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 investigate the relationship between hypercoagulable state and the activation of the NF- kappa B pathway,inflammatory/suppression cytokines in patients with osteoarthritis. Methods:56 patients with OA were divided into two groups according to random number table:XFC group ( 28 cases ) and glucosamine ( GS ) group ( 28 cases ) . Two groups were treated for 3 months. Nine healthy people are healthy control group ( NC) . Determining the expression levels of the index of the NF-κB signaling pathway (p50,p65,TAK1,IκBα) and TNF-α,IL-1,IL-10,platelet activating factor(PAF) in serum by enzyme-linked immunosorbent assay. Detected the level of the indicators and laboratory indexes related with coagulation,observed the changes between the two group, used OA symptoms integral scale, LequesneMG, SF-36 and vas to assess efficacy;and made a correlation analysis. Results: After treatment,FIB,D-D, PAF, PLT, p50, p65, TAK1, IL-1, TNF-α, hs-CRP, ESR, IgG, LequesneMG, symptom integral meter, and VAS integral were significantly higher,APTT,PT,PAF-AH,IL-10 and each dimension integral of SF-36 significantly decreased in 2 groups (P<0. 05). XFC group was better than the GS group in reducing the level of PLT,FIB,TNF-α,p65,TAK1,hs-CRP,ESR,symptom integral meter,and VAS integral and increasing PT,each dimension integral of SF-36, etc (P<0. 05,P<0. 01). The results of Pearson correlation analysis show that PLT,FIB,D-D,PAF had positive correlation with p50,p65,TAK1,IL-1,TNF-α,hs-CRP,ESR,IgG,Le-quesneMG,symptom integral meter,and VAS integral,and negative correlation with IL-10 and each dimension integral of SF-36 ( P<0. 05,P<0. 01). PT had positive correlation with IL-10,GH and PF,and negative correlation with p50,p65,TAK1,IL-1,TNF-α,hs-CRP,ESR,IgG,symptom integral meter,and VAS integral (P<0. 05,P<0. 01). Conclusion: XFC could inhibit the NF-κB signaling pathway, raise the level of IL-10, reduce the expression of IL-1, TNF-α, P50, p65, TAK1 and so on, and reduce the abnormal inflammatory immune response. So as to achieve the purpose of delaying and inhibiting the production of hypercoagulable state,reduce joint disease,relieve the symptoms of joint pain and stiffness,eventually improve the patient’s quality of life.

Objectives:To understand the current situation of premenstrual syndrome (PMS) and type D personality in female college students,and to explore the relationship between them.Methods:Totally 856 female college students were assessed with the Premenstrual Syndrome Scale (PMSS,those with total score of 6 or higher as having PMS) and Type D Personality Scale 14 (DS-14,those negative emotion and social inhibition factor with total score of 10 or higher as having type D personality).Result:A total of 280 students were detected with PMS,and the rote was 32.7％.Totally 272 students were detected with the type D personality,and the rote was 31.8％.The total score of DS-14 and the score of 2 factors were higher in PMS group than in non-PMS group (Ps ＜0.001).The rate of PMS was lower in type D personality group than in non-type D personality group (11.76％ vs.25.34％,P ＜ 0.001).Conclusions:The occurrence of premenstrual syndrome may be related to the type D personality.The evaluation of personality characteristics should be taken into consideration when formulating the intervention program.

Objective:To observe the changes in coagulation parameters, peripheral blood cytokines, NF-kappa B signaling pathway protein, laboratory indexes in Sjogren′s syndrome ( SS ) patients, thus to explore the mechanism of hypercoagulable state. Methods:60 patients with SS and 20 healthy persons were randomly selected as the study group and the control group;Automatic coagulation analyzer was used to detect the value of coagulation parameters [ activated partial thromboplastin time ( APTT) ,prothrombin time ( PT) , plasma fibrinogen ( FIB ) , prothrombin time ( TT ) , D-dimer ( DD ) ]; ELISA method was performed to observe the expression of related cytokines(IL-1β,TNF-α,IL-10) and NF-κB signaling pathway proteins (p65,p50,IκBα); Westergren method was used to determine erythrocyte sedimentation rate ( ESR) ,and automatic biochemical analyzer to examine immune protein( IgG,IgA, IgM,GLO) and high sensitivity C-reactive protein (hs-CRP). Results: Blood coagulation parameters in 60 patients with SS were at least one abnormal for 46 cases,accounting for 76. 7% of the subjects. Among them,the abnormal rate of D-D was the highest,followed by FIB,APTT,PT,TT. Compared with the control group,D-D,FIB were significantly increased in SS patients,and TT,PT,APTT was not found obviously different. In addition,IL-1β,TNF-α,P50,P65,IκBα and inflammatory indexes like ESR,hs-CRP,IGg,GLO,ESSDAI, corneal staining score increased while the salivary flow rate,tear film break-up time and IL-10 decreased significantly( P<0. 05 or P<0. 01). Correlation analysis showed that the coagulation parameters FIB were positively correlated with the salivary flow rate,TNF-α, P50,P65,ESR,hs-CRP,while negatively correlated with IL-10,that TT was negatively correlated with TNF-α;that D-D was positively correlated with TNF-α, IL-1β, P65, ESR, hs-CRP, ESSDAI and corneal staining score, while negatively correlated with IL-10. Conclusion: SS patients generally have hypercoagulable state, and may be associated with the imbalance of cytokines, abnormal activation NF-kappa B signaling pathway,which mediates vascular endothelial cell damage,causing coagulation/fibrinolytic system dis-orders.

OBJECTIVE:To determine the content of phenolic acids from Anemone altica,and optimize its extraction technolo-gy. METHODS:HPLC was used to determine the contents of mono-ferulyl tartaric acid and ferulic acid from A. altica;using the total contents of 2 index components as index,volume fraction of extraction solvent,extraction solvent volume,extraction times and extraction time as factors,orthogonal test was used to optimize extraction technology,and verification test was conducted. RE-SULTS:The contents of mono-ferulyl tartaric acid and ferulic acid were 0.059%,0.0025%,respectively;the optimal extraction technology was as follow as 30% ethanol 600 mL added to 20 g medicinal material,extracted twice,90 min every time. In verifi-cation test,the average contents of 2 components in extract were 0.2971%(RSD=3.64%,n=3),0.0041%(RSD=5.11%,n=3). CONCLUSIONS:A method for contents determination of mono-ferulyl tartaric acid and ferulic acid from A. altica is estab-lished;optimized extraction technology is stable and feasible.

BACKGROUND:There is little evidence on the use of al ogeneic tendon graft in the reconstruction of ankle joint.
OBJECTIVE:To explore the clinical outcome of anatomical reconstruction of the lateral ligaments with cryopreserved al ogeneic tendon graft in patients with chronic ankle instability.
METHODS:Twenty-six patients with chronic lateral instability underwent anatomical reconstruction of the lateral ligaments with cryopreserved al ogeneic tendon. There were 18 cases of simultaneous injury or chalasia in calcaneofibular ligament and anterior talofibular ligament, and 8 cases of anterior talofibular ligament injury or chalasia. The ankle joint function was evaluated according to AOFAS scale and Good classification. The affect ankle and healthy ankle were compared in the extension, plantar flexion activity, and metaleg activity.
RESULTS AND CONCLUSION:Al the 26 patients were fol owed up for 9-24 months with a mean of 15 months. No cases appeared recurrent ankle lateral instability. The mean AOFAS score in the group of calcaneofibular ligament and anterior talofibular ligament was improved from (48.4±3.7) points preoperatively to (88.2±3.8) postoperatively, while that in the group of anterior talofibular ligament was improved from (50.0±6.4) points preoperatively to (89.5±3.4) points postoperatively. According to Good score, there were excellent in 19 feet, good in 6 feet, fair in 1 foot, with an excellent and good rate up to 96%. No serious complication was occurred in this group. Anatomical reconstruction of the lateral ligaments with cryopreserved al ogeneic tendon graft can increase the tendon-bone contact area, improve the rate of tendon healing, and enhance the stability of ankle joint in patients with chronic ankle instability. Further studies are needed to verify its long-term efficacy.

Objective:To explore the application of high-throughput sequencing (HTS) technology in pathogens detection for spinal infection.Methods:From January 2019 to May 2020, a total of 41 patients including 31 males and 10 females with an average age of 59.7±11.9 years (29-75 years) were suspected of spinal infections. There were 37 patients with local pain, 15 with fever (≥38 ℃) and 18 with neurological dysfunction. The infected sites were as follows, 4 cases of cervical spine, 8 cases of thoracic spine and 29 cases of lumbar spine. There were 36 patients met the surgical indications and underwent open debridement, bone grafting, fusion and internal fixation, while the other 5 patients underwent conservative treatment (three received drug therapy and two were transferred to the internal department for chemotherapy). Lesions obtained from open surgery patients were underwent pathology and HTS examination. In 5 cases with conservative treatment, two of them underwent CT guided percutaneous puncture for samples, while one case underwent ultrasound guided percutaneous puncture for pus, one case for venous blood, and one case received lumbar puncture for cerebrospinal fluid. The samples were sent for pathological and HTS examination, while liquid specimens were sent for bacterial culture and HTS. The sensitivity and specificity of HTS results were determined according to pathological examination which was regarded as the "gold standard". Based on HTS results combined with the clinical manifestations, imaging examination and pathological results of the patients, targeted antibiotics or anti-tuberculosis drugs were selected for postoperative drug therapy. Patients with bacterial infection received anti-infection treatment for 3 months after operation. For tuberculosis patients, "tetrad" (isoniazid+rifampicin+pyrazinamide+ethambutanol) anti-tuberculosis treatments were underwent for one year. Inflammation indicators from the blood samples were observed before and after treatment, including white blood cell count (WBC), C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). These indicators were used to monitor disease progression and the curative effects. All patients were followed up for at least 3 months after surgery.Results:A total of 41 patients with suspected spinal infection were included in this study. The HTS pathogen detection results were obtained within 48 h. For the initial 5 patients, first-generation sequencing verification was conducted with coincidence rate 100%. Further, no further verification was conducted in the rest patients. Among the 41 cases, a total of 26 cases had positive results with a positive rate of 63.4%(26/41). Among them, thirteen cases were with mycobacterium tuberculosis (31.7%) and 6 cases with staphylococcus (14.6%). Fungi and Brucellosis were diagnosed in 2 cases respectively, accounting for 4.9% respectively. The test were negative in 15 patients (36.6%), including 2 patients with tumor or tumor-like lesions (1 hematologic tumor and 1 eosinophilic granuloma). A total of 38 patients underwent pathological examination, which confirmed 7 cases of suppurative infection, 12 cases of tuberculosis, 2 cases of tumor or tumor-like lesions and the remaining 17 cases of inflammatory lesions. The sensitivity and specificity of HTS were 80%(16/20) and 55.6% (10/18) with positive predictive value (PPV) 66.7% (16/24) and negative predictive value (NPV) 71.4% (10/14). All patients were followed up for 3 months. The inflammation indicators of blood at 3 months were all lower than that at admission. WBC decreased from (7.50±3.26)×10 9/L at admission to (6.22±2.53)×10 9/L at 3 months after treatment without statistically significant difference ( t=1.082, P=0.290). The CRP decreased from (32.2±34.1) mg/L to (4.5±10.5) mg/L, and ESR from (44.2±26.5) mm/1 h to (18.6±12.1) mm/1 h with statistically significant difference ( t=8.963, P<0.001; t=5.421, P<0.001). Conclusion:High-throughput sequencing technology can be used in detection of spinal infection pathogens, due to its relatively high positive rate, satisfied sensitivity and good diagnostic value.

Objective To investigate the relationship between single nucleotide polymorphism in estrogen metabolizing genes CYP17、CYP19 and breast cancer susceptibility.Methods A case-control study was performed.PCR-base restriction fragment length polymorphism(PCR-RFLP)and short tandem repeat polymorphism(STRP)assay were used to detect the single nucleotide polymorphism of CYP17、CYP19 in 213 breast cancer cases and 430 matched controls.Resuits CYP17 A2/A2 genotype was found in 6.7%of breast cancer cases,which was significantly higher(P＜0.05)than that in controls(2.4%);the frequency of A2 allele of CYP17 was 16.2%in breast cancer cases,which was significantly higher(P＜0.05)than that in controls(10.6%).There Was alSO significant difference in the frequency of(TTTA)10allele of CYP19 which was 12.4%in breast cancer cases and 8.2%in controls(P=0.02).Conclusions The allele of CYP17 A2 and CYP19(TTTA)10 and CYP17 A2/A2 genotype were positively associated with the susceptibility of breast cancer.

Objective To investigate the expression of NMDAR1 in the hippocampus and cerebral cortex of old rats after 30-min -inhalation of 2% isoflurane, and to investigate the effects of isoflurane on the learning and memory functions of old rats and the underlying mechanism. Methods The healthy old male Sprague Dawley rats (n = 36) were randomly divided into the control group, the oxygen group, the 2-hour post-recovery group, the 1-day post-recovery group, the 3-day post-recovery group, and the 7-day post-recovery group. The morris water maze was used to detect the ethological effect of 30-min inhalation of isoflurane , and the immunohistochemistry assay was used to detect the expression of NMDAR1 in the hippocampus (CA1, CA3) and the cerebral cortex. Results The 30-min inhalation of 2% isoflurane inhibited the learning and memory abilities of old rats at 2 h post-recovery. On 1 d post-recovery, the inhibition of learning and memory began to reduce, then on 3 d and 7 d post-recovery, the learning and memory abilities continously recovered. The expression of NMDAR1 in the rat hippocampus and cerebral cortex decreased at 2 h post-recovery, and reversed on 1 d post-recovery and reached the normal level on 3 d and 7 d post-recovery. Conclusion 30-min inhalation of 2%isoflurane had an inhibitory effect on the learning and memory abilities of old rats, and the attenuation of NMDAR1 in the hippocampus and cerebral cortex may involve in this process.

Objective To investigate the risk factors and preventive strategies of patients with diffuse axonal injury(DAI) with deep veins thrombosis in lower limbs (LDVT).Methods One hundred and thirty cases of diffuse axonal injury patients with lower limb vascular were divided into LDVT group(22 cases) and non LDVT group(108 cases) based on ultrasound.The information including long-term bed,plasma fibrinogen level,varicose veins,hypertention,sex,age,smoking,alcohol drinking,diabetes,obesity,Glasgow Coma Scale (GCS) were collected.Results There were significant different between LDVT and non-LDVT group in terms of longterm bed time,hypertension,smoking,diabetes,high plasma fibrinogen,age,low GCS score correlated with LDVT (x2 =7.08,5.99,5.17,4.70,3.55,12.72,t =27.80,P ＜ 0.05).Gender,drinking,obesity,varicose vein factors had no correlation with LDVT(P ＞ 0.05).Conclusion Diffuse axonal injury in patients with LDVT is more common in patients with older age,hypertension,low GCS score,the higher the plasma fibrinogen.