Objective To construct the eukaryotic expression vector of linker for activation of T cells(LAT) gene in mouse. MethodsLAT cDNA of mouse mononuclear cells was amplified with polymerase chain reaction.The fragment was inserted into the eukaryotic expression vector of pCMV-HA plasmid.The recombinant plasmid was verified by DNA sequencing and used to transfect lymphocytes of the asthmatic model. Results The length of amplified fragment was 729 bp.The sequence of LAT gene was confirmed by Genbank.The LAT protein could be detected in the lymphocytes of asthmatic model.Conclusion The recombinant eukaryotic expression vector pCMV-HA-LAT can be successfully constructed.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast ; pathology ; Breast Neoplasms ; drug therapy ; pathology ; Cyclophosphamide ; therapeutic use ; Diagnosis, Differential ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Lymphoma, B-Cell ; drug therapy ; pathology ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; pathology ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Sarcoma ; pathology ; Spleen ; pathology ; Splenic Neoplasms ; drug therapy ; pathology ; Vincristine ; therapeutic use

OBJECTIVETo investigate the effectiveness of surgical strategies for Shang Ring circumcision in the treatment of short frenulum praeputii in patients with redundant prepuce or phimosis.

METHODSTotally, 130 cases of short frenulum praeputii with redundant prepuce or phimosis were randomly assigned to an experimental group and a control group of equal number to receive Shang Ring circumcision, the former by transverse incision in the distal penis foreskin and pull-up of the interior board, and the latter by conventional transverse incision and longitudinal suture of the frenulum praeputii. Comparisons were made between the two groups in the surgical duration, intraoperative blood loss, 24 h postoperative pain visual analog score (VAS), postoperative complications, satisfaction with the penile appearance, and the quality of sexual life.

RESULTSThe surgical duration, intraoperative blood loss, 24 h postoperative VAS, postoperative sexual satisfaction, and satisfaction with penile appearance were (4.60 +/- 1.20) min, (2.61 +/- 1.81) ml, 1.73 +/- 0.76, 98.5%, and 98.5%, respectively, in the experimental group, as compared with (21.60 +/- 6.30) min, (11.10 +/- 3.40) ml, 5.37 +/- 1.84, 70.3% and 69.8% in the control, with statistically significant differences between the two groups (P < 0.05). The incidence rates of such major complications as wound dehiscence, infection, and moderate to severe edema were 1.5% (1/65), 3.1% (2/65), and 4.6% (3/65), respectively, in the experimental group in comparison with 12.3% (8/65), 15.3% (10/65), and 30.7% (20/65) in the control, with statistically significant differences between the two groups (P < 0.05). None of patients had any serious complications.

CONCLUSIONShang Ring circumcision by transverse incision in the distal penis foreskin and pull-up of the interior board, with its advantages of shorter operation time, less blood loss, mild pain, fewer complications, and higher satisfaction and acceptance of the patients, can be used as an safe and effective approach to the treatment of short frenulum praeputii.

Aged ; Blood Loss, Surgical ; statistics & numerical data ; Circumcision, Male ; adverse effects ; instrumentation ; methods ; Edema ; epidemiology ; Foreskin ; abnormalities ; surgery ; Humans ; Incidence ; Male ; Operative Time ; Pain Measurement ; Pain, Postoperative ; diagnosis ; Patient Satisfaction ; Phimosis ; surgery ; Postoperative Period ; Prostheses and Implants ; Surgical Wound Dehiscence ; epidemiology ; Surgical Wound Infection ; epidemiology

Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software.Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template.The amplified fragments G1 and G2 are 570bp and 308bp in length,respectively.The PCR products were cloned into pET30a vector and expressed in soluble form in E.coli after induction of cultured E.coli with IPTG.Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions.Then the purified proteins were analysed by Western blotting.The results showed that the purified recombinant protein G1 retained good antigenicity and specificity.But the purified recombinant protein G2 didn't possess biological activity.Antibodies against BRSV were detected in suspected bovine serum samples in China by using indirect ELISA and Western blotting with the purified recombinant protein G1.The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection.And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.

Abstract: Objective　To analyze the distribution and drug resistance evolution characteristics of pathogenic bacteria of bloodstream infection in nine tertiary hospitals in Yunnan Province from 2017 to 2021, so as to provide reliable basis for rational selection of antibiotics in clinic. Methods Using the drug sensitive paper method or instrument method, the bacteria identification and drug sensitivity test were carried out in nine tertiary hospitals in different regions according to the unified technical scheme. The results were judged according to the Clinical and Laboratory Standards Institute (CLSI) breakpoint standard in 2021, and use WHONET5.6 for data statistical analysis. Results A total of 12 003 strains of pathogenic bacteria were isolated from bloodstream infection samples in the past five years, including 7 442 strains of Gram-negative bacteria (62.0%) and 4562 strains of Gram-positive bacteria (38.0%), with an increasing trend in the number of isolated strains; of these, 163 strains (1.4%) were isolated from outpatients and 11 840 strains (98.6%) were isolated from inpatients. The top three gram-negative bacteria were Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii, of which 309 strains (4.2%) were carbapenem-resistant Klebsiella pneumoniae (CR-KPN), 29 strains (0.4%) carbapenem-resistant Escherichia coli and 19 strains (0.3%) carbapenem-resistant Enterobacter cloacae, and the number of CR-KPN was on the rise year by year. The top three Gram-positive bacteria were coagulase-negative staphylococci, Staphylococcus aureus and Enterococcus faecium, of which methicillin-resistant Staphylococcus aureus (MRSA) was detected for 213 strains, accounting for 27.7%, and decreased from 40.0% in 2017 to 23.4% in 2021, showing a downward trend year by year. No vancomycin-resistant staphylococci and enterococci were found. Conclusions The detection and composition of bloodstream infection pathogenic bacteria in multicenter have not changed much in the past five years, but each hospital has its own characteristics. The number of carbapenem resistant Enterobacteriaceae increased year by year, which should be paid more attention.

Solid dispersions technique was used to solidify buagafuran and improve buagafuran in vitro dissolution and stability. Buagafuran solid dispersions were prepared separately using PVPK30, PEG6000 and Poloxamer188 at various weight ratios as carriers. The status of buagafuran in solid dispersions was determined by using DSC and IR. The solubility, content and in vitro dissolution of pure drug and the solid dispersions were detected by using HPLC. When buagafuran/carrier was 1:5 or less, the drug existed in a solid dispersion form. Three kinds of carriers all can improve buagafuran dispersibility and in vitro dissolution. Accelerating experiment showed that buagafuran/PVPK30 < or = 1:10 solid dispersions was ageing-resistant, and the aspect, content and in vitro dissolution did not change after storaged over 3 months, but PEG6000, Poloxamer188 and a lower ratio PVPK30 solid dispersions became aged. Buagafuran/PVPK30 < or = 1:10 solid dispersions can be developed as buagafuran oral drug delivery carrier.
Anti-Anxiety Agents ; administration & dosage ; chemistry ; Drug Carriers ; Drug Delivery Systems ; Drug Stability ; Drug Storage ; Poloxamer ; chemistry ; Polyethylene Glycols ; chemistry ; Povidone ; chemistry ; Powders ; Sesquiterpenes ; administration & dosage ; chemistry ; Solubility

OBJECTIVETo study the ability of bone marrow mesenchymal stem cells (BMSCs) to repair the internal environment of the testis in male azoospermia rats.

METHODSWe established azoospermia models in 22 six-week-old male SD rats by intraperitoneal injection of busulfan at 20 mg per kg body weight. We transplanted allogeneic rat BMSCs (rBMSCs) into the testicular seminiferous tubules of the model rats and, 30 days after transplantation, observed the composition and structure of the seminiferous tubular cells by HE staining and detected the expressions of CD44, CD106, and c-kit in the rBMSCs by immunohistochemistry.

RESULTSThe number of epididymal sperm was significantly reduced in the model rats as compared with the normal controls (P < 0.01). CD44 and CD106, but not c-kit, were expressed in the isolated rBMSCs. At 30 days after transplantation of rBMSCs, lots of new cells were observed in the seminiferous tubules, some expressing CD106 and some expressing the germ cell surface marker c-kit.

CONCLUSIONBMSCs can transdifferentiate into germ cells and repair the damaged seminiferous tubules of sterile rats.

Animals ; Azoospermia ; chemically induced ; therapy ; Biomarkers ; metabolism ; Bone Marrow Cells ; Busulfan ; Cell Membrane ; metabolism ; Epididymis ; Hyaluronan Receptors ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Proto-Oncogene Proteins c-kit ; metabolism ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; anatomy & histology ; metabolism ; Spermatozoa ; Staining and Labeling ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo evaluate the expression of galectin-3 and galectin-9 in mice nasal mucosa,and to explore the role of galectin-3 and galectin-9 in allergic rhinitis (AR).

METHODSTwenty mice were randomly divided into AR group and control group, 10 mice in each group. Ten mice of BALB/c were sensitized intraperitoneally with 10 microg of ovalbumin(OVA) adsorbed onto 2 mg Al(OH)3 on day 1 and day 14. Mice was induced daily by intranasal daily administration of 10 microl of saline containing 100 microg of OVA from day 21 to day 28. OVA was replaced with saline in control group. Immunohistochemical staining was used to examine the expression of galectin-3 and galectin-9 in nasal mucosa. RT-PCR was performed to investigate the level of mRNA expression of complement galectin-3, galectin-9. The level of IL-4, IL-5 and IFN-gamma in peripheral blood were titrated by ELISA.

RESULTSGalectin-3 and galectin-9 were detected in both groups. Expression of galectin-3 and galectin-9 in group AR was higher than that in control group. Pearson correlation analysis demonstrated that the levels of galectin-3 and galectin-9 were positively correlated with the level of IL-4 and IL-5, but negatively correlated with the level of IFN-gamma.

CONCLUSIONSGalectin-3 and galectin-9 might play an important role in the pathogenesis of allergic rhinitis.

Animals ; Female ; Galectin 3 ; metabolism ; Galectins ; metabolism ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Interleukin-5 ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Seasonal ; metabolism

AIMTo observe the influence of polysaccharides of Angelica sinensis (ASP) on the immunologic function of rat Kupffer cells.

METHODSNormal rat Kupffer cells were treated with ASP in vitro. Absorbance at 540 nm ( A540) of neutral red absorption and supernatant NO, TNF-alpha in the cells were measured to evaluate the immunologic function of Kupffer cells; LDH leakage was measured to estimate the severity of cellular damage; Rats were given ASP 0.025, 0.1, 0.25 and 1.0 g x kg(-1) ig (qd x 7 d) in vivo. The above indices and ACP of Kupffer cells were measured, sGST and sALT activity were detected as indices of hepatotoxicity.

RESULTSASP markedly enhanced the phagocytic activity, ACP and supernatant NO, TNF-alpha of Kupffer cells both in vitro and in vivo . The increase of sGST was observed after administration of ASP 1.0 g x kg(-1), but the LDH leakage of the hepatocytes was not increased in vitro.

CONCLUSIONASP with suitable dose could activate the function of Kupffer cells. Slight liver injury was caused by ASP 1.0 g x kg(-1) in vivo, which was likely caused by factors, such as NO, TNF-alpha, indirectly.

Adjuvants, Immunologic ; pharmacology ; Alanine Transaminase ; blood ; Angelica sinensis ; chemistry ; Animals ; Cells, Cultured ; Female ; Glutathione Transferase ; blood ; Hepatocytes ; metabolism ; Kupffer Cells ; immunology ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Male ; Nitric Oxide ; metabolism ; Phagocytosis ; drug effects ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo analyze the electrophysiological characteristics and efficacy of radiofrequency catheter ablation (RFA) of focal atrial tachycardia (AT) originating from the left atrial appendage (LAA).

METHODSElectrophysiologic study and RFA were performed in 9 patients (4 female) with focal AT originating from the LAA. Atrial appendage angiography was performed to identify the origin of AT. P waves were classified as negative, positive, isoelectric, or biphasic.

RESULTSThe mean age was (21 +/- 9) years. AT occurred spontaneously or was induced by isoproterenol infusion rather than programmed extrastimulation and burst atrial pacing. A characteristic P-wave morphology and endocardial activation pattern were observed. Positive P-wave in inferior leads was seen in all patients, upright or biphasic (+/-) component P wave was observed in lead V1, isoelectric component or an upright component P wave with low amplitude ( < 0.1 mV) was seen in lead V2-V6. Earliest endocardial activity occurred at the distal coronary sinus (CS) in all patients. The earliest endocardial activation at the successful RFA site occurred (36.7 +/- 7.9) ms before the onset of P wave. RFA was successful in all 9 patients immediately post procedure. AT reoccurred in 2 patients within 1 month post RFA and AT disappeared post the 2nd-RFA. AT reoccurred in 1 patient and terminated after the 3rd RFA. At the final follow-up (12 +/ 5) months, all 9 patients were free of arrhythmias without antiarrhythmic drugs.

CONCLUSIONSThe LAA is an uncommon site of origin for focal AT. The characteristic P wave and activation timing are suggestive for focal AT originating from the LAA. LAA focal ablation is safe and effective for patients with focal AT originating from the LAA.

Adolescent ; Adult ; Atrial Appendage ; physiopathology ; Catheter Ablation ; methods ; Child ; Electrophysiological Phenomena ; Female ; Humans ; Male ; Tachycardia, Ectopic Atrial ; physiopathology ; surgery ; Treatment Outcome ; Young Adult