Objective Human immunoglobulin combinatorial library was generated by using phage surface display expression system, and phage antibodies (Fabs) to platelet were screened.Methods:PBMC were separated from a patient who Were transfusion refractory to platelet. mRNA was isolated and cDNA was synthesized by reverse transcriptase.The immunoglobulin heavy chain Fd and the light chain ? genes were amplified by half nested PCR and the PCR products were cloned into the expression vector pCome3 respectively.The immunoglobulin combinatorial library was constructed and screened by 3 rounds of affinity selection.Results Human Immunoglobulin Combinatorial Library was successfully constructed.The specific phage antibodies were highly enriched after 3 rounds of biopanning selection against platelet membrane proteins.Conclusion The antibody library and human monoclonal antibodies to platelet may be useful as molecular tools to study the anti platelet drugs, platelet related diseases, epitope of human platelet antigens.

Objective To analyze the status and the distribution of causes of injuries as well as disease burden in children under 5 years old in Sichuan Province.Methods The death information cards of children who died because of injuries in Sichuan Province in 2012 were analyzed,and the years of potential life lost and working years of potential life lost were calculated to evaluate the disease burden.Results 22.87％ (1 502/6 567) of children lost their lives because of injuries,the mortality of injury was 4.13‰,63.45％ of children died at the age between l and 4,and most of the children died at home or on the road; The top 3 causes of children death of injuries below 5 years old were drowning,accidental suffocation and traffic accidents.The years of potential life lost and working years of potential life lost were 73.05 years and 58.30 years.Conclusion Injury is one of the most mainly risk factor of health and life of children,the government should further prevent and control children injuries.

Objective:To have an experimental investigation on teaching reform of Orthopedics of TCM based on network resources.Method:Two comparison groups were set and observed when we were teaching the orthopedics students of 2003 grade.One was taught by traditional problem-based learning(PBL) method,the other was taught by PBL method based on network resources.Results:The creative thought and research ability of the students who had been taught by PBL method based on network resources were greatly improved.Conclusion:The teaching method based on network resources has so many advantages such as students being able to self-learn and research without time and space limit that it can be used more in medical students teaching.

AIM:To investigate the activation of nuclear factor ?B(NF-?B) induced by lipopolysaccharides(LPS) in rat alveolar macrophages(AMs) and its regulatory role in tumor necrosis factor(TNF-?) secretion.METHODS:The dynamic activity changes of NF-?B induced by LPS were determined with electrophoretic mobility shift assay(EMSA).Antisense oligonucleotides of NF-?B subunit(p65) were transfected into AMs prior to LPS stimulation.The effect of antisense oligonucleotide transfection on expressions of p65 and TNF-? in supernatant were measured with Western blotting and enzyme linked immunosorbent assay(ELISA),respectively.RESULTS:NF-?B activity increased markedly and reached its peak level at 4 h after LPS stimulation.After transfected with antisense oligonucleotides of NF-?B subunit(p65),expression of p65 and TNF-? in supernatant decreased markedly.CONCLUSION:NF-?B activity has a positive effect on regulating secretion of TNF-? in AMs induced by LPS.

Objective To analyze the maternal mortality in Sichuan province from 2015 to 2016 for the purpose of providing reference for the department of health and family planning,medical and health care institutions to take measures to reduce maternal mortality.Methods The death assessment panel reviewed 163 cases of maternal mortality data in Sichuan province from 2015 to 2016 in accordance with "twelve grid method" of the World Health Organization(WHO).The findings were descriptively analyzed.Results Provincial level had completed 163 cases of maternal death review,and it suggested that obstetric hemorrhage,pregnancy induced hypertension,venous thrombosis and pulmonary embolism,heart disease,amniotic fluid embolism were the five leading causes of maternal mortality in Sichuan province.Among the 163 evaluated death cases,death could be avoided in 124 cases(76.07％) and death was unavoidable in 39 cases(23.93 ％).According to the first influencing factors,among 124 cases of avoidable deaths in medical records,the death of 67 cases(54.03％) was caused by medical staff knowledge and skills,ranking first,including the county(37.90％) and township(5.65％);the death of 57 cases was caused by the personal family,accounting for 45.97％.Conclusion It's important for the reduction of maternal mortality in Sichuan province to strengthen standardized management of high-risk pregnant women,ensure the timely supply of blood resources in remote areas,and enhance the training of emergency medical knowledge for obstetric medical personnel.

Objective:To analyze the current situation and requirements of education for health technicians in maternal and child health care institutions, and put forward feasible strategies and measures to improve the comprehensive quality and professional level of the talent team of maternal and child health care institutions.Methods:Questionnaire survey was carried out on education needs of health technical staff of 11 maternal and child health care hospitals in 4 provinces (regions), and provincial, municipal and district-level medical institutions. The survey results were recorded by Epidata 3.1. SPSS 22.0 software was used for statistical analysis.Results:A total of 1 678 questionnaires were included in the analysis. A total of 1 313 people received training, accounting for 78.2%. The main reason for not receiving training was that the unit didn't arrange (180 people), accounting for 49.3%(180/365). There were 779 people who had more than 3 days of training, accounting for 59.3%. There were 384 people who were trained in superior general hospitals, accounting for 29.2%, and 268 people were trained in superior maternal and child health institutions, accounting for 20.4%. There were 837 people who learned the content of new professional progress, accounting for 50.8%(837/1 648). According to the interview, there were still some requirements for thematic training, further education, online learning, continuing education and standardized training.Conclusion:Maternal and child health care institutions have accelerated the construction of professional personnel, intensified training, and thoroughly implemented health personnel training programs, established a long-term mechanism, increased funding, improved training content, ensured the quality of training, and made a good job in hierarchical training to meet the learning needs of personnel at all levels. This is of great significance for strengthening the technical personnel of maternal and child health care institutions and improving their service capacity.

Objective:To reduce the immunogenicity of vaccinia virus vector by replacing the D8L region, which is a neutralizing antibody epitope in vaccinia virus, with an exogenous gene.Methods:A gene fragment encoding influenza virus hemagglutinin (HA) was inserted into the D8L region to replace it using homologous recombination technique. Then, a recombinant vaccinia virus influenza vaccine was constricted. A recombinant vaccinia virus vaccine with the TK region expressing HA was used as a control. The expression of HA was validated by Western blot. BALB/c mice were immunized with the vaccines and the serum antibody titers two weeks after each immunization were evaluated by ELISA and hemagglutination inhibition assay. The protective efficacy of the recombinant vaccinia virus was assessed through a challenge experiment.Results:Western blot confirmed the successful expression of HAD8L protein in the constructed recombinant vaccines. ELISA and hemagglutination inhibition assay showed that after the primary immunization, the anti-HA antibody titer induced by the recombinant vaccinia virus with D8L region mutation was slightly higher than that induced by the vaccine with TK region mutation, and the difference was statistically significant with the increase of immunization times ( P<0.05). The recombinant vaccinia virus with D8L region mutation showed significantly lower immunogenicity than the recombinant virus with TK region mutation after the primary immunization, but there was no significant difference between them with the increase of immunization times ( P>0.05). After H1N1pdm challenge, no virus was detected in the mice immunized with the recombinant vaccinia virus with D8L region mutation and the mice showed mild lung inflammation and less tissue damage. Conclusions:This study indicated that inserting exogenous genes into the D8L region of the neutralizing antibody epitope in the vaccinia virus vector could help to reduce the immunogenicity of the vector itself and enhance the immunogenicity of the exogenous genes. This provided a reference for the use of the vaccinia virus vector as a delivery tool in the field of vaccines or gene therapy.

ObjectiveTo observe the effect of bee acupuncture therapy on clinical symptoms and signs， as well as the level of inflammatory cytokine interleukin-6 （IL-6） in synovial fluid of patients with knee osteoarthritis. MethodsThe 94 patients with knee osteoarthritis were divided into the control group and the trial group by the random number table method， with 47 cases in each group. Both groups were given one tablet （60 mg） of etoricoxib orally every morning for 2 weeks. The control group also received microneedle shallow acupuncture therapy， once a day for 5 consecutive times followed 2-day pause， and continued 5 consecutive times， as a course of treatment； the trial group was treated with bee acupuncture therapy once every 2 days， 2 times a week， and 4 times as a course of treatment. Both groups have a course of treatment for 2 weeks. The changes in clinical symptoms and signs of patients in the two groups were observed and evaluated before treatment， after 1- and 2-week treatment， and 12-week follow-up and the differences in Lequesne index scores， HSS scores， Visual Analogue Scale （VAS） scores and IL-6 levels in knee synovial fluid between the two groups of patients were also compared. ResultsNo patients lost to follow up in either group. The Lequesne index scores and VAS scores were lower， and the HSS scores were higher in both groups at all time points after treatment compared with those before treatment （P＜0.05）. Compared at the same time after treatment， the Lequesne index scores and VAS scores of the trial group were lower than those of the control group， and the HSS scores were higher than those of the control group （P＜0.05）. IL-6 in synovial fluid was lower in the trial group at the 12-week follow-up than before treatment （P＜0.05）， but there was no significant difference between two groups at each time point（P＞0.05）. ConclusionBee acupuncture therapy for knee osteoarthritis can significantly improve clinical signs and symptoms， but has no significant effect on the level of IL-6 in knee synovial fluid.

The protein kinase B(Akt)pathway can regulate the growth,proliferation,and metabolism of tumor cells and stem cells through the activation of multiple downstream target genes,thus affecting the development and treatment of a range of diseases.Thioesterase superfamily member 4(THEM4),a member of the thioesterase superfamily,is one of the Akt kinase-binding proteins.Some studies on the mechanism of cancers and other diseases have shown that THEM4 binds to Akt to regulate its phosphorylation.Initially,THEM4 was considered an endogenous inhibitor of Akt,which can inhibit the phosphorylation of Akt in diseases such as lung cancer,pancreatic cancer,and liver cancer,but subsequently,THEM4 was shown to promote the proliferation of tumor cells by positively regulating Akt activity in breast cancer and nasopharyngeal carcinoma,which contradicts previous findings.Considering these two distinct views,this review summarizes the important roles of THEM4 in the Akt pathway,focusing on THEM4 as an Akt-binding protein and its regulatory relationship with Akt phosphorylation in various diseases,especially cancer.This work provides a better understanding of the roles of THEM4 combined with Akt in the treatment of diseases.

Da Qinjiaotang is a common classical prescription for the treatment of stroke. It originates from Collection of Writings on the Mechanism of Disease， Suitability of Qi， and the Safeguarding of Life as Discussed in the Basic Questions （《素问病机气宜保命集》） by physician LIU Wansu， and is composed of Gentianae Macrophyllae Radix， Glycyrrhizae Radix et Rhizoma， Chuanxiong Rhizoma， Angelicae Sinensis Radix， Paeoniae Radix Alba， Asari Radix et Rhizoma， Notopterygii Rhizoma et Radix， Saposhnikoviae Radix， Scutellariae Radix， Gypsum Fibrosum， Angelicae Dahuricae Radix， Atractylodis Macrocephalae Rhizoma， Rehmanniae Radix， Rehmanniae Radix Praeparata， Poria， and Angelicae Pubescentis Radix. Doctors of all dynasties have disputed the composition principle of the prescription and argued whether its treatment of stroke belongs to the theory of "internal wind" or "external wind". Through collating and analyzing ancient and modern literature related to the indications of Da Qinjiaotang， this paper was dedicated to the origin of syndrome differentiation and treatment of Da Qinjiaotang. According to LIU Wansu's original works， Da Qinjiaotang is a prescription for the treatment of "internal wind"， and in the prescription， wind medicinal herbs such as Gentianae Macrophyllae Radix， Notopterygii Rhizoma et Radix and Angelicae Pubescentis Radix removes stagnation， clears sweat pore， and makes qi and blood channels flow smoothly. However， later generations， affected by the idea of "external wind"， believe that this prescription is used for the treatment of "external wind". Ancient physicians gradually supplemented the symptoms of stroke， such as wry eye and mouth， hemibody pain and limb numbness， which were treated by Da Qinjiaotang， and Da Qinjiaotang was also applied to the treatment of other diseases， such as tendon dryness， convulsion and arthralgia. Modern doctors still explain the disease pathogenesis from the theory of "external wind" as deficiency in channels and collaterals and the entry of pathogenic wind， and the prescription has the effect of dispersing wind， clearing heat and nourishing and activating blood. In clinical practice， Da Qinjiaotang is mainly used to treat cerebrovascular diseases and peripheral facial paralysis in nervous system diseases， gouty arthritis and rheumatic arthritis in the rheumatic immune system and skin diseases. The above findings facilitate the research and development of Da Qinjiaotang.