Objective To explore the effect of fibroblast growth factor receptors 1-dominant negative strategy (FGFR1-DN) on alkaline phosphatase (ALP) activity of bone marrow stromal stem cells (BMSCs) after osteogenic induction.Methods BMSCs were transfected with eukaryotic expression plasmid pcDNA 3.1 (+)-DN FGFR1 and pcDNA3.1 (+)-FGFR1.The experiment was conducted in 4 groups:FGFR1-DN transfection group,FGFR1 transfection group,pcDNA3.1(+) empty vector transfection group and non-transfection group.The ALP activity of BMSCs was detected in logarithmic growth phase after osteogenic culture.The qualitative detection of ALP activity was carried out immunohistochemically while the quantitative detection by cALP kit.The ALP activity was compared between the 4 groups at 7 and 14 days after osteogenic induction.Results Compared with 7 days,the ALP activity at 14 days was significantly increased in the 4 groups,and the increase in FGFR1-DN transfection group was significantly higher than in the other 3 groups (P ＜ 0.05).At both 7 and 14 days,the ALP activity in FGFR1-DN transfection group was the highest while that in FGFR1 transfection group was the lowest (P ＜ 0.05).Conclusions FGFR1-DN can promote the ALP activity of BMSCs during osteogenesis.This may provide an experimental basis for the joint application of local gene therapy and tissue engineering and for construction of tissue engineered bone with better biocompatibility.

OBJECTIVE:To provide reference for further improving negotiation mechanism of drug medical insurance in Chi-na. METHODS:Questionnaire investigation was conducted among the medical insurance agency and pharmaceutical company staff who participated in drug medical insurance negotiation in Jiangsu province,Jiangxi province,Qingdao,Chengdu. Their business fa-miliarity and recognition of the negotiating mechanism were also investigated. The descriptive analysis,independent sample t test and other methods were adopted to analyze the data statistically. RESULTS:A total of 70 questionnaires were distributed and 68 val-id questionnaires were collected with effective recovery rate of 97.14%. Respondents included 21 staff members of medical insur-ance agencies and 47 of pharmaceutical companies. In terms of business familiarity,the average score of respondents in all aspects was greater than 3(4 points system),the lowest score wasmanagement of the implementation of drug negotiation projects. There were statistical significances in scores of 3 items between 2 parties in the negotiation(P<0.05):basic principles of the drug negotiation process should be followednegotiation objectives in the substantive negotiation process total score of busi-ness familiarity. The scores of respondents in pharmaceutical companies were higher than those of health insurance agencies. In the recognition of the negotiating mechanism,average score of each survey item was ≥4(5 points system)in the recognition of the overall effect of the negotiation mechanism except forwhich is conducive to maintaining the price system of pharmaceutical com-panies. The average score of all aspects of preparation of the negotiations,substantive negotiation,implementation of the negotia-tion agreement and negotiation project's supervision and evaluation was >4 except forpatients taking drugs is quick and easy. There was no statistical significance between 2 types of respondents in all aspects of overall effect of the negotiation mechanism, preparation of the negotiations and negotiation project's supervision and evaluation(P>0.05). But in the recognition of the substan-tive negotiation,there was statistically significance between 2 types of respondents innegotiations between the two sides in an equal,cooperative,harmonious atmosphere(P<0.05). Healthcare agencies'respondents had higher scores than pharmaceutical companies'ones. And in the recognition of the implementation of the negotiation agreement,there was statistical significance be-tween 2 types of respondents in theafter the agreement entered into force,the medical insurance agency will be strictly in accor-dance with the agreementandfund pay to pharmaceutical companies timely and reasonable(P≤0.05). Healthcare agencies're-spondents have higher scores for than pharmaceutical companies'ones. CONCLUSIONS:The current drug medical insurance nego-tiation mechanism shows good overall effect and runs smoothly. But there are still some shortages,such as collected information the preparation stage is not sufficient;the procedures of patients taking the drug is complicated;negotiating parties do not have equal status;the fund is not paid to the pharmaceutical enterprise timely;evaluation is not perfect.

Objective To analyze the clinical characteristics and prognostic factors in patients with primary gastrointestinal diffuse large B-cell lymphoma (PGI-DLBCL),and to improve the diagnosis and treatment of PGI-DLBCL.Methods Retrospective analysis was conducted in 51 cases of PGI-DLBCL between January 2009 and August 2013.The data included clinical manifestations,pathological features,treatment regimens and prognosis.Results 51 patients included 31 males and 20 females,the range of ages was from 16 to 80 years old,median age was 48 years old.The major clinical presentation were abdominal pain,abdominal distension,abdominal mass,nausea and vomiting,abdominal mass.The occurrences in stomach,small intestine,colon,rectum and multiple involvement were 56.86 ％,29.41％,7.84 ％,1.90 ％ and 3.92 ％ respectively.The mass bigger than 10 cm was found in 13 cases (25.49 ％).47.06 ％ (24/51) of the cases belonged to the GCB subtype and 52.94 ％ (27/51) belonged to the non-GCB subtype.There was no significant impact of lymphoma cell origin,disease distribution (stomach or intestinal) and mass on prognosis of lymphoma treatment.The univariate analysis revealed that the patients with Lugano stage Ⅳ,increased level of serum lactate dehydrogenase (LDH),modified-international prognosis index (modified IPI) 3-5 and increased level of CA125 had poor prognosis (all P ＜ 0.05).There was no difference of survival rate between patients treated with rituximab plus chemotherapy and single CHOP like therapy.Surgery plus postoperative chemotherapy significantly improved survival of patients treated with simple chemotherapy (P ＞ 0.05).Conclusion The clinical Lugano stage,IPI score,increased LDH and CA125 are important prognostic factors of PGI-DLBCL.

Objective To investigate the effect of nucleocapsid (N) protein of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) on the expression of cyclooxygenase-2 (COX-2). Methods 293T cells were co-transfected with reporter plasmid pCOX-2-Luc containing the luciferase gene under the control of COX-2 promoter and plasmids carrying individual genes of SARS-CoV, and luciferase activity was measured. Expression of COX-2 mRNA and protein was measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Results N protein of SARS-CoV enhanced COX-2 gene promoter activity, and upregulated COX-2 mRNA expression.COX-2 protein production in 293T cells was N protein concentration-dependent. Conclusion N protein of SARS-CoV could specifically activate COX-2 expression.

Objective To investigate the effects of the point-injection combined with neuromuscular facilitation rehabilitation techniques on post-stroke shoulder-hand syndrome. Methods A treatment group, 36 cases, was treated with the point-injection combined with neural facilitation of rehabilitation techniques. And a control group, 30 cases, was treated with massage therapy. Observed the clinical manifestations and used Fugl-Meyer to assess the joint activities, pain degree and motion function of upper limbs before and after therapy. Results Compared with the control group, the treatment group showed better improvement of joint activity scope and degree, and alleviation of pain (P<0.05). Conclusion Point injection and neuromuscular rehabilitation treatment has a better effect ain treating sequelare of brain stroke and it is worth applying.

BACKGROUND:Interlocking intramedullary nailing is a main method for bone fractures,but traditional static intramedullary nails usually lead to nail breakage and loosening.Thereafter,we design a novel high-thread wooden club-shaped screw (HTWCSS) and explore its mechanical properties.OBJECTIVE:To measure the bend strengths of HTWCSS,analyze its stress distribution,and to evaluate its biomechanical properties,thereby providing theoretical basis for its clinical application.METHODS:The bend strength of HTWCSS and traditional nails were measured via three-point bending experiments.Transverse fractures of the middle tibia were simulated using finite element method,and then the force and stress distribution of the two different nails were analyzed.RESULTS AND CONCLUSION:(1) The average maximum load of HTWCSS was 3.52 kN and 1.81 kN at span of 20 mm and 30 mm,respectively,which were larger than those of the traditional nails.(2) The average maximum displacement and the stress of HTWCSS were smaller than those of the traditional screws in finite element analysis,and the stress distribution was relatively dispersed.(3) The average maximum axial displacement and stress HTWCSS,traditional interlocking screws,wooden club-shaped screws and traditional screws were 131 MPa and 3.27 mm,162 MPa and 4.07 mm,26.5 MPa and 0.323 mm,and 34.3 MPa and 0.407 mm,respectively.(4) These results suggest that HTWCSS has relative high bend strength,and it is able to disperse stress and improve fatigue strength bend strength,further reduce screw broken.

Objective To study the protective effect of hyperin( Hyp) on the acute liver injury in rats induced by CCl4 . Methods The acute liver injury model was induced by CCl4 . The effect of Hyp on acute liver injury of rat liver histopathology was observed;and the impact of changes of homogenates total superoxide dismutase ( T-SOD) ,glutathione ( GSH) activity and malondialdehyde ( MDA) in liver were de-tected. Results The acute liver injury of tissue induced by CCl4,apparent pathological inflammation and fibrous tissue degeneration and necrosis were observed by HE staining;At the high doses of 60 mg/kg and medium doses of 30 mg/kg treated by Hyp,liver pathology changes was significantly obvious. The T-SOD,GSH activity of liver tissue was significantly increased in the groups of Hyp treatment,MDA content was significantly decreased,which related to the dosages. Conclusion In the rat model of acute liver injury induced by CCl4 . A better therapeu-tic effect of Hyp was shown,the mechanism may be related to its antioxidant activity.

Objective To investigate the mineralization impact of fibroblast growth factor receptors 1 dominant negative (FGFR1DN) on osteogenic induction culture of bone marrow stromal cells (BMSCs).Methods The 3rd generation BMSCs were divided into 4 equal groups (n =24).FGFR1-DN group was transfected by pcDNA3.1 (+)-FGFR1DN,FGFRI group by pcDNA3.1 (+)-FRFR1,blank load group by pcDNA3.1 (+)-blank vehicle and non-transtection group by nothing.After successful transfection was confirmed when the cells were in the logarithmic phase,osteogenic induction culture was conducted continuously for 21 days.Mineralized nodule formation was observed by alizarin red staining.The amount of mineralized material was calculated according to the standard curve of alizarin red concentration.Results Continuous osteogenic induction for 21 days showed on the bottom of the hole visible round opaque calcified nodules after alizarin red staining.The BMSCs in the FGFR1-DN group induced in the logarithmic growth phase by osteoblasts exhibited significantly increased osteogenic capacity while those in the FGFR1 group displayed diminished osteogenic capacity.The concentration of alizarin red was the highest in the FGFRI-DN group (1.33 ±0.19),the lowest in the FGFRI group (1.00 ± 1.17),and moderate in the blank load group (1.20 ± 0.16) and non-transfection group (1.17 ± 0.17),showing significant between-group differences (P ＜ O.05).Conclusions FGFR1-DN can promote cell proliferation in the early differentiation of BMSCs and also mineralization of osteoblasts in bone induction culture after the logarithmic growth phase.This may provide a hint for local gene treatment of bone defects.

Objective To analyze the urine trace albumin(mALb)/creatinine(Cr) ratio and blood uric acid(UA),and other various metabolic index level in patients with diabetic nephropathy(DN),combined with clinical data such as patients' age,body mass index(BMI),course of diseases,to explore the related mechanism of occurrence and development of DN.Methods 76 DN patients were selected.The microalbuminuria group(urinary mALb/Cr<300μg/mg) had 46 cases,the clinical albuminuria group(urinary mALb/Cr≥300μg/mg) included 30 cases,another 49 diabetic patients without kidney damage were seleted as control group.The urinary mALb/Cr,blood UA,fasting blood glucose(FBG),triacylglycerol(TG),total cholesterol(TC),high-density lipoprotein(HDL),low density lipoprotein(LDL),glycosylated hemoglobin(HbA1c) levels were determined.The BMI and the length of the course of the disease calculate.Results The patients' age,course of the disease,urinary mALb/Cr,blood UA,FBG,TC,TG,LDL,HbA1c and BMI level in the clinical albuminuria group and microalbuminuria group were significantly higher than those in the control group,the differences were statistically significant (F=6.18,12.48,141.43,12.48,8.49,4.98,6.18,3.89,3.17,3.89,all P<0.05).The high uric acid hematic disease rates of the clinical albuminuria group and microalbuminuria group were 26.09% and 26.09%,which were significantly higher than 10.20% of the control group,the differences were statistically significant(x2=4.074,24.833,all P<0.05).Urinary mALb/Cr was positively correlated with age,duration,BMI,UA,TG,TC,LDL,FBG,HbA1c(r=0.120,0.299,0.148,0.340,0.157,0.149,0.103,0.487,0.103).Multiple linear stepwise regression analysis suggested that duration,blood UA,FBG were independent risk factors of urinary mALb/Cr;TG,BMI,urinary mALb/Cr were independent risk factors for blood UA.Conclusion Urinary mALb/Cr and blood UA are the independent risk factors,high uric acid hematic disease may participate in the development process of DN,and diabetes duration,UA,BMI,TG,TC,LDL,FBG,HbA1C associated with increased urinary mALb/Cr excretory DN patients,the effective monitoring can improve the symptoms of DN and quality of life.

Objective:To investigate the short-term efficacy of anatomical calcaneal external fixator in the treatment of calcaneal fractures.Methods:A retrospective study was conducted of the 9 patients with calcaneal fracture who had been admitted to Department of Orthopedics, 920 Hospital of the PLA Joint Logistics Support Force between September 2019 and March 2020. There were 7 males and 2 females, aged from 23 to 66 years (mean, 41.3 years). The injury was caused by a fall from a height in 6 cases and a traffic accident in 3 cases. By the Sanders classification, there were 2 cases of type ⅡB, 2 cases of type ⅢAC, 3 type Ⅲ AB, one of type Ⅲ BC and one type Ⅳ. By the AO grading for closed soft tissue injury in the 8 closed fractures, there were 5 cases of grade Ⅰ and 3 cases of grade Ⅱ. The one open fracture was type ⅢB by the Gustilo-Anderson classification and treated with skin flap repair after debridement. All the 9 patients were treated with an anatomic calcaneal external fixator during stage-one or stage-two surgery (8 patients in stage-one and one in stage-two). The fracture union time for the patients was recorded; the ankle and hindfoot score of American Orthopedic Foot and Ankle Society (AOFAS) and the protocol of Association for the Study and Application of the Method of Ilizarov (ASAMI) were used at the last follow-up to assess the short-term efficacy.Results:Postoperative follow-up for the 9 patients ranged from 6 to 18 months (mean, 9.8 months). The duration of fracture union in the 9 patients ranged from 2 to 4 months (mean 2.8 months). The AOFAS ankle and hindfoot score at last follow-up ranged from 79 to 93 points (mean, 84.7 points) for the male patients (6 excellent and one good), and from 82 to 91 points (mean, 86.5 points) for the female patients (one excellent and one good). According to the ASAMI protocol at last follow-up, the short-term efficacy was rated as excellent in 6 cases, as good in 2 cases and as fair in one. The patients with closed fracture could get out of bed for rehabilitation on day 3 after surgery. None of the patients experienced such complications as pin tract infection and osteomyelitis during follow-up.Conclusion:The anatomic external calcaneal fixator demonstrates fine short-term efficacy in the treatment of calcaneal fractures, due to its advantages like anatomical configuration, a limited number of implants, and a possibility of partial weight-bearing for exercises on the third postoperative day.