To examine the effect of Gankang Suppository on duck hepatitis B virus (DHBV), the serum biochemistry and hepatic histology in an animal model of DHBV infection, a model of DHBV infection was established by infecting 1-day-old Yingtaogu ducklings with DHBV-positive serum. The successful model was confirmed by PCR assay and 48 ducklings infected with DHBV were randomly divided into 3 groups: a Gankang Suppository treatment group, an acyclovir (ACV) group and a DHBV model group (control), with each group having 16 animals. All the animals were given the medicines for 4 weeks in a row. The serum of the animals was taken 14 and 28 days after the medication and 7 days after drug discontinuation. Real-time PCR was performed to detect the copy numbers of DHBV DNA in the serum. ALT and AST were dynamically monitored. The ducklings were sacrificed on the 7th day after the discontinuation of the treatment and livers were harvested and examined for inflammation and degeneration of liver cells by using HE staining. The results showed that on day 14, 28 after the treatment and day 7 after the withdrawal, the logarithmic values (log) of DHBV DNA copy numbers in ducklings of Gankang Suppository treatment group were significantly lower than that before the treatment (P=0.0092, P=0.0070, P=0.0080, respectively). Compared with DHBV model control group, the ALT level was significantly decreased (P=0.0020, P=0.0019, respectively) on day 28 after the treatment and on day 7 after the withdrawal. The AST level was also reduced on day 14 after the treatment (P=0.0298). Compared with the ACV control group, the level of ALT was lower on day 7 after the withdrawal (P=0.0016). Histologically, the hepatocyte swelling, vacuolous degeneration and acidophilic degeneration in Gankang Suppository treatment group were alleviated 7 days after the withdrawal as compared with model control group (P=0.0282, P=0.0084, P=0.0195, respectively). It is concluded that Gankang Suppository can effectively suppress DHBV replication, reduce the levels of serum ALT and AST and improve hepatic histology.

Objective To evaluate the imaging characteristic and the value of application of breath hold two dimensional fast imaging employing steady state acquisition(FIFSTA) sequence, for aortic dissection(AD) magnetic resonance many time phases imaging. Methods To analyze retrospectively the manifestation of 5 cases with aortie dissection diagnosed, Bakey type Ⅰ,Ⅱ, Ⅲ, in 1, 1 and 3, all were examined using ECG triggered, two dimensional FIESTA sequence during end-expiratory breath hold. Scan with the long axes of aortic position(oblique position parallel with aortic arch) and transitional position. The results were analyzed by the soft of workstation AW4.2. Results The display rate of the true lumen and the false lumen of AD were 100% ,the display rate of membrane in aorta was 100% ,4 cases showed the hole of intimal tear,2 cases with thrombus,2 cases with aortic regurgitation,2 cases with ejecting in the hole of intimal tear. Conclusion Need not any contrast media, quickly acquired imaging, high contrast to noise ratio, least artifact,and especially show membrane in aorta and for the hole of intimal tear of aortic dissection(AD) magnetic resonance many time phases imaging with two dimensional FIESTA sequence.

Objective To investigate the clinical efficacy and safety of recombinant human B-type natriuretic peptide(rhBNP)in treatreat of patients with acute myocardial infarction(AMI).Methods 80 patients with AMI were randomly divided into control and treatment group.The two groups were accepted the standard foundation treatment(ABCDE)defined by the WHO and PCI therapy.The treatment group was added the rhBNP on the basis.The rhBNP was uniformly injected by intravenous with 1.5 μg/kg in 90s,and then with 0.0075 μg · kg-1 · min-1 in 48h; To investigate the BNP and myocardiolysis maker(Troponin I,cTnI)levels in blood-serum before and after the treatment;To evaluate left cardiac functions and left ventricular remodeling by left ventricular end-diastolic volume(LVEDV),left ventricular end-systolic volume(LVESV)and left ventricular ejection fraction(LVEF);To investigate the major cardiovascular events after treatment within 30 days.Results RhBNP could significantly reduce the concentration of the BNP and cTnI in blood-serum after treating one week(all P ＜ 0.05); It could remarkably deflate LV EDV and LVESV,improve LVEF(allP ＜0.05); It could reduce the incidence of major cardiovascular events(P ＜0.05)within 30 days(P ＜ 0.05).Conclusion The rhBNP could improve left ventricular pressure-phore and volume-phore,improve ventricular remodeling,improve myocardial ischemia and reduce major cardiovascular events within 30 days.It was proved that the rhBNP could improve AMI clinical prognosis.

BACKGROUND:Recently, glucocorticoid-induced necrosis of femoral head has been much studied. However, the precise Wnt signaling pathway by which puerarin suppresses adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells remains unconfirmed. OBJECTIVE:To investigate the expression of Wnt signaling pathway related genes and the key factor protein,β-catenin, during adipogenic differentiation of glucocorticoid-induced rat bone marrow mesenchymal stem cells treated by puerarin. METHODS:The third generation of bone marrow mesenchymal stem cells were cultured with Dulbecco’s modified Eagle’s medium containing blank serum (blank control group), dexamethasone (hormone group), dexamethasone with puerarin low dose group, the middle dose group and high dose group. After 6 days of culture, in the above five groups, the expressions of Wnt/β-catenin signaling pathway members, Wnt10b mRNA, GSK3βmRNA,β-catenin mRNA, were detected using RT-PCR assay, and the expression ofβ-catenin protein was detected using western blot assay.
RESULTS AND CONCLUSION:Compared with the control group, the Wnt10b mRNA,β-catenin mRNA andβ-catenin protein expressions were significantly higher in puerarin groups, but GSK3βmRNA expression was significantly lower in the puerarin groups. These findings suggest that puerarin effects on inhibition of adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells probably are realized through the activation of Wnt/β-catenin signal pathway and regulation of the key factor Wnt10b mRNA, GSK3βmRNA,β-catenin mRNA, andβ-catenin protein expressions. The mechanism by which puerarin prevents glucocorticoid-induced necrosis of femoral head not only improves local microcirculation of the femoral head, but also relates to its inhibitory effects on adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells.

Aplastic anemia (AA) is a Tlymphocyte-mediated autoimmune disease. The research on AA was focused on three areas, which were the pathogenesis of immune dysfunction, hematopoietic stem cell damage and abnormal hematopoietic microenvironment. In recent years, more attentions have been paid on abnormal immune mechanisms in the blood. There are complex intracellular cytokine and signal transduction pathway of pathogenesis in AA. And T-bet/IFN-γ signaling pathway plays an important role in development and progression of AA. This article aimed to review T-bet/IFN-γ signaling pathways in AA.

Objective To investigate the influence of primary cultured neonatal rat hippocampal neurons caused by human cytomegalovirus (HCMV AD169) infection on intracellular calcium and its mechanism.Methods Twenty SPF SD rats born within 24 hours(10 cases of male and 10 cases of female) were assigned to establish the primary rat hippocampal neuronal monolayer cells; After cultured 8 days in vitro,the eligible cells were randomly divided into HCMV infection group,HCMV + MK-801 group,MK-801 group and control group,with 10 wells in each group.The fluorescence intensity values of the intracellular free calcium were detected after 24 hours of treatment with Fluo-3AM fluorescence staining.Results Inoculation of HCMV neurons after 24 h turned to round and swollen gradually,and 4days later,most of the cells disappeared; by immunohistochemistry in cultures of hippocampal neurons in HCMV,visible early proteins,brownish yellow granules,hematoxylin were found after being stained with brown pigment.The fluorescence intensity values of neuronal intracellular calcium (215.5 ± 14.9) in HCMV group was higher than that of control group (116.4 ± 5.9) (t =15.2,P ＜ 0.01),whilerise,that in MK-801 group (88.1 ± 4.5) was significantly lower than that of control group,with decreased rate of (24.0 ± 6.7) ％ (t =-9.3,P ＜ 0.01).The fluorescence intensity values of neuronal intracellular calcium in HCMV + MK-801 group (135.5 ± 8.6) was significantly decreased compared with that of HCMV group (215.5 ± 14.9),with decreased rate of (37.0 ± 3.4) ％ (t =11.3,P ＜ 0.01).Conclusions Intracellular calcium overload of cultured rat hippocampal neurons in vitro with HCMV AD16 strains infection can be detected.One of its main mechanisms is the N-methyl-D-aspartic acid receptor channel-mediated calcium influx.

Objective To analyze the clinical characteristics, short-term and long-term outcome, overall survival and prognosis of nasal-type,natural killer T-cell lymphoma(NK/TCL) patients. Methods Clinical data of 45 patients with nasal NK/T cell lymphoma from Jan 2001 to Dec 2008 were analyzed retrospectively. Results The complete remission rate was 53.3% (24/45)and the partial remission rate was 17. 8% (8/45), the overall response rate was 71.1% (32/45). At a median follow-up of 15 months, the 1-year,3-year and 5-year survival rates were 68. 3% ,57. 1% ,and 43. 7% respectively. Multivariate analysis showed that B symptoms was independent prognostic factors of NK/TCL( P = 0. 001 ). Conclusions NK/TCL showed specific characteristics, and the long-term survival rate is still poor. Investigation of optional treatment is needed.

AIM: To construct a human recombinant immunoglobulin library containing D-Dimer by using phage surface display technology. METHODS: Human immunoglobulin heavy chain and light chain genes were amplified respectively by RT-PCR from different human lymphocytes using family specific primers and signal sequences of immunoglobulin as half-nested PCR primers. The heavy chain and light chain PCR products were cloned into phagemid vector pComb3H and the human immunoglobulin recombination library was generated with helper phage VCSM13. RESULTS: A human combinatorial antibody library consisting of 2.8?10~8 in dependent clones was generated with a titer of 4.1?10~(17)PFU/L. The recombinant frequency of Fab genes was 46%. CONCLUSION: A human combinatorial antibody library was generated. It will be beneficial for selecting Fab antibodies of D dimer from the library.

AIM: To study the relationship between the disturbance of nitric oxide/endothelin-1(NO/ET-1) and hepatic ischemia/reperfusion(I/R) injury as well as the regulation of NO/ET-1 system by hepatic ischemic preconditioning(IPC). METHODS: The changes of NO/ET-1 system and their relationship with hepatic I/R injury were compared between I/R group and IPC+I/R group in a rat hepatic I/R model. Two hours after reperfusion, the liver tissues were detected by RT-PCR to see whether there was inducible nitric oxide synthase (iNOS) mRNA expression. RESULTS:In the acute phase of hepatic reperfusion, the ratio of NO/ET-1 was reduced, which was due to a significant reduction of NO - 2/NO - 3 (the metabolic product of NO) and significant elevation of ET-1 in the blood plasma. The content of ALT, AST, LDH and TNF-? in blood plasma, and of MDA in liver tissue were increased but ATP in liver tissue was reduced, the hepatic damage was deteriorated. The protection of the hepatic IPC was concerned with the elevation of the ratio of NO/ET-1 caused by the elevation of NO - 2/NO - 3, and reduction of ET-1 as well. There was no iNOS mRNA detected in the liver tissues.CONCLUSION: Hepatic I/R injury is related to the disturbance of NO/ET-1. The protection of the hepatic IPC in the acute phase might be conducted by its regulation of NO/ET-1 system. The cNOS rather than the iNOS generated the NO in this situation.

Objective:To evaluate the feasibility of two materials used as scaffolds in periodontal tissue engineering. Methods: Dog periodontal ligament cells cultured in vitro were collected and seeded on three-dimensional framework of cancellous bone matrix (CBM) or nano-HAp/collagen (nHAC) . The cell growth in the scaffolds was observed by cell counting and scanning electronic microscope. Results: Porous structure of the two materials and adhesion and eugonic growth of cells in the scaffolds were observed by scanning electronic microscope.The cell number was doubled in both scaffolds in 72 h culture. Conclusion: It may be feasible to use CBM or nHAC as the scaffolds for periodontal tissue engineering.