Stroke is one of the main causes of death in the world, of which more than 85% are acute ischemic stroke, and reperfusion therapy is its main treatment. Studies have shown that the neutrophil to lymphocyte ratio (NLR) is associated with the efficacy, safety and outcome of the reperfusion therapy. This article reviews the predictive role of NLR in patients with ischemic stroke receiving reperfusion therapy.

Objective To investigate the relationship between plasma soluble CD36 (sCD36) and nonalcoholic fatty liver disease (NAFLD) in patients combined with type 2 diabetes mellitus (T2DM). Methods Plasma levels of sCD36 were determined in normal control group (group A, n=39), patients of T2DM without NAFLD group (group B, n=39) and T2DM with NAFLD group (group C, n=59). Liver fat content (LFC) and nonalcoholic fatty liver fibrosis score (NFS) were calculated in group C. Glucose and lipid metabolic parameters, liver function parameters and inflammatory parameters were also detect?ed in all three groups. Variance analysis was applied to analyze the differences of the above parameters among three groups;Correlation analysis was used to analyze the relationship between sCD36 level and all the above parameters;Multiple step? wise regression analysis was applied to determine the influencing factors of sCD36 level in patients of group C. Results Plasma sCD36 (μg/L) levels in group B (3.87 ± 1.16) and group C (5.72 ± 1.79) are higher than that of group A (2.57 ± 0.93) (both P<0.01), and it is higher in group C than in group B (each P<0.05);Correlation analysis showed that sCD36 level was positively correlated with body mass index (BMI), waist, visceral adipose tissue,fast insulin (FINS), insulin resistance in?dex (HOMA-IR), free fatty acid (FFA), alanine transaminase (ALT), tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6), LFC and NFS (P<0.01 or P<0.05);Multiple stepwise regression analysis showed that FFA, LFC, TNF-αand IL-6 were in?fluencing factors of sCD36 level in patients of group C. Conclusion Plasma sCD36 level was related to fatty liver severity, liver injury and fatty liver fibrosis, it might be used as a plasma marker of T2DM combined with NAFLD. CD36 might con?tribute to the development of T2DM combined with NAFLD through inflammatory mechanisms.

Objective To study the relationship between advanced liver fibrosis and peripheral neuropathy in patients with type 2 diabetes mellitus (DPN). Methods A total of 173 patients (89 men and 84 women) with type 2 diabetes who hos?pitalized in Tianjin Third Central Hospital within nearly three years (2013.02-2015.02) were divided into three groups ac?cording to non-alcoholic fatty liver disease (NAFLD) fibrosis score:group A (NFS≤-1.455), group B (-1.455

Objective To construct eukaryotic expressing vector of mouse soluble CD160 and stably transfect into CHO cells for eukaryotic expression.Methods Recombinant soluble CD160(rsCD160) was constructed by gene recombination.Total RNA was extracted from the spleen of C57BL/6 mice.cDNA was amplified for the soluble form of CD160.Then,the PCR product was cloned tO pcDNA3.1 and pEGFP-N1.The recombinant plasmid was identified by restriction map and sequence analy-sis.The soluble CDl 60 expression in CHO cells transfected with recombinant psCDl 60 was verified by RT-PCR and Westernblot.The binding ability of psCD160 tO its ligand was detected by FACS.Results 520 bp mouse soluble CD160 gene was obtained.Recombinant mouse psCD160 was successfully constructed.After transfection,soluble CD160 expression in the culture supernatant of CHO cells was successfully detected.FACS analysis indicated that soluble CD160 could bind tO its ligand.Conclusion Recombinant mouse psCD160 is successfully constructed,which will benefit our further study on soluble CD160 for immune therapy against tumor in the future experiments.

Objective:To investigate the risk factors and treatment outcome of recurrent Acanthamoeba keratitis (AK) after corneal transplantation. Methods:A serial case-observational study was carried out.Twenty-eight eyes of 28 patients with AK who underwent corneal transplantation in Shandong Eye Hospital from January 2012 to January 2019 were enrolled.All the eyes received corneal transplantation from failing to respond to topical and systemic anti- Acanthamoeba medical therapy, including 13 eyes that received penetrating keratoplasty (PKP) and 15 eyes that received lamellar keratoplasty (LKP). The corneal lesion was removed by a trephine with a diameter of 0.5 mm over infiltration area during PKP or LKP.The clinical features of recurrent AK were summarized, including recurrence time, site and signs, and the risk factors of AK recurrence were analyzed.Local and systemic anti- Acanthamoeba medical therapy was performed in all relapsed eyes, and secondary surgery was performed for the eyes with poor response to medication.The therapeutic outcome of recurrent AK was evaluated.The study adhered to the Declaration of Helsinki.This study protocol was approved by an Ethics Committee of Shandong Eye Hospital (No.201112). Results:In the 28 eyes, 7 eyes (25%) appeared recurrent AK after keratoplasty, including 2 eyes after PKP and 5 eyes after LKP.There was no significant difference in the recurrence rate between the two methods ( P=0.396). The recurrence rate of eyes that had used glucocorticoids drugs before operation was 57.14% (4/7), which was significantly higher in comparison with 14.29% (3/21) of eyes without glucocorticoids before surgery ( P=0.043). The recurrence rate of eyes with ulcer diameter ≥8.2 mm was 50.00% (5/10), which was significantly higher than 11.11% (2/18) of eyes with ulcer <8.2 mm ( P=0.036). The recurrent lesions began at the edge of implant bed accounted for 85.71% (6/7), and the recurrent lesions located below graft accounted for 14.29% (1/7). In 7 eyes with recurrent AK, 6 eyes were completely cured.Among recurrent AK eyes after LKP, 2 eyes were cured by long-term medical therapy, and 2 eyes were cured by extended-diameter LKP, and another 1 eye was cured by conjunctival flap covering surgery.One eye with recurrent AK after PKP was cured by extended-diameter PKP. Conclusions:The risk factors of recurrent AK after surgery are application of glucocorticoids before surgery and big lesions.Recurrent AK after surgery is curable by individualized therapy targeting to different clinical characteristics.

OBJECTIVETo investigate the effect of stable knockdown of DNA methyltransferase 3b (DNMT3b) on the proliferation and apoptosis of bladder cancer cells.

METHODSLentivirus expressing DNMT3b siRNA or the negative control siRNA was infected in human bladder cancer BIU-87 cells. MTT assay and flow cytometry were used to detect cell proliferation and apoptosis, respectively. The inhibitory effect of DNMT3b knockdown on xenograft tumors in nude mice was observed. Real-time PCR and Western blotting were carried out to investigate the expression level of cell apoptosis related genes. Methylation specific PCR was used to examine the methylation in the promoter region of the cell apoptosis related genes.

RESULTSThe results of real-time PCR and Western blotting showed that DNMT3b mRNA and protein level were stably knocked down in BIU-87 cells. Stable DNMT3b knockdown suppressed BIU-87 cell growth and the tumor formation ability of the cells in nude mice. DNMT3b knockdown promoted the apoptosis of BIU-87 cells, increased the mRNA and protein expression of the cell growth and apoptosis related genes including DAPK, Bax and RASSF1A, and significantly decreased the methylation of these genes.

CONCLUSIONStable DNMT3b knockdown can affect the methylation of the cell growth and apoptosis related genes to regulate their expression, which might be a possible mechanism for suppressed cell growth and enhanced apoptosis of BIU-87 cells.

Animals ; Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; Gene Knockdown Techniques ; Humans ; Mice ; Mice, Nude ; Neoplasm Transplantation ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Urinary Bladder Neoplasms ; genetics ; pathology

Objective:To construct the evaluation index system of lumbar puncture teaching for medical students, aiming to create a scientific assessment and evaluation method covering theoretical knowledge, skill practice, and professional accomplishment, so as to comprehensively evaluate the teaching effect of lumbar puncture for medical students, and improve the practical ability of clinical skills of medical students more effectively.Methods:The evaluation index scheme of lumbar puncture teaching for medical students was initially formulated through literature review and group discussion, and 20 experts related to clinical front-line work and medicine were invited to revise the scheme by applying Delphi expert consultation and applying analytic hierarchy process to quantify the entries and establish the final index weights at all levels.Results:The valid recovery rate of both rounds of expert consultation questionnaires in this study was 100%. The second round of expert consultation was conducted without changing experts, with an authority factor of 0.88. Kendall's coefficient of harmony was 0.136 and 0.184, respectively. After two rounds of expert consultation, the evaluation index system of lumbar puncture teaching for medical students, including 3 primary indicators (theoretical knowledge, comprehensive clinical ability and professionalism), 7 secondary indicators and 22 tertiary indicators, was initially constructed.Conclusion:The evaluation index system of lumbar puncture teaching for medical students constructed in this study is scientific and credible, which can provide quantitative basis for the training and assessment of medical students, and is of great theoretical and practical significance.

【Objective】 To investigate the effect of autologous blood transfusion(ABT) drainage system and simple drainage(using drainage bags) on the prognosis of patients after lumbar surgery. 【Methods】 The patients admitted to the Department of Orthopedics of our hospital from August 2018 to September 2020 who underwent posterior open lumbar internal fixation and fusion were divided into two groups according to different drainage methods adopted after surgery: 50 patients were randomly selected from the patients who received postoperative ABT system for drainage as ABT group, and 50 patients were randomly selected from the patients who received postoperative drainage by drainage bag as simple drainage bgroup(the control group). The postoperative drainage volume, actual postoperative drainage, total dominant blood loss, total autologous blood transfusion volume, as well as the postoperative anemia indexes, infection indexes and albumin levels in d1, d3 and d7 of the 2 groups were retrospectively analyzed. 【Results】 The gender, age, operation duration and operation segment of the 2 groups were comparable (P>0.05), and preoperative Hb, Hct, ALB, WBC, NE%, intraoperative blood loss, intraoperative autologous blood transfusion volume and actual postoperative drainage volume were similar (P>0.05). There were no significant differences in Hb, Hct, ALB, WBC, NE% and CRP in postoperative d1, d3 and d7, as well as in preoperative and postoperative Hb, Hct and ALB, and in postoperative and preoperative WBC, NE% and CRP (P>0.05). The postoperative drainage volume (mL) and total dominant blood loss (mL) in ABT group and the control group were 554.40±176.82 vs 337.80±102.43, and 1 048.40±282.87 vs 791.80±277.02, respectively (P<0.05). 【Conclusion】 The use of ABT drainage system after lumbar surgery increased the drainage volume compared with simple drainage, but the improvement of anemia, albumin and infection was not obvious. ABT system should not be used routinely for drainage after lumbar surgery.

For wild natural medicine, unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials, which affects the efficacy and safety of clinical medication. DNA barcoding as an effective species identification tool is limited by its low sample throughput nature. In this study, combining DNA mini-barcode, DNA metabarcoding and species delimitation method, a novel biological sources consistency evaluation strategy was proposed, and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as "Guang Dilong" and 25 batches of proprietary Chinese medicines. Besides Amynthas aspergillum as the authentic source, 8 other Molecular Operational Taxonomic Units (MOTUs) were elucidated. Significantly, even the subgroups within A. aspergillum revealed here differ significantly on chemical compositions and biological activity. Fortunately, this biodiversity could be controlled when the collection was limited to designated areas, as proved by 2796 "decoction pieces" samples. This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control, and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.