Objective To prepare monoclonal antibody(McAb) against human tissue kallikrein (HK) and develop an ELISA kit allows for the in vitro quantitative determination of human tissue kallikrein in urine. Methods To generate a monoclonal antibody specific for TK, the synthetic TK peptide consisting of 12 amine acids(12P), was fused to keyhole limpet hemocyanin(KLH) and used for immunization. Using hybridoma screening, monoclonal secreting cell lines were identified and used to generate ascites in BALB/c mouse. Antibody was purified by affinity column chromatography. 12% SDS-PAGE and Western blot were used to visualize the purified antibody. This kit employs indirect competitive ELISA technique and BiotinAvidin System. 12P was fused to bovine serum albumin(BSA) and has been pre-coated onto a microplate at first. Standards and samples were added to the appropriate microplate wells with a biotin-conjugated McAb croplate well. A TMB substrate solution is added to each well. The enzyme-substrate reaction is terminating by the addition of a sulphuric acid solution and the color change is measured spectrotometrically at a wavelength of 450 nm. The concentration of tissue kallikrein in the samples is then determined by comparing the O.D. of the samples to the standard curve. Results 8 hybridoma cell lines secreting mAbs special to HK,SDS-PAGE and Western blot demonstrated successful preparing and purification of McAb( 100% ). The linearity of this ELISA kit is demonstrated(r =0. 990). The range of detection of the assay is 0.008 μg/ml to 0. 5 μg/ml. The assay remained stable, with no change in the values measured, over five cycles of freezing and thawing. Conclusion 8 McAbs against HK have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for detecting HK can meet the needs of detection of HK in urine samples.

ObjectiveTo evaluate the assistant effct of electroacupuncture on acute incomplete spinal cord injury treated with routine therapy.Methods9 cases who accepted routine therapy, including operation and medicine, were in control group, while other 7 cases who accepted electroacupuncture other than routine therapy were in treating group.ResultsThere were 2 cases were effective, 4 cases were improved and 3 cases were not improved in control group, while 6 cases were clinical recovery, 1 case was effective in treating group.ConclusionElectroacupuncture can improve the effect of routine therapy on acute incomplete spinal cord injury.

目的观察神经外科与针刺联合治疗急性脊髓不完全损伤的疗效。方法神经外科治疗组根据指征确定是否进行手术,药物治疗均给予激素、脱水、抗去甲肾上腺素类药物、神经营养药物,高压氧。神经外科与针刺联合治疗组除上述神经外科治疗外,采用针刺治疗,针刺处方:电针刺激夹脊穴,根据不同辨证配合远端取穴。结果神经外科治疗组9例,持续治疗 3—6个月,出院后随访6个月,6例好转,肌力有所改善,最好者可达到肌力IV—IV+级,3例无效,甚至退步,瘫痪进一步加重。神经外科与针刺联合治疗组病人5例,经针刺及电针治疗10—80天时间后,4例临床治愈,1例经针刺治疗好转后,因转入骨科手术治疗腰椎压缩性骨折时脱离针刺治疗。结论神经外科与针刺联合治疗急性脊髓损伤效果好,并能抑制脊髓损伤的进一步恶化。

Objective To establish a stable and efficient method of culturing imDCs in vitro,and to explore the effect of GW5074, which blocks ERK1/2 signal pathway in the process of imnature dentritic cells (imDCs) on inducing differentiation of the na(i)ve allogeneic CD4+ T cells into Treg cells in vitro. Methods The imDCs and mature DCs (mDCs) were isolated and cultured from the peripheral blood mononuclear cells (PBMC) derived from a healthy adult male volunteer, and they were identified by cell morphology, cell surface marker and cell functions respectively. Na(i)ve CD4+ T cells were isolated from newborn umbilical vein blood and were divided into 5 groups to be cultured: (1) Blank control group: Na(i)ve CD4+ T cells were cultured alone;(2) Positive control group: The irrDCs were Middle-concentration GW5074 group;(5) High-concentration GW5074 group. In the last three groups, imDCs and na(i)ve CD4+ T cells were co-cultured, the same as the positive control group, but these groups were added by GW5074 dilution at the concentrations of 8, 24, and 40μmol/Lrespectively. After co-culture for 5 days, the transformation ratio from naive CD4+T cells to Treg T cells was detected by flow cytometry. Results On the surface of imDCs, there was stronger pression of CD1a, but weaker expression of CD80 and CD83. On the contrary, on the surface of mDCs, there was weaker expression of CD1a, but stronger expression of CD80 and CD83. The stimulation index in imDCs group and mDCs group was 1.12±0.03 and 2.85±0. 07 respectively. The transformation ratio of Treg T cells in blank control group, positive control group, low-concentration GW5074 group, middle-concentration GW5074 group and high-concentration GW5074 group was (5. 81±1.36)%, (35.73±2.07)%, (22.53±2.11)%, (11.55±1.73)%, and (4.97±1.83)%respectively. One-way ANOVA analysis revealed that there was no significant difference between high-concentration GW5074 group and blank control group, P＞0. 05, but significant difference between the remaining groups, P＜0.01. Conclusion High purity of imDCs can be obtained from PBMC by induction with rhGM-CSF and rhIL-4. ERK1/2 signal pathway plays a role in inducing the immune tolerance. GW5074 can inhibit differentiation of na(i)ve CD4+ T cells into Treg T cells.

OBJECTIVETo explore the expression of MMP2 and its correlation with the clinical features and prognosis of endometrial adenocarcinoma.

METHODSWe collected paraffin-embedded samples from 81 patients with endometrial adenocarcinoma aged 32 to 80 years to examine the expression of MMP2 using immunohistochemistry. The correlation of MMP2 expression with the clinical characteristics and prognosis of the patients was analyzed.

RESULTSMMP2 protein was expressed in the cytoplasm of the tumor cells. MMP2 over-expression was negatively correlated with tumor differentiation (P=0.015) and prognosis (P=0.041) of endometrial adenocarcinoma.

CONCLUSIONMMP2 over-expression is a potential malignant biomarker in endometrial adenocarcinoma.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Endometrial Neoplasms ; metabolism ; pathology ; Female ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Middle Aged

OBJECTIVE To analyze the current status and research hot spots of traditional Chinese medicine （TCM）in the prevention and treatment of myocardial ischemia -reperfusion injury （MIRI），and to provide reference for developing relevant studies. METHODS A bibliometric analysis was performed with the literature on TCM treatments for MIRI included in the Web of Science as the data source . RESULTS A total of 972 literatures were included in this study . The research on TCM for prevention and treatment of MIRI in recent 10 years showed an increasing trend year by year ，and the literature published in 2021 were cited most frequently ；most research scholars focused in China ；Peking Union Medical College ，Chinese Academy of Medical Sciences had the most papers in this field ，and a lot of collaboration with other institutions ，which had a large influence in the field ；Sun Guibo and Sun Xiaobo were high -yielding authors in the field ，but the global authorship was not very close and had not formed cooperative teams of some scale ；Molecular Medicine Reports was perhaps the most popular journal in the field . Co-citation analysis found that the research direction in this field was mostly focused on exploring the pathogenesis of MIRI and finding intervention targets . Key words analysis revealed that TCM regulated apoptosis ， pyroptosis， mitochondrial dysfunction ， miRNA and other pathways to improve MIRI research was a hot spot in the field ，and more studies had focused on the Nrf 2 pathway，NF-κB pathway ，NLRP3 inflammasomepathway， TNF pathway ， SIRT3 pathway， JNK pathway ，PI3K/Akt pathway and AMPK pathway . CONCLUSIONS Much attention has been paid to the prevention and treatment of MIRI by TCM in the past 10 years，and the current hot spots in this field are mostly focused on the studies about the intervention mechanism of TCM ’s active ingredients and Chinese patent medicines on MIRI . Moreover，the research about TCM in the treatment of MIRI is mostly focused on the field of basic experiments at present ，and it is recommended that many large -scale，high-quality， multicenter，randomized controlled clinical studies should be carried out in the future ，to provide high -grade evidence -based evidence for the application of TCM in clinic .