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

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.

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

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

Background: In China, most patients with nasopharyngeal carcinoma (NPC) are diagnosed at a late stage and con-sequently have a poor prognosis. This study aimed to investigate potential factors associated with the clinical stage of NPC at diagnosis. Methods: Data were obtained from 118 patients with early-stage NPC and 274 with late-stage NPC who were treated at Sun Yat-sen University Cancer Center between August 2014 and July 2015. Patients were individually matched by age, sex, and residence, and a conditional logistic regression model was applied to assess the associa-tions of clinical stage at diagnosis with socioeconomic status indicators, knowledge of NPC, physical examinations, patient interval, and risk factors for NPC. Results: Although knowledge of early NPC symptoms, smoking cessation, and patient interval were important fac-tors, the number of cigarettes smoked per day, motorbike ownership, and physical examination exhibited the strong-est associations with the clinical stage of NPC at diagnosis. Compared with smoking fewer than ten cigarettes a day, smoking 10–30 cigarettes [odds ratio (OR) 4.03; 95% confidence interval (CI) 1.11–14.68] or more than 30 cigarettes (OR 11.46; 95% CI 1.26–103.91) was associated with an increased risk of late diagnosis. Compared with not owning a motorbike, owning a motorbike (OR 0.38; 95% CI 0.23–0.64) was associated with early diagnosis. Subjects who under-went physical examinations were less likely to receive a late diagnosis than those who did not undergo examinations (OR 0.50; 95% CI 0.28–0.89). However, indicators of wealth were not significant factors. Conclusions: Initiatives to improve NPC patient prognosis should aim to promote knowledge about early symptoms and detection, health awareness, and accessibility to health facilities among all patients, regardless of socioeconomic status.

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

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 discuss the clinical findings and treatment of paraneoplastic pemphigus (PNP) with Castleman's disease.

METHODSTo investigate the clinical, histopathologic and CT findings of 8 cases paraneoplastic pemphigus with Castleman's disease.

RESULTSAll of 8 patients were diagnosed PNP first and were found Castleman's tumor incidently during routine examination. All 8 cases showed severe erosion or ulcer of the oral mucosa with various skin lesions. Histopathologically, there were intraepidermal acantholytic vesicle, basal cell liquefaction, necrotic keratinocytes in the epidermis and lymphocyte infiltration in the upper dermis. CT scan appeared solitary mass in these patients. Some of them were attacked by bronchiolitis obliterans. All 8 patients were failed by use of predisone. Obvious relief of PNP and pulmonary lesion occurred after tumor was rescted.

CONCLUSIONSParaneoplastic pemphigus with Castleman's disease is a rare disease. The key step is to find and resect the tumor in abdomen. CT scan should be used to detect the tumor in patients with PNP, especially, when predisone was failed in treatment.

Adolescent ; Adult ; Castleman Disease ; complications ; diagnosis ; therapy ; Combined Modality Therapy ; Female ; Humans ; Male ; Paraneoplastic Syndromes ; complications ; diagnosis ; therapy ; Pemphigus, Benign Familial ; complications ; diagnosis ; therapy ; Retrospective Studies