Enterovirus A71 (EV-A71) causes hand, foot, and mouth disease (HFMD) and various neurological complications, including aseptic meningitis and neurogenic pulmonary edema in young children. HFMD caused by EV-A71 have broken out several times in the Asia-Pacific region since 2007. And it has been a serious threat to public health. There is no effective vaccine or antiviral drug. The pathogenesis of EV-A71 infection is unknown, and EV-A71 3C protein plays an irreplaceable role in replication and anti - innate immunity. Further research on EV-A71 3C protein is conducive to understand the pathogenesis of EV-A71 infection and antiviral drug.
Animals ; Antiviral Agents ; pharmacology ; Enterovirus ; drug effects ; immunology ; metabolism ; physiology ; Humans ; Immunity, Innate ; Viral Proteins ; chemistry ; genetics ; metabolism ; Virus Replication ; drug effects

BACKGROUND:The early damaged chondrocytes are susceptible to de-differentiate and exert unstable phenotype during the in vitro culture, thus needing some growth factors.
OBJECTIVE:To observe the promotion effect of insulin-like growth factor 1 on the in vitro proliferation of chondrocytes in adult rabbits with traumatic arthritis.
METHODS:Traumatic arthritis models of adult rabbits were established by using the modified Hulth method. After the models were successful y established, the distal femur and proximal tibia were harvested under sterile conditions, the chondrocytes were cultured. The cultured cells were divided into two groups:control group was cultured with Dulbecco’s modified Eagle’s medium containing 10%fetal bovine serum, while experimental group was cultured with Dulbecco's modified Eagle’s medium containing 100μg/L insulin-like growth factor 1. The effect of insulin-like growth factor 1 on the proliferation of chondrocytes in adult rabbits with traumatic arthritis was determined through the cytomorphology, cellcounting, and cellactivity.
RESULTS AND CONCLUSION:The chondrocytes in adult rabbits with traumatic arthritis were successful y cultured, the majority of cells were mini-cells, presenting smal fusiform, round or polygonal shape. Hematoxylin-eosin staining showed that the number of cells in experimental group was higher than that in control group. MTT assay found that the absorbance of cells in experimental group was greater than that in control group (P<0.01). Our findings indicate that, insulin-like growth factor 1 can promote the in vitro proliferation of chondrocytes in adult rabbits with traumatic arthritis.

A sensitive, rapid and accurate liquid chromatography-tandem mass spectrometric (LC-MS/MS) method with pre-column derivatization was developed for the simultaneous determination of erdosteine and its thiol-containing active metabolite in human plasma. Paracetamol and captopril were chosen as the internal standard of erdosteine and its active metabolite, respectively. Aliquots of 100 microL plasma sample were derivatized by 2-bromine-3'-methoxy acetophenone, then separated on an Agilent XDB-C18 (50 mm x 4.6 mm ID, 1.8 microm) column using 0.1% formic acid methanol--0.1% formic acid 5 mmol x L(-1) ammonium acetate as mobile phase, in a gradient mode. Detection of erdosteine and its active metabolite were achieved by ESI MS/MS in the positive ion mode. The linear calibration curves for erdosteine and its active metabolite were obtained in the concentration ranges of 5-3 000 ng x mL(-1) and 5-10 000 ng x mL(-1), respectively. The lower limit of quantification of erdosteine and its active metabolite were both 5.00 ng x mL(-1). The pharmacokinetic results of erdosteine and its thiol-containing active metabolite showed that the area under curve (AUC) of the thiol-containing active metabolite was 6.2 times of that of erdosteine after a single oral dose of 600 mg erdosteine tables in 32 healthy volunteers, The mean residence time (MRT) of the thiol-containing active metabolite was (7.51 +/- 0.788) h, which provided a pharmacokinetic basis for the rational dosage regimen.

Objective To explore the diagnostic value of needle stereotaction marking by steel wire(NSMSW) under mammography for nonpalpable early stage breast carcinoma (BC). Methods 29 patients with nonpalpable breast lesions were performed NSMSW under mammography,and the lesions were removed for biopsy to make the diagnosis. Results Nine patients(31.0%) were definited as early stage BC and twenty(69.0%) benign disease.The pathologic results in freezing sections and in parafin sections were the same. Conclusions NSMSW under mammography can resolve the problem of accurate location of nonpalpable breast lesions during operation .It is a credible, useful and practical method for diagnosis of nonpalpable early stage BC.

Objective To explore the strategy of surgical therapy for breast carcinoma . Methods The clinical data of 258 patients with breast carcinoma were analysed retrospectively. Results (1)136 patients with stage Ⅰ～Ⅱ breast carcinoma were subjected to modified radical mastectomy, overall survival(OS) was 100%, and relapse free survival(RFS)92.6%.(2)Partial mastectomy and axillary dissection were performed on two patients with stage Ⅱbreast carcinoma,one relapsed in 5 months after operation. (3)In patients with stage Ⅲ breast carcinoma,there was no statistical difference in OS and RFS between 88 patients subjected to modified radical mastectomy and 20 radical mastectomy.(4)The radical operation showed a better efficacy in 5 patients with stage Ⅳ breast carcinoma.(5)Using special breast cutter and electrotome,the rate of surgical blood transfusion was 3.5%,postoperative hematocele 2.7%,flap necrosis 7.4%, effusion under skin 18.6%,and edema of affected limb 4.3%. Conclusions (1)The modified radical mastectomy is the major operation for stage Ⅰ～Ⅲ breast carcinoma patients. (2)Using special breast cutter and electrotome could cut down surgical blood transfusion and operation time.(3)Rational axillary lymph node dissective could reduce postoperative complications.

Aim To synthesize 8-bromo-ethoxy Rhein and investigate its mechanisms and inhibition effect on hepatitis B surface antigen ( HBsAg ) and e antigen ( HBeAg) in HepG2.2.15 cells.Methods 8-bromo-ethoxy Rhein was synthesized based on the chemical structure of Rhein , and its structure was identified by IR,1 H-NMR and 13 C-NMR spectra.MTT assay was used to test the inhibitory effect of 8-bromo-ethoxy Rhein on HepG2.2.15 cells.After the cells treatment by 8-bromo-ethoxy Rhein , the HBsAg and HBeAg in cell supernatant were detected by ELISA .The expres-sion of hepatitis B virus X gene ( HBx) was detected by Western blot .The cell cycles were examined with flow cytometry.The intracellular free calcium concentration was detected by laser scanning confocal microscopy . Results The structure of 8-bromo-ethoxy Rhein was confirmed by IR,1 H-NMR and 13 C-NMR.MTT results showed that synthetic product and Rhein could inhibit the cell proliferation in HepG2.2.15 cells.After trea-ted with 8-bromo-ethoxy Rhein and Rhein for 72 h,the half inhibitory concentration 50%( IC50 ) was 14.29 mg? L-1 and 11.59 mg? L-1 , respectively .Using non-cytotoxic dose of 8-bromo-ethoxy Rhein , the inhibitory effect on HBsAg and HBeAg was gradually enhanced with increasing 8-bromo-ethoxy Rhein concentration . The inhibitory effect of synthetic product on hepatitis B virus was better than that of Rhein .8-bromo-ethoxy Rhein could down-regulate the expression of HBx , in-tracellular calcium ion concentration and block the hepatitis B virus ( HBV ) replication.Flow cytometry results showed 8-bromo-ethoxy Rhein didn′t affect the cell cycle .Conclusions Compare with Rhein , the synthesis of 8-bromo-ethoxy Rhein shows stronger inhi-bition on hepatitis B virus in HepG2.2.15, and its mechanisms may involve down-regulating the expres-sion of HBx and reducing calcium ion concentration .

Objective To investigate the role of a small RNA interference against VEGF in radiation sensitivity in melanoma .Methods A375 human melanoma cell lines were transplanted into nude mice ,which with malignant melanoma were randomly divided into control group , VEGF negative plasmid group and VEGF positive plasmid group, followed by 4Gy irradiation twice a week for 2 weeks.The volume of tumor was calculated twice a week, the area of tumor necrosis was assayed by HE,the expression of VEGF in tumor was determined by Western-blot and Immunohistochemical. ResuIts The expression of VEGF in VEGF positive plasmid group decreased significantly (P<0.05), VEGF positive group had more tissue necrosis, tumor growth was significantly inhibited (P<0.05).ConcIusion siRNA-VEGF in tumor injection liposome encapsulated in malignant melanoma has a role in the radiation sensitization, which provides an experimental basis for the clinical development of targeted therapy combined with radiotherapy for the treatment of VEGF gene.

Objective To make comparisons of the three models of acute and chronic rheumatic carditis to find out an optimal animal model.Methods AntigenⅠwas a emulsifier mixed by complete freund’ s adjuvant( CFA) and Group A streptococcus(GAS).AntigenⅡwas mixed by incomplete freund’s adjuvant(IFA) and GAS.Female Lewis rats were randomly divided into four groups: A, B, C treatmeat groups were immuned with antigenⅠat the foot pad firstly. Subsequently, rats in group A、B、C were injected antigenⅠ, antigenⅡand activated GAS respectively to make the models of RHD.Rats in control group D were immunized with the same protocol outlined as treatment groups but without GAS. Respectively 7, 12, 24 weeks the rats were sacrificed 24 ( each group was 6).The blood biochemical item and Hematoxylin-eosin( HE) staining of hearts were detected.Results In group C the mortality was 25%.In group A, the incidence of carditis was the highest.Histopathological manifestations of group A, C was not only revealed acute damage such as inflammatory cell infiltrate as well as group B, but also the Aschofflike cells in the myocardial cells interstitial.But in group A and C there had a great degree of the inflammatory cells infiltration than group B.At 24th week rats in group A detected the rate and degree of valve fibrosis in chronic damage were higher than group B and C.None of rats in group D presented carditis or valvulitis.Conclusion In group A, giving the GAS with continuous stimulation after using the mixed emulsification of CFA and GAS to immune Lewis rats for five times was a appropriate method which could provide an optimal animal model for experimental study of acute and chronic rheumatic heart disease.

The clinical data of 4 children with congenital absence of tracheal cartilage ring in Qilu Children′s Hospital of Shandong University from November 2017 to August 2019 were retrospectively analyzed.The age of the 4 cases was from 2 months to 1 year.One case had no obvious symptoms after birth, and then had repeated wheezing attacks for 3 times.Another 3 cases had symptoms after birth, such as throat ringing, repeated cough and asthma.Bronchoscopy in 4 cases indicated that the local lumen of trachea was round and centripetal stenosis, but the body of bronchoscope could pass smoothly, and no cartilage structure was found in the wall of trachea.Among them, 3 cases of chest enhanced CT examination displayed local and hourglass-like tracheal stenosis, without vascular ring malformation.All the 4 cases were treated by operation, of which 3 cases recovered well and 1 case died of severe infection after operation.

Objective To construct a chimeric infectious clone of the fatal virulent strain SDLY 107, containing the gene fragments encoding 2A and 3B proteins of the mild virulent strain SDLY 1, and to establish a reverse genetic system platform for further investigation on virulence of enterovirus 71 strains. Methods The overlap PCR analysis was performed to obtain the gene fragments encoding 2A and 3B pro-teins of the mild virulent strain SDLY 1.The obtained gene fragments were digested and then cloned into a plasmid pMD19-T containing the full-length gene of SDLY 107 strain by using gene replacement strategy. The recombinant RNA was transfected into Vero cells for the preparation of recombinant virus particles.Sev-eral assays including the PCR, indirect immunofluorescence ( IFA) , Western blot and sequencing were per-formed for virus identification.Virus titers were measured by 50%cell culture infective dose ( CCID50 ) and plaque assay.Results The infectious clones of SDLY 107-2A-1 and SDLY 107-3B-1 chimeric virus strains were constructed successfully.Typical cytopathic effect was observed in Vero cells after viral transfection. Identification of the rescued viruses by PCR, IFA, Western blot and sequencing further confirmed the suc-cessful construction of infectious virus strains.The virus titers of SDLY 107-2A-1 and SDLY 107-3B-1 strains detected by CCID50 and plaque assay were 1.25 ×105 PFU/ml and 0.7 ×105 PFU/ml, respectively. Conclusion The chimeric viruses SDLY 107-2A-1 and SDLY 107-3B-1 were rescued successfully, causing cytopathic effects similar to those by using the parental virus strain SDLY 107.This study might pave the way for further investigation on in vitro and in vivo virulence of enterovirus 71 strains.