rates of Bel-7402/GFP-XB (3.87%) and of Bel7402/GFP-XC (4.01%) were observed (P > 0.05), moreover, no significant differences of Bel-7402/ GFP-XB (3.87%), Be17402/GFP-XC (4.01%) and of the untreated cells. Apoptosis rates in Bel-7402/GFP-XB (3.87%), Bel-7402/GFP-XC (4.01%) cells were significantly lower than those in Bel-7402 (27.05%) and Bel-7402/GFP (29.14%) cells at 48 hours after the adriamycin treatment (P < 0.01).

CONCLUSIONSBel-7402 cell lines expressing GFP, GFP-XB and GFP-XC fusion proteins were successfully established. HBV X protein blocks adriamycin-induced apoptosis of Bel-7402 cells. There is no difference between HBx of genotype B and C in inhibiting apoptosis induced by adriamycin.

Antiviral Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Hepatitis B ; drug therapy ; physiopathology ; virology ; Hepatitis B virus ; classification ; drug effects ; genetics ; metabolism ; Humans ; Trans-Activators ; genetics ; metabolism

Objective To observe the influences of electroacupuncture(EA) on the morphological changes in neurons and glia cells in sacral dorsal commissural nucleus(SDCN) of rats with ulcerative colitis(UC),and study the mechanism of acupuncture in modulating central nervous system in UC.Methods Forty-eight male SD rats were randomly assigned to four groups: control group,UC model group,EA Tsusanli group(UC plus EA at Tsusanli acupoint) and EA non-acupoint group(UC plus EA non-acupoint group).According to the acupuncture duration,EA Tsusanli group was divided into three subgroups: 1-day,3-day and 5-day subgroup(8 rats for each subgroup).UC model of rats was reproduced by instillation of a solution containing ethanol and 2,4,6,-trinitrobenzenesulfonic acid(TNBS) into the distal colon.When the treatment was ended,all the rats were sacrificed and SDCN slides were prepared.The expressions of Fos,glial fibrillary acidic protein(GFAP) and OX42 immunoreactivity in SDCN were assessed and calculated by using single or double immunohistochemical methods.Results In the three experimental groups,the Fos-positive neurons,GFAP-positive astrocytes and OX42-positive microglia were mainly expressed in SDCN.In UC model group and EA non-acupoint group,the expressions of Fos,GFAP and OX42 immunoreactivity were stronger than those in EA Tsusanli group(P

Objective To evaluate the apical sealing ability of iRoot SP root canal sealer in oval-root canal. Methods After root canals were instrumented by Mtwo, 28 premolars with oval-root canal were randomly divided into 2 groups, 14 teeth in each. The root canals were obturated with AH Plus (group A) and iRoot SP (group B) by modified continuous wave condensation technique. The apical leakage was evaluated by dye penetration method and transparent teeth technique. Results Mean depth of leakage was (0.92 ± 0.26) mm for group A and (0.84 ± 0.40) mm for group B. There was no significant difference between the two groups (t=0.506, P > 0.05). Conclusion The apical sealing ability of iRoot SP is equivalent to AH Plus in oval-root canal.

Objective To detect the expression of E-cadherin and ?-catenin in breast carcinoma and analyze the relationship between E-cadherin and ?-catenin and the clinicopathological features. Methods The expression of E-cadherin and ?-catenin in breast cancer, paracarcinoma breast tissue, simple mastoplasia and atypical mastoplasia were detected by immunohistochemical method and the results were compared. Results The abnormal expression rates of E-cadherin and ?-catenin in breast cancer tissue were 51.9% and 61.1 %,respectively. Abnormal expression of E-cadherin was significantly correlated with histological grade. Abnormal expression of ?-catenin was significantly correlated with TNM staging, axillary lymph nodes metastasis and postoperative distant metastasis. COX multiple factor analysis showed that neither E-cadherin nor ?-catenin expression was an independent indicator for the prognosis of breast cancer. Conclusions Abnormal expressions of E-cadherin and ?-catenin are correlated with occurrence and development of breast carcinoma. Abnormal expressions of E-cadherin and ?-catenin are good indicators to judge invasion and metastasis of breast carcinoma.

OBJECTIVE: To provide references for the evolvement of the administration of drug labels and package inserts in China. METHODS: By content analysis in non-contact research, 23 law documents relevant to American drug labels and package inserts were collected from FDA website, of which, 7 were acts, 1 federal regulation and 15 guiding principles RESULTS: The drug labels and package inserts of the prescription drugs and the over-the-counter drugs in US were administrated in classification with an emphasis on the contents and formats. CONCLUSIONS: A set of comprehensive lawful system for drug labels and package inserts has been established in American, from which China should draw experience to ensure the comprehensiveness, objectiveness and accuracy of drug information and guarantee safety and efficacy of drugs.

Cochlea ; metabolism ; Deafness ; genetics ; Humans ; Mutation

Objective To investigate the effects of electro-acupuncture on gastric emptying and Fos expression in the neuron of vagal-solitary complex (VSC) of rat bulbus after lipopolysaccharide (LPS) intraperitoneal injection (i.p.). Methods 40 male SD rats were randomly assigned to 4 groups: control group, LPS i.p. group, LPS i.p. plus electro-acupuncture at Tsusanli point group and LPS i.p. plus electro-acupuncture at non-meridian-non-acupoint group, 10 rats for each group. Immunohistochemical techniques were used to detect the Fos expression in VSC. The animal's gastric emptying was measured by phenol red method. Results The rats with gastric emptying decreased greatly to (20.7±4.5)% 2.5 hours after LPS injection, and Fos-positive neurons were significantly found in VSC (83.2±6.6) compared with control group. However, in the group of LPS i.p. plus electro-acupuncture at Tsusanli point, the gastric emptying was up-regulated to (44.1±6.2)%, and the expression of Fos -positive neurons were down-regulated to (37.9±3.8) compared with LPS i.p. group. No significant difference was found between the group of LPS i.p. plus electro-acupuncture at non-meridian-non-acupoint and the group of LPS i.p. Conclusion LPS i.p. can retard the gastric motility in rats, electro-acupuncture at Tsusanli point may well regulate the function in LPS model rats. This function may be connected with its protective effects on Fos immunoreactive neurons activity in VSC of rat bulbus.

BACKGROUND:Currently, one of common methods is discectomy, nerve root decompression and fusion rigid fixation from the midline approach for disc herniation which is inefficient by conservative treatments. Thus, it is causing degeneration and limiting lumbar physiological activity of adjacent segments. The treatment of non-fusion lumbar disc herniation with the traditional posterior midline incision approach has some disadvantages such as big incision, wide peeling, and back muscle denervation.
OBJECTIVE:To observe therapeutic effects of dynamic stabilization system through Wiltse approach on lumbar disc herniation, and to compare the outcomes with traditional posterior approach.
METHODA total of 46 patients, who had undergone discectomy and internal fixation using dynamic stabilization systems for lumbar disc herniation at the Renmin Hospital of Wuhan University from January 2011 to January 2013, were enrol ed in this study. The operation was performed through the traditional posterior approach in 25 patients and Wiltse approach in 21 patients.
RESULTS AND CONCLUSION:Al 46 patients were fol owed up for 7 to 31 months (averagely, 13.8±2.4 months). The length of incision, intraoperative blood loss, and postoperative drainage amount were less in the Wiltse approach group than in the traditional posterior approach group (P<0.05). No significant difference in visual analog scale scores and operative time was detected between the two groups at 1 week and 6 months after fixation (P>0.05). Radiographs revealed that the position of implants was good in al patients, no loosing or breakage. These data verified that the early effect of dynamic stabilization system through Wiltse approach for lumbar disc herniation is similar to that of traditional posterior approach.

AIM: To study the processing of Indigo Naturals. METHODS: Study on the microstructure of Indigo Naturals with scanning electron microscope (SEM) and X ray diffractometer (XRD) were carried out. RESULTS: Through the SEM, it is clear that Indigo attached to the surface of some white substance, and the XRD proves that the white substance in Indigo Naturals is calcium carbonate (CaCO 3). CONCLUSION: The study about the processing of Indigo Naturals indicates that Indigo Naturals consists of Indigo and other effective compositions attached the surface of calcium carbonate (CaCO 3).

Objective To study the effect of total nutrient admixture (TNA) on lipid metabolism in rats with acute hepatic failure (AHF). Methods Fifty-six male Wistar rats were randomly divided into four groups: normal diet group, nitrogen-free diet group, fat-free nutrient admixture group, and TNA group. All rats were injected with D-galac- tosamine to induce AHF model Meanwhile, 6 healthy Wistar rats were used as control group. TNA provided energy 221.75 kJ (1 kcal=4.184 kJ), nitrogen 0.365 g, nitrogen to energy ratio 1:145, amino acids 2.28 g (including branched-chain amino acid 0.70 g), glucose 7.85 g, and fat 1.25 g on a daily basis. After 10 days of the operation of Jugular vein puncture tube, blood glucose, blood lipid, liver and renal function were determined. Results The blood glucose level was significantly higher in TNA group than that in fat-free nutrient admixture group (P < 0.05). The levels of serum triglyceride and cholesterol were highest in fat-free nutrition admixture group, and triglyceride level was signif- icantly higher than that in normal diet group and nitrogen-free diet group (P <0.05). The levels of serum triglyceride and cholesterol were significantly higher in TNA group than those in normal diet group (P <0.05). The levels of ala- nine aminotransferase, alkaline phosphatase, total bilirubin and blood urea nitrogen were lower in TNA group than those in fat-free nutrient admixture group (P < 0.05). Conclusion The nutritional proportion of TNA is suitable for the metabolism disorder of hepatic failure, and therefore TNA can reasonably promote the anabolism and reverse the deteri- oration of hepatic failure in rats.