To identify the active constituents in vitro and blood-absorbed ingredients in vivo from Yin Chen Hao decoction provides scientific evidence for probing its prevention and treatment mechanism on acute liver injury. An ultrahigh performance liquid chromatography quadrupole-time of flight-mass spectrometry (UPLC-QTOF/MS) method was applied for analysis of Yin Chen Hao decoction and the serum samples of mice with con-A induced acute liver injury after preventive oral administration for 14 days (the use of all laboratory animals in this study was approved by the Ethics Committee of the Naval Medical University, 19YF1459400). A total of 90 chemical constituents were identified from Yin Chen Hao decoction, mainly were flavonoids, terpenoids, tannins, quinones. 5 prototype compounds were identified in the serum, including chrysophanol, deoxyrhapontin-8-O-gallate, mussaenosidic acid, herniarin, emodin. The established UPLC-QTOF/MS method could efficiently and sensitively identify the constituents in vitro and blood-absorbed ingredients of Yin Chen Hao decoction, primarily clarify the material basis of its hepatoprotective effect, and provided a scientific basis for the quality marker selection and the pharmacodynamic material basis research on the decoction.

BACKGROUNDUrethral reconstruction for both congenital and acquired etiologies remains a challenge for most urologic surgeons. Tissue engineering has been proposed as a strategy for urethral reconstruction. The purpose of this study was to determine whether a naturally derived extracellular matrix substitute developed for urethral reconstruction would be suitable for urethral repair in an animal model.

METHODSA urethral segmental defect was created in 20 male rabbits. The urethral extracellular matrix, obtained and processed from rabbit urethral tissue, was trimmed and transplanted to repair the urethral defect. Then, the regenerated segment was studied histologically by haematoxylin-eosin staining and Van Gieson staining at 10 days, 3 weeks, 6 weeks, and 24 weeks postoperation. Retrograde urethrography was used to evaluate the function of the regenerated urethras of 4 rabbits 10 and 24 weeks after the operation. The urodynamics of 4 rabbits from the experimental group and control group I were assessed and compared. In addition, 4 experimental group rabbits were examined by a urethroscope 24 weeks after the operation.

RESULTSAt 10 days after operation, epithelial cells had migrated from each side, and small vessels were observed in the extracellular matrix. The matrix and adjacent areas of the host tissue were infiltrated with inflammatory cells. The epithelium covered the extracellular matrix fully at 3 weeks postoperation. Well-formed smooth-muscle cells were first confirmed after 6 weeks, at which point the inflammatory cells had disappeared. At 24 weeks postoperation, the regenerated tissue was equivalent to the normal urethra. Urethrography and urodynamic evaluations showed that there was no difference between normal tissue and regenerated tissue.

CONCLUSIONSUrethral extracellular matrix appears to be a useful material for urethral repair in rabbits. The matrix can be processed easily and has good characteristics for tissue handling and urethral function.

Animals ; Extracellular Matrix ; metabolism ; Rabbits ; Tissue Engineering ; methods ; Urethra ; pathology ; surgery

OBJECTIVETo estimate the effectiveness of inactivated influenza vaccine in elderly population.

METHODSAn quasi-experimental study was used. 590 elderly people who volunteered to receive the influenza vaccine were served as vaccine group, while 602 persons who did not want to receive the inoculation but could match the vaccine group were served as controls. One baseline and three follow-up surveys were carried out.

RESULTSThe protective rates of influenza like ill (ILI) as 52.38%, 36.84% and 37.89% with the decreasing rates of visits to ILI clinic as 45.16%, 50.54% and 50.54% were found after 1 month, 3 month and 6 month of inoculation of influenza vaccine; The protective rates of common cold, other respiratory tract or chronic disease were 49.54%, 64.54%, and 38.82%, respectively. The benefit-cost ratio was 4.98:1 in elderly population.

CONCLUSIONInfluenza vaccination could decrease ILI incidence and recurrence rates of related chronic diseases on elderly population to provide better economic benefits for the elderly.

Aged ; China ; epidemiology ; Cost-Benefit Analysis ; Female ; Humans ; Influenza Vaccines ; economics ; immunology ; Influenza, Human ; epidemiology ; prevention & control ; Male ; Middle Aged ; Vaccination

OBJECTIVETo express soluble HA of A/H1N1 influenza virus in drosophila S2 cell line and identify its bio-activity.

METHODSHA gene was amplified from A/Shenzhen/71/09 virus strain using RT-PCR, then we constructed pAC5.1-HA expression vector, which was co-transfected into S2 cell with pCoblast vector. After transfection, stable S2 cell was selected through Blasticindin. HA in the supernatant was identified with Western Blot assay and purified with Ni-column. Recombinant HA was immunized into BALB/c mice 3 times, and the Abs titers were evaluated with ELISA.

RESULTSWe successfully cloned HA gene with 1.7 x 10(3) bp of A/Shenzhen/71/09 virus strain and got recombinant pAC5. 1-HA expression vector. Stable S2 cell line was established after transfection and selection, which continuously expressed HA with molecular weight 75 x 10(3) D. After immunization with HA, the Abs titers were 1:1280 and 1: 5120 respectively on 10 d, 30 d.

CONCLUSIONWe expressed soluble HA with good bio-activity, which contributed to research on immune diagnosis, subunit vaccine, and monoclonal Abs for influenza.

Animals ; Blotting, Western ; Cell Line ; Drosophila ; Female ; Gene Expression ; Hemagglutinin Glycoproteins, Influenza Virus ; analysis ; chemistry ; genetics ; metabolism ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; metabolism ; Influenza, Human ; virology ; Mice ; Mice, Inbred BALB C ; Solubility

OBJECTIVETo construct full-length hepatitis B core particles presenting preS1 aa 21-47 epitope and truncated core particles presenting preS1 aa 37-45 epitope on their surface and compare their antigenicity.

METHODSPreS1 aa21-47 epitope and aa 37-45 epitope were inserted respectively into full-length hepatitis B core (aa 1-183) and truncated HBcAg (aa 1-144), between the 78th (Asp) and 79th (Pro). The genes synthesized after the codon optimization were ligated to the pET43. 1a vector with the same cohesive terminal (NdeI and XhoI) and expressed in the E. coli expression system. The morphology of the proteins of interest were observed by electron microscope and characterized by ELISA and Western Blotting.

RESULTSThe morphology of the virus-like particles were confirmed by electron microscope. H2 were solid particles with a diameter of (31.61 +/- 1.27) nm, while H3 were hollow particles with a diameter of (28.46 +/- 1.16) nm. Statistical analysis showed that H2 is larger than H3 in the diameter (P < 0.01). The antigenicity of the inserted epitopes and carrier protein were identified by ELISA and Western Blotting.

CONCLUSIONChimeric hepatitis B core particles presenting the preS1 neutralizing epitopes on their surface have been expressed, purified and identified, which lays the foundation for its application in vaccine research.

Epitopes ; chemistry ; genetics ; immunology ; Hepatitis B ; immunology ; virology ; Hepatitis B Core Antigens ; chemistry ; genetics ; immunology ; Hepatitis B Surface Antigens ; chemistry ; genetics ; immunology ; Hepatitis B virus ; chemistry ; genetics ; immunology ; Humans ; Neutralization Tests ; Protein Precursors ; chemistry ; genetics ; immunology ; Recombinant Fusion Proteins ; chemistry ; genetics ; immunology

OBJECTIVETo investigate the expression of translationally controlled tumor protein (TCTP) in human breast cancer tissues and its clinical significances.

METHODSThe expression of TCTP in 94 human breast cancer and the corresponding adjacent normal mammary tissues were detected using immunohistochemistry.

RESULTSThe expression rate of TCTP was 64.89% in human breast cancer tissues, significantly higher than that in normal benign mammary tissues (39.36%, P<0.001). TCTP overexpression was positively correlated to the tumor size, clinical stage, lymph node metastasis and histological grade of breast cancer (P<0.05). Patients with positive TCTP expression had a significantly shorter mean survival time than those with negative expression (P<0.001).

CONCLUSIONTCTP may play an important role in the tumorigenesis and development of breast cancer, and can be an important prognostic factor for this malignancy.

Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; Female ; Humans ; Middle Aged

OBJECTIVETo study the antigenic properties of mutant hepatitis B virus surface antigen, to understand the sensitivity of the commercially available HBsAg assays to the variants and to reduce the undetectability of the variants.

METHODSRecombinant eukaryotic expression plasmids for HBsAg. The recombinant eukaryotic expression plasmids pSS1adr, pSS1adw2, pSS1adw2- 145Arg, pSS1adr-126 Asn and pSS1adr-126Ser were transfected into COS-7 cells. HBsAg in the supernatants of transfected cells was detected by using different commercial ELISA kits.

RESULTSThe absorbance value of pSS1adr-126 Asn and pSS1adr-126Ser plasmids were similar to that of the wild type HBsAg, the absorbance value of pSS1adw2-145Arg plasmids was lower than that of the wild type HBsAg.

CONCLUSIONIt is estimated that the antigenicity of HBsAg mainly depended on the amino acid sequence of "a" antigen determinant and its conformation, so 145 amino acid substitutions led to the change of conformation and the antigenicity of variant HBsAg was lower than that of the wild type.

Animals ; COS Cells ; Cercopithecus aethiops ; Culture Media, Conditioned ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; genetics ; metabolism ; Hepatitis B Surface Antigens ; analysis ; genetics ; immunology ; Mutation ; Transfection ; Viral Envelope Proteins ; genetics

Objective To investigate the expression of glucagon-like peptide-1 receptor (GLP-1R) of skeletal muscle cells and the changes of skeletal muscle microstructure of diabetic rats, and to explore the relationship between the expressions of GLP-1R and insulin resistance in skeletal muscle. Methods Male Wistar rats were divided into two groups, normal glucose tolerance ( NT, n =25 ) , diabetes group ( DM group, n=30), the methods of creating diabetes model were used high-sugar high-fat diet and low-dose in-traperitoneal injection of streptozotocin. Immunohistochemical methods was used to detect the expression of GLP-1R in the muscle cells. The number of GLP-1R positive cells in the muscle of two groups was observed and compared. The relationship was analyzed between the homeostasis model assessment insulin resistance (HOMA-IR) and the number of GLP-1R positive cells. Results ⑴Compared to the NT group, the num-ber of GLP-1R positive cells in muscle of DM group was decreased (P<0. 05); ⑵ Compared to the NT group, HOMA-IR was increased in DM group (P<0. 05);⑶ Pearson correlation analysis showed that the percentage of GLP-1R positive cells expressed in rat skeletal muscle was negatively correlated with HOMA-IR (r= -0. 538, P<0. 01). Conclusions The expression of GLP-1R positive cells was decreased in the skeletal muscle of diabetic rats, insulin resistance increased. Therefore, the expression of GLP-1R on skele-tal muscle cells may promote the development of insulin resistance and diabetes mellitus.

Objective To investigate the effect of Zheng's 4C suspension transumbilical laparoendoscopic single-site surgery(TU-LSSS)in the treatment of cervical cancer,and analyze its prognosis.Methods A total of 92 patients with cervical cancer admitted to our hospital were selected and randomly divided into the control group(traditional laparoscopic surgery)and the observation group(Zheng's 4C suspension TU-LSSS),with 46 cases in each group.The perioperative related indicators of patients between the two groups were compared.The patients were followed up for 3 years,the progression-free survival(PFS)and overall survival(OS)were recorded,and the influencing factors of prognosis were analyzed.Results Compared with the control group,patients in the observation group had longer surgical time(P<0.05),less intraoperative bleeding(P<0.05),shorter recovery time for postoperative bowel sounds and hospital stay(P<0.05),higher postoperative 3-year PFS rate and 3-year OS rate(P<0.05).During 3-year follow-up,there were 11 cases of disease progression or death.The univariate analysis results showed that there were statistically significant differences in tumor diameter,clinical stage,lymph node metastasis,vascular infiltration,and treatment methods of patients between the good prognosis group and the poor prognosis group(P<0.05).The binary Logistic regression results showed that tumor diameter(≥4 cm),clinical stage(≥stage ⅠB2),lymph node metastasis,and vascular infiltration were the risk factors for the prognosis of patietns with cervical cancer(P<0.05),while Zheng's 4C suspension TU-LSSS was the protective factors(P<0.05).Conclusion Zheng's 4C suspension TU-LSSS can effectively promote the recovery of patients with cervical cancer,and improve 3-year PFS rate and 3-year OS rate.The prognosis is related to many factors,and targeted treatment should be performed according to different risk factors.

OBJECTIVETo evaluate the long-term clinical results and the factors that influences the outcomes of the revision open lumbar discectomy by fenestration.

METHODSFifty-one patients, who underwent the second open discectomy by fenestration from January 1 1988 to December 31 1994, were followed for an average of 146.8 months (range, 120 to 203 months). The long-term follow-up results were evaluated by using the MacNab classification and the Japanese Orthopaedic Association (JOA) scoring system through direct examinations and questionnaires. Radiography was also used in patients who agreed to visit the hospital, and findings were compared with those on preoperative radiographs.

RESULTSAt the final follow-up, with the MacNab classification an excellent and good outcome was achieved in 70.6% of the cases, 78.4% were satisfied with their results. The failure rate was 15.7% (8 patients). Excluding those 8 failed cases who needed another reoperation, the average improvement calculated by JOA scores was (64.6 +/- 18.2)%. The disc height of the operation site significantly decreased after surgery, nevertheless, this did not affect the long-term clinical outcome. Factors that were associated with a fair and bad outcome included smoking, isolated trauma or injury, fibrosis and the length of the remaining or recurrent primary postoperative symptoms history. Psychosociological signs were probably known as negative predictors of lumbar disc surgery outcome.

CONCLUSIONThe long-term outcome of the revision open lumbar discectomy by fenestration in this series was favorable. Because the revision operation is typically associated with a higher complexity, selection of suitable surgical candidates and determination of valid indications for operative treatment are very important. JOA scores have proved to be easy to perform for patients and clinicians and standardize subjective data.

Adult ; Aged ; Diskectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Reoperation ; Retrospective Studies ; Treatment Outcome