Objective To evaluate the role of spinal Toll-like receptor 4 (TLR4) signaling pathway in the development of inflammatory pain in rats.Methods Thirty-six adult male Sprague-Dawley rats were divided into 3 groups (n=12 each) using a random number table:control group,inflammatory pain group and TLR4 signaling pathway inhibitor epigallocatechin gallate (EGCG) group (EGCG group).Inflammatory pain was induced by injecting 50 μl of complete Freund's adjuvant (CFA) into the ankle joint cavity of the left hindpaw of rats anesthetized with isoflurane.At 1-3 days after injection of CFA,EGCG 30 μg was intrathecally injected once a day in group EGCG.At 1,3 (30 min after intrathecal injection),5and 7 days after injection of CFA,the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured.The ipsilateral L4.5 segments of the spinal cord were removed at 3 days after CFA injcction for determination of TLR4 expression (by Western blot) and contents of tumor necrosis factor-alpha (TNF-oα),interleukin-6 (IL-6) and IL-1β in the spinal dorsal horn (by enzymelinked immunosorbent assay).Results Compared with control group,the MWT was significantly decreased and the TWL was shortened at each time point after injection of CFA,the expression of TLR4 in the spinal dorsal horn was up-regulated,and contents of TNF-α,IL-6 and IL-1β in the spinal dorsal horn were increased in inflammatory pain group (P＜ 0.05),and no significant change was found in the parameters mentioned above in EGCG group (P＞0.05).Compared with inflammatory pain group,the MWT was significantly increased and the TWL was prolonged at each time point after injection of CFA,the expression of TLR4 in the spinal dorsal horn was down-regulated,and contents of TNF-α,IL-6 and IL-1β in the spinal dorsal horn were decreased in EGCG group (P＜0.05).Conclusion Spinal TLR4 signaling pathway is involved in the development of inflammatory pain in rats.

How to effectively using the electronic health records (EHR) in contracted family doctors service was studied according to the new model of HER-based community medical service with community health service station as a center in combination with large hospitals,and suggestions were put forward for the solution of problems in contracted family doctor service,namely establishing and perfecting the community health service system,enlarging the scale of HER,strengthening the construction of general doctors team,and training the technically competent community health service staff.

Using the concept and method of evidence-based medicine,the paper systematically analyzes the functions and shortage of intelligent health Application (App) in disease diagnosis,treatment and self-management,and describes the functions of health and disease management App,so as to promote the development of App and enable App to better assist in health management.

AIM: To evaluate the influence of anesthesia and different means of postoperative pain control on the T-lymphocyte during the perioperative period in patients with rectal cancer.METHODS: 40 adult patients, aged 65 or older, of American Society of Anesthesiologists (ASA) class 2-3 were divided into two groups according to the type and means of postoperative pain managements. Group Ⅰ (n=20) received intravenous anesthesia and patient controlled analgesia(PCA), fentanyl (13 ?g/kg) for post pain; group Ⅱ (n=20) received intravenous anesthesia plus lumber epidural anesthesia and epidural PCA of morphine 5 mg plus ropivacaine 100 mg for post operative pain. Blood samples from internal jugular vein were obtained before surgery, at the completion of surgery and 24, 48, and 120 h post surgery for detecting CD3+, CD4+, CD4/CD8 counts of peripheral T-lymphocytes. In addition, blood cortisol level and pain intensity were assessed by visual analogue score (VAS)at each time point. RESULTS: Baseline(before anesthesia) values of CD3+,CD4+, CD4/CD8 in patients were messured and there was a significant decrease of all these values from completion of surgery to 48 h after surgery in both groups (P

Objective To evaluate the role of interleukin-12 (IL-12) in the spinal cord in the maintenance of arthritic pain (AP) in rats.Methods Adult male Sprague-Dawley rats, aged 6-8 weeks,weighing 200-300 g, were randomly divided into 4 groups using a random number table: control group (group C, n=6);AP group (n=9);phosphate buffer solution (PBS) group (n=6);IL-12 antibody group (n =6).AP was induced by injecting 50 μl of complete Freund' s adjuvant into the ankle joint cavity of the left hindpaw of rats anesthetized with isoflurane.Goat anti-rat IL-12 antibody 1.50 μg (20 μl) was intrathecally injected on 9 days after establishment of the model in group IL-12 antibody, while 0.01 mol/L PBS (20 μl) was administered in group PBS.The mechanical paw withdrawal threshold to yon Frey filament stimulation (MWT) was measured before establishment of the model (baseline) and on 9 and 10 days after establishment of the model.The rats were sacrificed after the last measurement of pain threshold, and the L4 6 segments of the spinal cord were obtained for detection of IL-12 expression in the spinal dorsal horn (by immunofluorescence) , and the co-expression of IL-12 and glial fibrillary acidic protein (an astrocyte marker) was examined simultaneously in group AP.Results Compared with group C, the MWT was significantly decreased, and the expression of IL-12 was up-regulated on 9 and 10 days after establishment of the model in AP, PBS and IL-12 antibody groups.Compared with group AP, the MWT was significantly increased at 10 days after establishment of the model, and the expression of IL-12 was down-regulated in group IL-12 antibody, and no significant change was found in the MWT at 10 days after establishment of the model and expression of IL-12 in group PBS.IL-12 was co-expressed with glial fibrillary acidic protein in group AP.Conclusion IL-12 in the spinal cord is involved in the maintenance of AP in rats.

Diabetes mellitus is a chronic disease, it is thus necessary for its patients to participate in its treatment and self-management. The emerging foreign and domestic methods for self-management of diabetes mellitus pa-tients were detected on different Websites and suggestions were proposed for its treatment and self-management in order to improve the quality of life in diabetes mellitus patients.

Objective:In this study,a series of experiments were conducted to research the mechanism of anticancer and preliminary molecular effects of PAMs on the HEPG-2 cancer cells.Methods:Morphological observation and MTT assay were used to explore the inhibition and killing effect of PAMs acting on HEPG-2.AO/EB staining and Annexin V-FITC/PI staining were employed to observe the apoptosis of HEPG-2 treated with PAMs.The expression level of Foxm1,bcl-2 and others genes in HEPG-2 cells were detected by using qRT-PCR and western blot.Wound healing and transwell experiments determined if PAMs can inhibit the migration of HEPG-2.Results:PAMs can inhibit and kill HEPG-2 cells in time and dose-dependent manners,and the cytotoxic effects were closely related to the cell apoptosis.The mRNA expression of foxm1,bcl-2 and surviving gene were remarkably decreased in HEPG-2 cells after the treatment of PAMs.PAMs decreased the FoXM1 protein expression in HEPG-2 cells,while up-regulating thep53 protein expression.,and it could also inhibit the migration of cancer cells.Conclusions:The possible molecular mechanism for the killing of HEPG-2 cancer cells by PAMs was proposed.By down-regulating the expression of foxm1 and up-regulating the expression of p53,the transcriptional expression of their downstream target genes survivin and bcl-2 was inhibited or reduced,hence enhancing the cancer cell apoptosis.This study provides an important foundation for the development of anti-cancer Chinese folk medicine based on PAMs.

AIM:To observe and analyze the effect of cardiopulmonary bypass ( CPB) for 30 min on surface ultra-structure and mechanical properties of the erythrocyte membrane by atomic force microscopy (AFM).METHODS:Ten cases of elective patients in cardiac surgery were selected in the study and divided into control ( CON) group and CPB group.The central venous blood (2 mL) before surgery and 30 min after CPB was collected with heparin anticoagulation . The non-circular red blood cells were counted under a stand fluorescence microscope .AFM was used to examine the ultra-structure of the membrane surface and measure the force curve of the erythrocytes .RESULTS:The percentage of non-cir-cular red blood cells in CPB group showed no statistically significant differences as compared with CON group .AFM images showed that the significant differences of membrane surface concave and convex , evenness , particle distribution , the sur-face average roughness (Ra), the surface root mean square roughness (Rq) and cell membrane adhesion between CPB group and CON group were observed .However, the membrane deformation resilience and curve slope had no significant difference between the 2 groups.CONCLUSION:Cardiopulmonary bypass for 30 min changes the morphology and ultra-structure of the erythrocyte membrane surface , and increases the adhesion between cells .

Objective To explore the application of endometriosis fertility index (EFI) in guidance after laparoscopic surgery of endometriosis patients combined with infertility and to explore methods to improve pregnancy rate in different EFI groups. Methods A prospective research was done in endometriosis patients combined with infertility in Beijing Obstetrics and Gynecology Hospital from January 2010 to June 2011, after laparoscopic surgery, these 146 patients were divided into 3 groups by EFI score. Using different pregnancy guidance, these patients had 5 years follow-up. Results (1) The 5 years overall pregnancy rate was 89.0%(130/146). The pregnancy rate was 95.7%(45/47) in EFI≥9 group, 92.8%(77/83) in EFI 5-8 group and 8/16 in EFI≤4 group, three groups were all reach satisfactory pregnancy rate;the rate of the first two groups had no statistically significance (P=0.498), but had significant difference with the last group (P<0.01). (2) In EFI≥5 patients, pregnancy rate was the highest in 6 months after operation;in EFI≥9 group, the pregnancy rate was 66.7%(30/45), and EFI 5-8 group was 50.6%(39/77). (3) EFI≥9 group had the highest natural pregnancy rate [83.6% (46/55)], natural pregnancy rate was significant statistical different in different EFI groups (P=0.001). Conclusions EFI score is a useful evaluation in predicting and guiding pregnancy in endometriosis patients combined with infertility after laparoscopic surgery. EFI score guidance, strict post-operation management and positive pregnancy scheme could significantly improve the pregnancy rate of endometriosis patients with infertility.

CRISPR/Cas9 technology originated from type II CRISPR/Cas system, which is widely found in bacteria and equips them with acquired immunity against viruses and plasmids. CRISPR-associated protein Cas9 is a RNA-guided endonuclease, which can efficiently introduce double-strand breaks at specific sites and activate homologous recombination and/or non-homologous end joining mechanism for the repair of impaired DNA. Features such as easy-to-use, cost-effectiveness, multiple targeting ability have made it the third-generation genomic engineering tool following ZFNs and TALENs. Here the history of discovery and molecular mechanism of the CRISPR/Cas9 technology are reviewed. The rapid advance in its various applications, especially for the treatment of human genetic disorders, as well as some concomitant problems are discussed.
Biomedical Research ; methods ; trends ; CRISPR-Cas Systems ; genetics ; Gene Editing ; methods ; Genome, Human ; genetics ; Humans ; Models, Genetic ; Reproducibility of Results