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

Mammal multidrug and toxin extrusion proteins (MATEs) play an important role in the transport of organic cations in the body. MATEs mediate the final excretion step for multiple organic cation drug used clinically and important endogenous substances. This article reviews the discovery, type, gene coding and polymorphism, body distribution, classification of substrates and inhibitors and their research method of MATEs. The article also discusses the major research significance of MATEs with examples.
Animals ; Biological Transport ; Cations ; Organic Cation Transport Proteins ; metabolism ; Polymorphism, Genetic

Abscess ; diagnosis ; microbiology ; surgery ; Bacteria, Aerobic ; isolation & purification ; Child, Preschool ; Female ; Fever ; Humans ; Prognosis ; Spleen ; pathology ; surgery ; Splenic Diseases ; diagnosis ; surgery ; Treatment Outcome

Objective To evaluate the relationship between recurrence of cryptogenic ischemic cerebrovascular disease (CICVD) and patent foramen ovale (PFO),as well as to access the clinical significance of PFO in ischemic cerebrovascular disease.Methods Consecutive patients with CICVD aged 15 to 70 years who were hospitalized in Department of Neurology,Xuanwu Hospital Capital Medical University from January 2008 to March 2011 were prospectively investigated.Identified by transesophageal echocardiography,patients were divided into two groups with respect to outcome:PFO group and non-PFO group.The recurrence of cerebral ischemic events was compared between the two groups after neurological follow-up.Results A total of 91 patients were recruited,including 57 patients with PFO and 34 patients without PFO.The follow-up period of two groups was 695 (506,1142) d.The recurrence rate at 15 months in patients with PFO (24.5％ (12/49)) was higher than those without PFO (6.9％ (2/29),x2 =4.391,P =0.036).Cum hazard curve indicated that recurrence risk of cerebral ischemic events in patients with CICVD in PFO group was higher than that of patients in non-PFO group during the follow-up period (P =0.044).Cox model used for multivariate survival analysis indicated that PFO was a risk factor for cerebral ischemic event recurrence among patients with CICVD (OR =4.159,95％ CI 1.178-14.689,P =0.027).Conclusions PFO is associated with increased recurrence risk of cerebral ischemia in CICVD patients.In addition,PFO may be a significant factor for ischemic cerebrovascular disease.

Adult ; Cardiomyopathy, Hypertrophic ; complications ; pathology ; Humans ; Male ; Myocardium ; pathology

Objective To establish a malignant pleural effusion model of Lewis lung cancer in C57BL/6 mice based on Micro Echo technology. Methods Single tumor cell suspension of Lewis lung cancer was injected into thoracic cavity.The pleural effusion was detected by Micro Echo technology.The volumes of effusion were quantified and the tumor cells were counted.Results After implanted of tumor cell, malignant pleural effusion can be detected by Micro Echo technology and observed after autopsy.Chemotherapy drugs such as Cyclophosphamide and Cisplatin can decrease the effusion volumes.Conclusion Pleural effusion model of Lewis lung cancer based on Micro Echo technology can be used to evaluate the efficacy of antitumor drugs.

Objective To evaluate the application value of T cell infected with Mybacterium tuberculosis assay in diagnosis of smear and culture negative pulmonary tuberculosis .Methods In total ,145 patients with smear and culture negative pulmonary tu‐berculosis and 45 patients with non‐tuberculosis lung disease were enrolled in the study .All patients received T cell infected with Mybacterium tuberculosis test and sensitivity ,specificity ,positive predicted value ,negative predicted value of testing for the diagno‐sis of smear and culture‐negative TB patients were calculated .Results The sensitivity of specificity T cell infected with Mybacteri‐um tuberculosis assay in diagnosis of smear and culture negative pulmonary tuberculosis was 85 .5% ,the specificity was 84 .4% , positive predicted value was 94 .7% ,negative predicted value was 64 .4% .No statistical significance in age‐dependent groups(P>0 .05) .Conclusion T cell infected with Mybacterium tuberculosis assay has high positive predicted value in diagnosis of smear and culture negative pulmonary tuberculosis and is suitable for clinical auxiliary diagnosis .

Objective To evaluate the value of diffusion-weighted imaging (DWI)in diagnosing the acute centrum ovale infarction, and also to investigate the pathogenesis of the infarction. Methods All 58 patients underwent conventional MRI and DWI scanning after symptoms’ onset. DWI findings were compared to the findings of T_1WI and T_2WI. Results The sensitivity and specificity in diagnosing the ischemia stroke were 96.4% and 98.8% within 7 days after onset. Of all the cases, 62.1% were associated with the cerebral large-vessel disease and emboligenic heart disease. Only 36.2% had a classic lacunar syndrome but 69.0% had more frequently an abrupt onset of symptoms. Conclusion DWI is of high accuracy for diagnosing centrum ovale infarction and detecting early infarction lesions which are difficult to be displayed in conventional MRI, and very helpful in differentiating the acute from non-acute lesions; symptomatic centrum ovale infarction is suggested to be associated with large-vessel and heart disease which should be distinguished from the lacunar infarcts.

Objective To analyze the risk factors for patients with cryptogenic ischemic cerebrovascular disease (CICVD) and patent foramen ovale (PFO), as well as to evaluate the relationship between common risk factors and PFO in cerebral ischemia. Methods Consecutive patients with CICVD aged 15 to 70 years who referred to Department of Neurology, Xuanwu Hospital, Capital Medical University from January 2008 to July 2011 were investigated. Identified by transesophageal echocardiography, they were divided into PFO group and non-PFO group with respect to outcome. The common risk factors of cerebral ischemic between 2 groups were compared. The relationship between these risk factors and PFO was analyzed. Results A total of 102 patients were investigated, including 61 patients (59.80%) with PFO and 41 patients (40.20%) without PFO. Positive family history of ischemic cerebrovascular disease proportion in PFO group (31.1%)was higher than that in non-PFO group (9.8%) (P=0.011). There was no significant difference in other observed indicators (P>0.05). Positive family history of ischemic cerebrovascular disease correlated with PFO among CICVD patients (r=0.251, P=0.011). Conclusion PFO was not only more common in CICVD patients, but also correlated with positive family history of ischemic cerebrovascular disease.

OBJECTIVE CYP2 family including CYP2C and CYP2J is the predominant arachidonic acid (AA) epoxygenase, and the epoxidation of AA produces four regioisomeric cis-epoxyeicosatrienoic acids (5,6-, 8,9-, 11,12-, and 14,15-EET). Human CYP2J2 is one of the main CYP isoforms expressed in brain, but CYP2C8 was present at a low level. The aim of this study is to investigate the roles of brain CYP2J in Parkinson disease. METHODS Rats received the right-unilaterally injection with concentrated LV-CYP2J3 or LV-EGFP in the substantia nigra (SN) at 3 d before LPS or 6-OHDA treatment. The animals were tested for rotational behavior with the dopaminergic agonist apomorphine dissolved in sterile saline at 14 and 21 d after LPS injection. The influence of CYP2J-dependent derivative, 14,15-EET, on the genes related with oxidative stress was assayed in SH-SY5Y cells. RESULTS CYP2J overexpression or 14,15-EET treatment significantly increased the levels of SOD1, CAT, GPX1, NRF2 and KEAP1 in neurons. TLR4- MyD88 signaling pathway was involved the down- regulation of CYP2J by LPS. The binding of p-CREB with the promoter of CYP2J was inhibited by the LPS treatment. The loss of dopami?nergic neurons in the right SN induced by LPS or 6- OHDA was significantly decreased by CYP2J3 transfection at 21 d after LPS injection. Compared with LPS or 6-OHDA group, the number of the rotation of rats was decreased by 42.6% and 60.7% by CYP2J3 transfection at 14 d after LPS or 6-OHDA injection;meanwhile, the rotation number was decreased by 12.7% and 21.3% at 21 d. The accumulation of alpha synuclein induced by LPS was significantly decreased by CYP2J3 transfection. The mRNA levels of SOD1, CAT, GPX1, NRF2 and KEAP1 in SN were decreased by LPS, which was attenuated by the injection of LV-CYP2J3. CONCLUSION Brain CYP2J can play a protective role in the damage of the inflammation and oxidative stress to the dopaminergic neurons. Brain CYP2J- dependent derivatives from AA may have therapeutic effects in Parkinson disease via the up- regulation of the antioxidant system in neurons.