Inhibition of hepatitis B virus replication by APOBEC3G in vitro and in vivo

AIM： To investigate the effect of APOBEC3G mediated antiviral activity against hepatitis B virus （HBV） in cell cultures and replication competent HBV vector-based mouse model. METHODS： The mammalian hepatoma cells Huh7 and HepG2 were cotransfected with various amounts of CMV-driven expression vector encoding APOBEC3G and replication competent 1.3 fold over-length HBV. Levels of HBsAg and HBeAg in the media of the transfected cells were determined by ELISA. The expression of HBcAg in transfected cells was detected by western blot. HBV DNA and RNA from intracellular core particles were examined by Northern and Southern blot analyses. To assess activity of the APOBEC3G in vivo, an HBV vector-based model was used in which APOBEC3G and the HBV vector were co-delivered via high-volume tail vein injection. Levels of HBsAg and HBV DNA in the sera of mice as well as HBV core-associated RNA in the liver of mice were determined by ELISA and quantitative PCR analysis respectively. RESULTS： There was a dose dependent decrease in the levels of intracellular core-associated HBV DNA and extracellular production of HBsAg and HBeAg. The levels of intracellular core-associated viral RNA also decreased, but the expression of HBcAg in transfected cells showed almost no change. Consistent with in vitro results, levels of HBsAg in the sera of mice were dramatically decreased. More than 1.5 log10 decrease in levels of serum HBV DNA and liver HBV RNA were observed in the APOBEC3G-treated groups compared with the control groups.CONCLUSION： These findings indicate that APOBEC3G could suppress HBV replication and antigen expression both in vivo and in vitro, promising an advance in treatment of HBV infection.

Development Path and Urgency of Further Strengthening Construction of Public Hospitals Based on Novel Coronavirus Pneumonia Treatment

During the outbreak of novel coronavirus pneumonia(NCP)in Wuhan,public hospitals have played an important role in intensive care,case guidance and scheme optimization.At the same time,it also faces unprecedented challenges and tests.Based on the treatment of severe patients in Wuhan,combined with the treatment practice in Tongji Hospital affiliated to Tongji Medical College,Huazhong University of Science and Technology,this paper puts forward the urgency of further strengthening the construction of public hospitals,discusses the feasible path for promoting the development of public hospitals,so as to meet the growing medical needs of the people,improve the ability to respond to major public health emergencies,and effectively guarantee the safety of people's lives and the promotion of a healthy China construction.

Effects of PDTC on the Proliferation and PCNA Expression of Human Retinal Pigment Epithelial Cells

To investigate the effects of pyrrolidine dithiocarbamate （PDTC） on the proliferation and PCNA （proliferating cell nuclear antigen） expression of cultured human retinal pigment epithelium cells, human retinal pigment epithelium cells （RPE） were cultured from normal adults who died accidentally. The effects of PDTC on the proliferation of RPE cells were examined by using methyl thiazlyl tetrazolium （MTT） assay. The effects of PDTC on the PCNA expression of RPE cells were immunohistochemically examined by employing biological image analysis system （BIAS）. After treatment with PDTC of various of concentration ranging from 0.062 to 1 g/L for 24 h, or concentrations ranging from 0.031 to 1 g/L, the proliferation of RPE cells decreased in a dose-dependent manner. After treatment with PDTC of concentration varying from 0. 062 to 1 g/L for 24 h, the PCNA expression was also suppressed in a dose-dependent manner. It is concluded that PDTC can inhibit the proliferation of RPE cells in vitro in a dose-and time-dependent manner, at least in part, by down-regulating the expression of PCNA. PDTC may be used to prevent and treat the proliferative vitreoretinopathy （PVR）.

Analysis of Etiology and Drug Resistance of Biliary Infections

Summary: The bile was collected from fro patients with biliary infections, with the bacterium isolated to study the sensitivity of each kind of the bacterium to several antibiotics in common use. Except G-bacterium, we also found some kinds of G+ bacterium in infection bile. G-bacterium were not sensitive to Clindamycin, G+ bacterium were sensitive to Ciprofloxacin. Escherichia coli, Xanthomonas maltophilia, Enterobacter cloacae, Pseudomonas aeruginosa were sensitive to Ampicillin. G+ bacterium were not sensitive to Azactam. Enterococcus faecalis, Enterococcus faecium, Enterobacter cloacae were not sensitive to Ceftazidime. Enterococcus faecalis, Staphylococcus coagulase negative, Staphylococcus epidermidis, Pseudomonas aeruginosa were not sensitive to Ceftriaxone Sodium. We didn't found any bacterium resistance Imipenem. The possibility of the existence of G+ bacterium as well as drug resistance should be considered n patients with biliary infections. The value of susceptibility test should be respected to avoid drug abuse of antibiotics.

Inhibitory Effect of Tissue Transglutaminase (tTG) Antisense Oligodeoxynucleotides on tTG Expression in Cultured Bovine Trabecular Meshwork Cells

To study the effect of tTG fully phosphorothioated antisense oligodeoxynucleotides （tTG-ASDON） on tTG expression in cultured bovine trabecular meshwork cells （BTMCs） in vitro and explore a new treatment alternative for primary open angle glaucoma （POAG）, the ASDON1 and ASDON2 complementary to the protein codogram region of tTG were designed, synthesized and phosphorothioated according to the secondary structure of tTG. The ASDON1 and ASDON2 were embedded in Lipofectamine and transfected into BTMCs. The untreated group served as negative controls. The expression of tTG in the mRNA and protein level were measured by semi-quantitative RT-PCR and immunohistochemical technique-Supervision method respectively. Our results showed that both the mRNA and the protein of tTG with tTG-ASDON1 and tTCr-ASDON2 were significantly decreased as compared with that of the controls （P〈0.05）. On the other hand, no significant difference was found between the ASDON1 group and the ASDON2 group. It is concluded that the expression of tTG mRNA and protein in cultured BTMC are down-regulated by tTG- ASDON. As a result, tTG-ASDON may be used for the treatment of POAG through the inhibitory effect on the expression of tTG.

POLYMER-SUPPORTED β-CYCLODEXTRIN CATALYST FOR SELECTIVE SYNTHESISOF 4-H YDROXYBENZOIC ACID

4-hydroxybenzoic acid was synthesized by one pot method using polymer-bonded β-aminopropionic acid-2-β-cyclodextrin as a catalyst.by which the selectivity for 4-hydroxybenzoic acid is 87%. A possible carboxylation mechanism in the presence of the title catalyst is discussed. Also.the recovery and reusage of the catalyst is studied.

Effects of Electroacupuncture Combined with Repetitive Transcranial Magnetic Stimulation on the Expression of Nestin in Neural Stem Cell after Focal Cerebral Ischemia in Adult Rats

Objective:To investigate the influence of electroacupuncture(EA) combined with repetitive transcranial magnetic stimulation(rTMS) on the temporal profile of nestin expression after induction of focal cerebral ischemia in adult rats and to explore the mechanism of EA combined with rTMS in treating ischemic brain injury.Method:The model of transient focal ischemia was produced by occlusion of middle cerebral artery.Seventy-five Wistar rats were randomly divided into normal group,model group,EA group,rTMS group and EA +rTMS group.The neurologic impairment rating and ability of learning and memory were observed at the 7th、14th and 28th d after infarction respectively.Meanwhile,Western blotting was used to observe the number of nestin expression positive cells.Result:Nestin-positive cells were found in cortex,subgranular zone(SGZ),subventricular zone(SVZ) of the ipsilateral side at different time points after cerebral ischemia.The number of nestin-positive cells peaked at the 7th d,began to decrease at the 14th d and was significantly higher in EA+rTMS group than that in model group(P< 0.05),then almost reached normal at the 28th d.The improvement of neural motor function deficits as well as the indexes of learning and memory were more obvious in EA+rTMS group compared with model group(P<0.01,P<0.05).These effects were most obvious in EA +rTMS group compared with the EA and rTMS group(P<0.05).Conclusion:EA and rTMS possess the potency of building up and can increase the number of nestin-positive cells in some brain regions after focal cerebral ischemia,which might be one of the important mechanisms of EA combined with rTMS in treating ischemia brain injury.

Spontaneous magnetic field multipolar structure in toroidal plasmas based on 2D equilibrium

The multipolar magnetic field structure is investigated by the momentum conservation equation with self-c on sistent 3D sheared flows during transition of plasma properties from local paramagnetic to diamagnelic fields.Numerical results show that the traditional poloidal magnetic field(Bp)is one part of equilibrium magnetic fields.The non-zero-order quantities are originaled from the higher-order terms of 2D equilibrium treatment based on a Fourier expansion of 0(r,0).The distributions of magnetic field vectors of the order of 1,2,and 3 terms are presented respectively in two.four,and six polar fields with the local vortex structures(spontaneous magnetic conn ection).The excitation mechanisms of the magnetic vortices are the coupling effects of the magnetic fluid structure pattern and the toroidal effects.These results can help us understand the physical mechanism of the in teraction between the external perturbation fields and control tearing modes,as well as the radial plasma flow and magnetic vortices.

KLF6mRNA Expression in Primary Hepatocellular Carcinoma

Summary: To investigate the expression of KLF6mRNA in primary hepatocellular carcinoma (HCC), nomal liver tissues and the tissues adjacent to the cancers, reverse-transcription polymerase chain reaction (RT-PCR) was employed to investigate the expression of the KLF6 gene in HCC, the corresponding adjacent non-cancerous tissues and normal liver tissue. Our results showed that an amplified fragment of 427 bp DNA was detected in 18 of 19 (94.7 %) adjacent non-cancerous tissues and normal liver tissue, and in 12 (85.7 %) of 14 HCC. There were no significant differences in the levels of KLF6 mRNA between normal liver and liver tumors (P>0.05). It is concluded that KLF6 mRNA is generally expressed in HCC.

Long-term follow-up of haploidentical transplantation in relapsed/refractory severe aplastic anemia:a multicenter prospective study

In recent decades,haploidentical stem cell transplantation(haplo-SCT)to treat severe aplastic anemia(SAA)has achieved remarkable progress.However,long-term results are still lacking.We conducted a multicenter prospective study involving SAA patients who underwent haplo-SCT as salvage therapy.Long-term outcomes were assessed,mainly focusing on survival and quality of life(QoL).Longitudinal QoL was prospectively evaluated during pretransplantation and at 3 and 5 years posttransplantation using the SF-36 scale in adults and the PedsQL 4.0 scale in children.A total of 287 SAA patients were enrolled,and the median follow-up was 4.56 years(range,3.01–9.05 years)among surviving patients.During the long-term follow-up,268 of 275 evaluable patients(97.5%)obtained sustained full donor chimerism,and 93.4%had complete hematopoietic recovery.The estimated overall survival and failure-free survival for the whole cohort at 9 years were 85.4%±2.1%and 84.0%±2.2%,respectively.Age(≥18 years)and a poorer performance status(ECOG>1)were identified as risk factors for survival outcomes.For Qo L recovery after haplo-SCT,we found that QoL progressively improved from pretransplantation to the 3-year and 5-year time points with statistical significance.The occurrence of chronic graft versus host disease was a risk factor predicting poorer QoL scores in both the child and adult cohorts.At the last followup,74.0%of children and 72.9%of adults returned to normal school or work.These inspiring long-term outcomes suggest that salvage transplantation with haploidentical donors can be routine practice for SAA patients without human leukocyte antigen(HLA)-matched donors.

A GENERAL CLASS OF ONE-STEP APPROXIMATION FOR INDEX-1 STOCHASTIC DELAY-DIFFERENTIAL-ALGEBRAIC *EQUATIONS

This paper develops a class of general one-step discretization methods for solving the index-1 stochastic delay differential-algebraic equations. The existence and uniqueness theorem of strong solutions of index-1 equations is given. A strong convergence criterion of the methods is derived, which is applicable to a series of one-step stochastic numerical methods. Some specific numerical methods, such as the Euler-Maruyama method, stochastic ^-methods, split-step ^-methods are proposed, and their strong convergence results are given. Numerical experiments further illustrate the theoretical results.

A mathematic-physical approach to the satisfiability problem

A one-to-one and onto mapping between the set of conjunctive normal forms and a subset of the potential functions of static electric fields is given; it has been further proved that a conjunctive normal form is satisfiable if and only if there exists a zero point for its corresponding potential function. A particle is always moving in the direction of gradient descent in the field which is the fastest decreasing direction of potential of the particle. Thus, if a conjunctive normal form is satisfiable, the gradient method for its corresponding potential function becomes a fast algorithm to solve its satisfiability problem.

Integrated ionic sieving channels from engineering ordered monolayer two-dimensional crystallite structures

Atomically thin solid-state channels enabling selective molecular transport could potentially be used in a variety of separation and energy conversion applications.The density of channels,their height,distance and edge structure are the key factors that dramatically impact the selective transport performance.However,such channels with small constrictions and atomic precision have been limited to proof-ofconcept demonstrations based on microscale two-dimensional(2D)crystal stripes.Here,we report the engineering of highly ordered,scalable monolayer graphene crystallite arrays by chemical vapor deposition(CVD)method with a modified anisotropic etching approach.The size,shape,distance and edge structure of the graphene crystallite arrays in a large area could be delicately controlled through tailoring the synthetic parameters.This array structure can act as pillars to prop up a smooth single-crystal graphene film,and the fabricated integrated angstrom-size(3.4A)channels allow water transport but exclude hydrated ions,demonstrating potential in selective ionic sieving and nanofiltration practice.