Objective To explore parkin gene deletion mutations at exons 3 to 7 in Chinese familial patients with Parkinson's disease as well as the association with the clinical features.Methods DNA was extracted from 6 unrelated families of PD patients; the deletion mutations of parkin gene at exons 3 to 7 were identified by PCR amplification, agarose gel electrophoresis. and the clinical data were analyzed together with the above information.Results In the 6 unrelated families of PD patients, 1 case had exon 5 deletion, its hereditary manner was autosomal recessive inheritance,the patient's age at the onset was 60 years old,clinical feature was tremor,rigidity and bradykinesia,but no athetosis.In addition the deletion mutations of parkin gene at exons 3,4,6,7 were not found.Conclusion There are deletion mutations of parkin gene at exons 5 in families PD of Chinese patients.

Objective To compare the characteristics of remifentanil and sufentanil for tracheal intubation general anesthesia in gyneco-logical laparoscopic operation. Methods 80 patients received gynecological laparoscopic operation were randomly divided into the remifen-tanil group and the sufentanil group with 40 patients in each group, and they were performed total intravenous anesthesia with remifentanil or sufentanil respectively. Compare the changes of heart rate, mean arterial pressure in the two groups before and after induction, at the time of intubation, pneumoperitoneum, and after the operation. And differences in two groups were recorded in the recovery time, extubation time, postoperative pain in 24 hours, exhaust time and postoperative complications. Results Change trend and magnitude in heart rate, mean ar-terial pressure had no significant difference in the two groups (P>0. 05). The recovery time, extubation time in remifentanil group was shor-ter (P<0. 01), postoperative pain in 24 hours in sufentanil group was more mild (P<0. 01). Exhaust time and postoperative complications of the two groups had no significant difference (P>0. 05). Conclusion Remifentanil and sufentanil used in gynecological laparoscopic op-eration can achieve good effect. and effects of the two drugs on hemodynamics were similar. Remifentanil can recieve better effect in terms of recovery time, while sufentanil has better postoperative analgesia effect.

Seeds of Panax quinquefolius L. with original moisture content of 5.9%, together withthat desiccated over silica gel to moisture contents of 3.4% and 1.7%, were stored at ambient tempera-ture,-5 ℃C, -18 ℃C and -196 ℃C (liquid nitrogen) for 6 months. Influences of desiccation and storage on seed viability were tested by TTC staining, before and after desiccation, and their vigour were evaluatedon tests of seed esterase isozyme isoelectric focusing (IEF), and field test on their rate of splitting and e-mergence,length of bud and root. Results showed that there was no effects on seed viability by desiccationto 3.4% and 1.7% of moisture content, as the TTC staining rates were 100% for all seeds tested immedi-ately after desiccation. No significant differences in seed viability and vigour were detected among seedsstored at defferent temperature as there are no significant difference in vairance analysis for seed split rateand seed emergency rate.

OBJECTIVETo produce anti-19-Nortestosterone (NT) monoclonal antibodies and identify their immunological characteristics.

METHODSHybridomas were prepared by fusing NS0 mouse myeloma cells with splenocytes isolated from immunized BALB/c mice. Noncompetitive and competitive indirect ELISA were employed to screen positive cell clones. A caprylic acid ammonium sulphate (CAAP) method was used to purify NT mAb, and the Batty saturation method was used to determine the affinity constant (Kaff).

RESULTSFive hybridoma cell lines, named NT-1, NT-2, NT-3, NT-4, and NT-5, were identified and their corresponding mAbs were of the IgG(1) isotype with a k light chain. The Kaffs of all mAbs were between 2.6 and 4.7 × 10(9) L/mol. The titers and IC(50) values of purified ascite fluids were in the range of (0.64-2.56) × 10(5) and (0.55-1.0) ng/mL, respectively. Of all the cross-reacting steroids, (-NT was the most reactive with the mAbs at 62% with NT-1 mAb and 64% with NT-2 mAb. Negligible cross-reactivity (<0.01%) with other steroids was observed.

CONCLUSIONThe establishment of these hybridomas allows the potential development of a rapid test kit, and may provide an alternative method for the detection of NT residues in food producing animals.

Animals ; Antibodies, Monoclonal ; Antibody Affinity ; Antibody Specificity ; Cell Line ; Female ; Hybridomas ; Mice ; Mice, Inbred BALB C ; Molecular Structure ; Nandrolone ; chemistry ; immunology ; Plasmacytoma ; Reagent Kits, Diagnostic ; Spleen ; cytology

Objective To identify the expression of CD68-tumor-associated macrophages (TAMs) and proliferative marker Ki-67 in retroperitoneal malignant fibrous histiocytoma (MFH) and their clinical significance. Methods Clinical data about 35 patients with retroperitoneal MFH managed with surgery from February 2002 to December 2015 were retrospectively analyzed and all patients were followed up. There were 24 male and 11 female patients, and they were 18-71 years old, with mean age (53.0 ± 10.8) years old. Patients were divided into CD68 positive group (21 patients) and CD68 negative group (14 patients), while they were also divided into Ki-67 low expression (< 20%) group and Ki-67 high expression ( ≥ 20%) group, according to the immunohistochemical staining results. The overall survival time and all clinical data between two groups were compared. Kaplan-Meier estimations, Cox regression analysis, Fisher exact probabilities and Spearman correlations were performed. Results Of the 35 patients, 18 patients received radical resection, and 17 patients received palliative operation. The overall 1-, 3-, 5-year survival rates were 65.7%, 22.9%and 8.6%and the median survival was 17 (1-86) months. Factors associated with postoperative survival were FNCLCC grade (x2=7.002, P=0.008), modusoperandi of the tumor resection(x2=7.134, P=0.008), and CD68(x2=4.634, P=0.031) and Ki-67 overexpression (≥20%) (x2=8.898, P=0.003 ) . The difference between gender, age, tumor size, blood loss, removal of the joint organs and adjuvant therapy got no statistical significances (P > 0.05). Multivariate analysis showed that survival was associated with modusoperandi of the tumor resection and Ki-67 overexpression (P=0.003, 0.002, respectively). Conclusions Retroperitoneal malignant fibrous histiocytoma is a rare malignancy that display poor prognosis and high mortality. Complete resection remains the mainstream for retroperitoneal malignant fibrous histiocytoma. The patients' life span in CD68 positive or Ki-67 high expression is shorter. CD68 and Ki-67 plays a critical role in retroperitoneal malignant fibrous histiocytoma carcinogenesis and their high expression may be used as a potential survival predictor in patients with retroperitoneal MFH.

Objective To investigate a convenient and quantitative solution to activation levels and functional characterization of CAR-T cells by inserting T cell activity-responsive promoter (TARP) nanoluciferase reporter gene system into a lentiviral plasmid containing the gene encoding the chimeric antigen receptor (CAR). Methods The recombinant plasmid was constructed by using whole gene synthesis and molecular cloning techniques. The lentivirus was packaged and was infected with human primary T lymphocytes. Flow cytometry was used to detected the positive rate of lentivirus-infected T cells. The functional characterization of CAR-T cells was identified by luciferase reporter gene system, Western blot, flow cytometry, and small animal live imaging techniques. Results The results of enzyme digestion identification and the plasmid sequencing showed that the recombinant plasmids were constructed, and flow cytometry displayed the normal preparation of CAR-T cells. This system could dynamically respond to the activation of CAR-T cells by luciferase reporter gene system. The functional assay in vitro confirmed that the system could reflect the exhaustion of CAR-T cells, and the small animal live imaging results demonstrated that the system can be used as a tracer of CAR-T cells in mice. Conclusion TARP nanoluciferase reporter gene system provides a more convenient, sensitive and quantitative method for evaluating CAR-T cells activation level, exhaustion phenotype and tracing.
Humans ; Animals ; Mice ; T-Lymphocytes ; Cell Line, Tumor ; Receptors, Chimeric Antigen/genetics* ; Promoter Regions, Genetic ; Immunotherapy, Adoptive/methods*