The lack of effective in vitro infection model for hepatitis E virus (HEV) has greatly hindered the quantitative analysis of neutralizing titers of anti-HEV antibodies and human sera, thus impeding further studies of HEV-stimulated antibody responses and the immunological mechanisms. In order to improve this situation, the infection of HepG2 cells that are inefficient for HEV replication was continuously monitored until the viral load reached the limit of detection on day 13, the results of which confirmed the feasibility of using this cell line to establish the infection model. Then, neutralization assays of five anti-HEV murine monoclonal antibodies and serum samples collected from four HEV vaccine recipients (collected before and after vaccination) were performed by 96 multi-channel parallel infections, nucleic acid extraction, and qPCR. The results showed that the cell model can be applied for quantitative evaluation of the neutralizing capacity of different antibodies and antiserum samples from HEV vaccine recipients. In this study, we have successfully established a high-throughput in vitro HEV replication model, which will prove to be useful for the evaluation of HEV vaccines and studies of HEV epitopes.
Animals ; Antibodies, Viral ; analysis ; immunology ; Hepatitis Antibodies ; analysis ; immunology ; Hepatitis E ; immunology ; virology ; Hepatitis E virus ; chemistry ; immunology ; physiology ; High-Throughput Screening Assays ; methods ; Humans ; Mice ; Mice, Inbred BALB C ; Neutralization Tests ; methods ; Virus Replication

Objective To understand the mental authorization status and job burnout for domestic nursesandanalyze the correlation between them, and to explore new strategies to prevent and reduce nurse job burnout.Methods There were 203 clinical nursesinvolved, who were chosen from 5 first-class leveltop first class hospitals in Guangzhou, using mental authorized scale and job burnout scale questionnaires made by ourselves to investigate the basic information, psychological empowerment and job burnout.Results Perception of nurses’ mental authorizationachieved (4.75 ±0.67) (the medium level), while self-efficacy acquired the highest score (5.34 ±0.73) compared work effectiongained the lowest one (3.64 ±0.11).The nurses had suffered severe burnout, which performed asloss of personal accomplishment dimension (3.83 ±0.62), emotional exhaustion dimension (2.96 ±0.81) and depersonalization tendency dimensions (1.89 ±0.84);nurses perceived psychological empowerment and job burnout had negative correlation with emotional exhaustion, depersonalization tendencies, personal accomplishment dimension ( r = -0.436, -0.366, 0.514, respectively;P<0.01).Working influence had less correlation with job burnout dimensions, especially with the dimension of depersonalization work and job burnout dimensions affect the correlation was not obvious, especially with the tendency to personify no correlation dimension (r=-0.094,P>0.05).Conclusions The mental authorizationof clinical nurses just acquires middle level;hence, clinical nursing managers should enhance nurse's mental authorizationto release job burnout.

Objective To establish a model of screening succinate semialdehyde dehydrogenase (SSADH)inhibitors with anti-epilepsy effects,then using this model to find out the effective components in Tall Gastrodia Tuber (Latin:Rhizoma Gastrodiae,pinyin:Tianma)and some analogues and to study their structure-activity relationship.Methods First,SSADH enzyme system fluid was prepared,and the correlation of activity and optical density of SSADH was evaluated using UV spectrophotometry.Then,af-ter the screening method of SSADH activity was established by optimization of reaction temperature,reac-tion time,concentration of NAD +,SSA and SSADH,and pH of buffer on the SSADH activity,p-hydroxybenzaldehyde (HBA),a positive drug recorded in literature,was applied to verify the model. And the structural analogues of HBA were measured and the mechanism was analyzed.Results The screening model was established successfully,proved by HBA test.The UV detective system and its buff-ers of SSADH activity were determined with reaction temperature at 37 ℃ for 30 min,and detect wave at 340 nm.The structure-activity of HBA on GABA-T was as the same as that of vanillic aldehyde,and—OH and —CHO of benzene ring were the essential groups for inhibition SSADH.Conclusion The model established in this paper can be used for high throughput screening SSADH inhibitors and may guide the study of analogues of SSADH inhibitors and their mechanism.

OBJECTIVE To study the effects of Zigui yichong formula on premature ovarian insufficiency （POI） in mice through glycolysis metabolic pathway. METHODS Eighty SPF C57BL/6N female mice were divided into normal group， model group， Zigui yichong formula group （14.175 g/kg）， Zigui yichong formula+2-deoxy-D-arabino-hexose （2-DG） group （Zigui yichong formula 14.175 g/kg + glycolysis inhibitor 2-DG 100 mg/kg）， with 20 mice in each group. Except for the normal group， POI model mice were induced by intraperitoneal administration of cyclophosphamide in the other groups. After the model was successfully established， each group was given corresponding drugs. HE staining was employed to observe the pathomorphological changes in ovarian tissue and to count follicles at all developmental stages； radioimmunoassay was conducted to measure the serum levels of estradiol （E2）， anti-Müllerian hormone （AMH）， and follicle-stimulating hormone （FSH）； TUNEL assay was employed to detect apoptosis in ovarian granulosa cells of mice； the activities of hexokinase （HK）， pyruvate kinase （PK） and lactate dehydrogenase （LDH） were detected by colorimetry； Western blot and real-time fluorescence quantitative PCR were employed to analyze the protein and mRNA expressions of B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， caspase-3， HK2， pyruvate kinase M2 （PKM2）， and lactate dehydrogenase A （LDHA）. RESULTS Compared with model group， the number of primordial follicles， growing follicles， antral follicles and granulosa cells were increased significantly（P＜0.05）， and granulosa cells arranged neatly， but the number of atretic follicles and granulosa cells apoptosis were decreased significantly in Zigui yichong formula group （P＜0.05）； the serum levels of E2 and AMH， the activities of HK， PK and LDH， protein and mRNA expressions of Bcl-2， HK2， PKM2 and LDHA were increased significantly （P＜0.05）； the serum levels of FSH， the protein and mRNA expressions of Bax and caspase-3， Bax/Bcl-2 ratio were decreased significantly （P＜0.05）. 2-DG could reverse the improvement effects of Zigui yichong formula on the above indexes of POI model mice. CONCLUSIONS Zigui yichong formula may inhibit the apoptosis of ovarian granulosa cells， reduce follicle atresia and improve ovarian reserve function by promoting glycolysis levels in POI model mice.

The limited penetration of nanoparticles and their poor accessibility to cancer cell fractions in tumor remain essential challenges for effective anticancer therapy. Herein, we designed a targeting peptide-decorated biomimetic lipoprotein (termed as BL-RD) to enable their deep penetration and efficient accessibility to cancer cell fractions in a tumor, thereby improving the combinational chemo-photodynamic therapy of triple negative breast cancer. BL-RD was composed of phospholipids, apolipoprotein A1 mimetic peptide (PK22), targeting peptide-conjugated cytotoxic mertansine (RM) and photodynamic agents of DiIC18(5) (DiD). The counterpart biomimetic lipoprotein system without RM (termed as BL-D) was fabricated as control. Both BL-D and BL-RD were nanometer-sized particles with a mean diameter of less than 30 nm and could be efficiently internalized by cancer cells. After intravenous injection, they can be specifically accumulated at tumor sites. When comparing to the counterpart BL-D, BL-RD displayed superior capability to permeate across the tumor mass, extravasate from tumor vasculature to distant regions and efficiently access the cancer cell fractions in a solid tumor, thus producing noticeable depression of the tumor growth. Taken together, BL-RD can be a promising delivery nanoplatform with prominent tumor-penetrating and cancer cells-accessing capability for effective tumor therapy.