Objective To evaluate the inhibitory effect of the small interfering RNA(siRNA) on hepatitis B virus(HBV)in vivo which targets HBV S gene region.Methods An animal model of HBV infection was developed hydrodynamically by injecting pcDNA3.1-HBV together with siRNA through the tail vein of Balb/c.HBsAg was analyzed by time resolved immunofluorometric assay, HBV DNA was analyzed by fluorogenic quantitative PCR(FQ-PCR),HBV S-mRNA was detected by semi-quantitative RT-PCR,and viral specific proteins(HBsAg and HBcAg)in the liver were assayed by immunohistochemical staining.Results In the mice,the siRNA could effectively inhibit the secre- tion of HBsAg,reduce the titers of HBV DNA,and immunohistochemical results also indicated that the number of HBsAg and HBcAg positive cells was reduced.The inhibitory effect of siRNA on HBV lasted 3 clays at least.Conclusion These results demonstrate that the siRNA targeting HBV S gene region can substantially and specifically inhibit HBV replication and expression in vivo.

39℃)developed at the progressive stage of this disease.Physical examination showed variously sized,round or oval,atrophic and variola-like scars along with scattered erythematous patches,papules, necrosis and crusts on the face and extremities.The face was edematous,and there were some edematous and erythematous plaques with a necrotic center on the legs and arms.Histological examination revealed a massive infiltration with atypical CD8~+lymphocytes around the vessels and appendages in dermis.A diagnosis of CD8~+cutaneous T-cell lymphoma(CTCL)was made.Glucocorticoid and immunosuppressants were effective in controlling the condition.Up to the time of the writing,there has not been any definite evidence of systemic involvement.

Objectives To study the level and development of serum specific antibody against severe acute respiratory syndrome coronavirus(SARS-CoV)of different populations in SARS pestilence district after SARS outbreak in a general hospital.Discuss SARS sub-clinical infection and protective action of the IgG antibody.Methods Seroepidemiology method,enzyme-linked immunosorbent assay (ELISA)and indirect immunfluorescence assay(IFA)were employed to investigate the changing level of serum antibody to SARS-associated coronavirus in non-SARS population in SARS pestilence district during and after SARS outbreak.The development of IgM and IgG antibody in patients with SARS in 6 weeks after the onset of SARS was studied qualitatively.The level changing of IgG antibody in con- valescent patients with SARS in 82 weeks after the onset was observed dynamically.Results The ELISA test outcome of IgG antibody was negative in 200 non-SARS people who were random samples of normal mass in SARS pestilence district and common community.The positive rate was 0.41% in 487 SARS high risk population tested by ELISA,but showed negative when retested by IFA.The A value level of IgG antibody existed significant difference in non SARS mass during and after SARS outbreak and the later's was higher them the former's(P

Mean hemoglobin (Hb) concentration of about 3 500 subjects derived from 17 studies of Himalayan highlanders (Tibetans, Sherpas, and Ladakhis) was compared with lowlanders (Chinese Han, Indian Tamils) lived in the Himalayas, and European climbers during Everest expeditions as well as Andean natives. The results found that Hb concentration in Himalayan highlanders was systemically lower than those reported for Andean natives and lowland immigrants. These comparative data demonstrated that a healthy native population may successfully reside at high altitude without a significant elevation in Hb, and the lower Hb levels of Himalayan highlanders than those of migrated lowlanders and Andean natives are an example of favourable adaptation over the generations. In addition, excessive polycythemia has frequently been used as a marker of chronic mountain sickness (CMS). Altitude populations who have a higher Hb concentration also have a higher incidence of CMS. The low Hb in Himalayans suggested as showing adaptation over many generations in Tibetan stock. Recent work in Tibet, suggested that Tibetans there may have adapted to high altitude as a result of evolutionary pressure selecting for genes which give an advantage at altitude. All of the population genomic and statistical analysis indicated that EPAS1 and EGLN1 are mostly likely responsible for high altitude adaptation and closely related to low Hb concentration in Tibetans. These data supported the hypothesis that Himalayan highlanders have evolved a genetically different erythropoietic response to chronic hypoxia by virtue of their much longer exposure to high altitude.
Adaptation, Physiological ; Altitude ; Asian Continental Ancestry Group ; genetics ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Evolution, Molecular ; Hemoglobins ; genetics ; Humans ; Hypoxia-Inducible Factor-Proline Dioxygenases ; genetics ; Tibet

OBJECTIVETo observe MET-associated alteration during the trans-differentiation from MSCs to neuron-like cells, and to explore the possible molecular mechanism.

METHODSBone marrow MSCs were isolated from rat femur and purified in continuous cell culture. After induced differentiation to neuron-like cells by the combination of butylated hydroxyanisole (BHA) and dimethyl sulfoxide (DMSO), cells were tested by comparative polymerase chain reaction (PCR) for the relative expression of MET biomarkers and transcription factors, and for cell cycle by flow cytometry. Meanwhile, target genes of Wnt/β-catenin pathway were also analyzed by comparative PCR to determine the possible involvement.

RESULTSIn MSC-induced neuron-like cells, MET-associated transcription factors such as Snail, Slug, ZEB1, ZEB2, and Twist were significantly attenuated in expression level. The Mesenchymal marker Vimentin expression level was increased. Membrane protein E-cad was slightly down-regulated, while N-cad level was marginally elevated. Percentage of proliferating cells (S phase in cell cycle) markedly shrank from 40.42% for MSCs to 6.76% for MSC-derived neuron. Additionally, Wnt/β-catenin target genes β-catenin and c-myc were decreasingly expressed.

CONCLUSIONChemically induced trans-differentiation from MSC to neuron caused similar MET-featured alteration in gene expression and proliferation to known MET, which might be underlied by deactivation of Wnt/β-catenin pathway.

Animals ; Epithelial-Mesenchymal Transition ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism ; Rats ; Wnt Signaling Pathway ; beta Catenin ; metabolism

OBJECTIVE Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited disease with a high morbidity around 1/1000-1/400, characterized by progressive enlargement of fluid-filled cysts derived from renal tubular epithelial cells. Massive cysts gradually compress renal parenchyma destroying normal renal structures and compromising renal functions. Unfortunately, it will cause end-stage renal disease in most of the patients but without effective therapy now, who have to live on hemodialysis or kidney transplantation. Based on this present situation, it is of great significance to find early intervention to inhibit renal cyst development. The projective of this study was to investigate whether Ganoderma triterpenes (GT) can inhibit renal cyst development and study the related mechanism. METHODS and RESULTS First, we used MDCK cyst model, cultivated MDCK cells in vitro to form fluid-filled cysts surrounded by monolayer cells. GT inhibited MDCK cyst formation significantly, and inhibited cyst enlargement dose-dependently proving GT cyst inhibition in vitro. Then we used an embryonic kidney cyst model, wile-type mice kidneys were taken out on embryonic day 13.5 to form renal cysts stimulated with 8-Br-cAMP. GT inhibited embryonic kidney cyst development significantly in a dose-dependent and reversible manner proving GT cyst inhibition at organ level. Furthermore, we used two ADPKD mouse models with severe cystic kidney disease phenotypes. GT dramatically inhibited renal cyst development, decreased ADPKD mouse kidney volume and the cyst index inside proving GT cyst inhibition in vivo. By Western blot, we proved GT down-regulated Ras/MAPK signal pathway without detectable effect on mTOR signal pathway both in MDCK cells and two ADPKD mouse kidneys. CONCLUSION GT retard renal cyst development both in vitro and in vivo significantly. The related mechanisms were involved in GT promoting renal tubular epithelial cell differentiation, down-regulating intracellular cAMP level and Ras/MAPK signal pathway.

OBJECTIVETo study the diagnostic accuracy of hematolymphoid malignancy by using effusion fluid cytology specimens and to evaluate the values of immunocytochemistry for this assay.

METHODSThe cytospin preparations/smears and cell block sections of effusion cytology specimens from 33 cases of hematolymphoid malignancy were retrospectively reviewed. Immunocytochemical study was performed. In selected cases, in-situ hybridization for Epstein-Barr virus-encoded RNA and immunoglobulin and T-cell receptor gene rearrangement study were carried out as indicated.

RESULTSThere were 33 cases of hematolymphoid malignancy, including 12 cases of T-lymphoblastic leukemia/lymphoma, 16 cases of mature B cell neoplasm (including 9 cases of diffuse large B-cell lymphoma, 2 cases of Burkitt lymphoma, 2 cases of plasmacytoma/multiple myeloma, 2 cases of B-small lymphocytic leukemia/lymphoma and 1 case of mantle cell lymphoma), 3 cases of mature T or NK-cell neoplasm (including 1 case of extranodal nasal NK/T-cell lymphoma, 1 case of angioimmunoblastic T-cell lymphoma and 1 case of T-cell prolymphocytic leukemia), 1 case of myeloid sarcoma and 1 case of mast cell sarcoma. Amongst the 33 cases studied, 16 represented disease relapses, including 8 cases of diffuse large B-cell lymphoma, 2 cases of plasmacytoma/multiple myeloma, 2 cases of B-small lymphocytic leukemia/lymphoma, 1 case of T-lymphoblastic leukemia/lymphoma, 1 case of angioimmunoblastic T-cell lymphoma, 1 case of mantle cell lymphoma and 1 case of mast cell sarcoma. The remaining 17 cases showed serous effusion as the primary manifestation, with the diagnosis primarily made upon cytologic examination. The cytologic findings seen in all the 33 cases studied were in agreement with the corresponding histologic diagnosis.

CONCLUSIONSDiagnosis of hematolymphoid malignancy by effusion fluid cytology specimens is possible, especially when coupled with the clinical history, immunophenotype, in-situ hybridization and gene rearrangement study findings. This is especially so for cases with disease relapses.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Ascitic Fluid ; metabolism ; pathology ; Burkitt Lymphoma ; diagnosis ; metabolism ; pathology ; Child ; Cytodiagnosis ; methods ; Female ; Humans ; Immunohistochemistry ; Lymphoma, Extranodal NK-T-Cell ; diagnosis ; metabolism ; pathology ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; metabolism ; pathology ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; metabolism ; pathology ; Plasmacytoma ; diagnosis ; metabolism ; pathology ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; pathology ; Retrospective Studies ; Young Adult

Objective: To investigate the effect of different doses of perindopril on peripheral endothelial progenitor cells (EPCs) and vascular endothelial function in patients with coronary artery disease (CAD) .
Methods: A total of 84 CAD patients with coronary angiography confirmed diagnosis were divided into 3 groups: Control group, the patients received routine medication, n=27. Low-dose group, the patients received routine medication with perindopril for 4mg, n=29. High-dose group, the patients received routine medication with perindopril for 8mg, n=28. All patients were treated for 12 weeks. The EPCs level was detected by flow cytometry assay, flow-mediated-dilation (FMD) function in brachial artery was measured by ultrasound and plasma levels of high sensitivity C-reactive protein (hs-CRP), angiotensin II (AngII) were examined in all groups.
Results: ① After12 weeks of treatment, the EPCs level and FMD function had certain improvement, hs-CRP level decreased in various degrees in all 3 groups, P<0.05, and AngII level decreased in both perindopril groups, P<0.05.②After treatment, compared with Control group, both perindopril groups had the increased EPCs level and FMD function, while decreased levels of hs-CRP and AngII, P<0.01.③Compared with Low-dose group, High-dose
group showed increased EPCs level and FMD function, decreased levels of hs-CRP and AgnII, P<0.05.
Conclusion: Perindopril may mobilize peripheral EPCs at certain point, and therefore improve endothelial function, the higher dose of perindopril may have better effect.

OBJECTIVETo explore the podocyte injury in patients with diabetic nephropathy (DN) and analyze its relationship with glucose regulated protein 78 (GRP78) and proteinuria.

METHODSThe clinical data of 48 patients diagnosed as DN by renal biopsy were reviewed. All patients were divided into two groups according to proteinuria (>3.5 g/d, n=31 and 3.5 g/d, n=17). The density of podocytes was illustrated by immunohistochemistry staining of Wilms tumor-1 (WT-1), and the immunofluorescence double-staining results of synaptopodin and GRP78 in podocytes were detected.

RESULTSThe podocyte dentistry of urine protein > 3.5 g/d group was significantly lower than that of urine protein>3.5 g/d group urine protein<3.5 g/d group(P=0.003), and it was negatively correlated with proteinuria (P=0.005). The expressions of synaptopodin and GRP78 in podocytes were also negatively correlated with proteinuria (P=0.004 and P=0.001).

CONCLUSIONThe podocyte injury is aggravated with increased proteinuria in DN patients, along with the decrease of the adaptive ability of endoplasmic reticulum to stress.

Adult ; Diabetic Nephropathies ; complications ; metabolism ; pathology ; Female ; Heat-Shock Proteins ; metabolism ; Humans ; Male ; Middle Aged ; Podocytes ; pathology ; Proteinuria ; etiology

OBJECTIVETo study the clinical significance of changes of serum myocardial enzymes in patients with acute carbon monoxide poisoning.

METHODSTo determine the dynamic changes of the activity of myocardial enzymes and ECG in 62 patients with acute CO poisoning.

RESULTSIn patients with acute CO poisoning 5 kinds of myocardial enzymes begin to increase within 24 hours, the activities of aspartate aminotransferase (AST), creatine phosphokinase (CPK), lactic dehydrogenase (LDH), alpha-hydroxybutyrate dehydrogenase (alpha-HBDH), CPK isoenzyme (CK-MB) were (20.2 +/- 12.3), (151.6 +/- 91.8), (146.8 +/- 50.4), (154.8 +/- 47.7), (13.8 +/- 8.1) U/L respectively, while those in control group were (12.1 +/- 6.7), (90.6 +/- 17.3), (118.7 +/- 13.5), (89.9 +/- 27.9), (5.9 +/- 3.3) U/L respectively. There was significant difference between two groups (P < 0.01); 3 d later, the activities of 5 enzymes were still increased [(21.3 +/- 12.3), (105.8 +/- 51.4), (144.8 +/- 51.4), (159.8 +/- 35.4), (16.2 +/- 9.1) U/L respectively]. 7 and 12 d later, the activities of alpha-HBDH and CK-MB were still higher than those of control (P < 0.01). LDH(1) and LDH(2) increased to peak value in 24 h after poisoning (35.3 +/- 5.8), (43.8 +/- 5.7) U/L vs (24.8 +/- 3.9), (36.9 +/- 4.3) U/L, P < 0.01. The abnormal rate of serum LDH(1) was 78.7%, LDH(2) 58.3%, LDH 45.2%, CK-MB 37.1%, alpha-HBDH 33.6% and the abnormal rate of ECG was less than 10%.

CONCLUSIONAcute carbon monoxide poisoning may cause myocardial injury. Determination of serum myocardial enzymes may contribute to showing myocardial injury, early diagnosis and treatment, results of treatment and prognosis.

Adult ; Carbon Monoxide Poisoning ; blood ; enzymology ; Creatine Kinase ; blood ; Creatine Kinase, MB Form ; Female ; Humans ; Hydroxybutyrate Dehydrogenase ; blood ; Isoenzymes ; blood ; L-Lactate Dehydrogenase ; blood ; Male ; Middle Aged ; Myocardium ; enzymology