Objective To study the influence of prothrombin complex concentrates (PCC) containing different levels of antithrombin (AT) and heparin on its procoagulation and anticoagulation activities.Methods PCC containing traces of heparin and AT were prepared.The design of orthogonal experiment: set AT and heparin two variable factors, each factor respectively from three levels: 0, 0.4, 0.8 IU/mL; 0, 2.5, 5.0 IU/mL.Their influence on the procoagulation and anticoagulation activities of PCC were analysed.Results The R value of the influence of AT on thrombin inhibitory capacities was higher than that of heparin.With the increase of AT level, time to peak gradually extended.Lag time of 0.4 IU/mL and 0.8 IU/mL AT was the same.The peak height of 0.4 IU/mL AT was highest.The higher the level of AT, the greater the impact on thrombin inhibitory capacities.0.4 IU/mL and 0.8 IU/mL AT had similar impact on thrombin inhibitory capacities.The R values of the influence of heparin on thrombin generation were higher than that of AT.With the increase of heparin level, lag time and time to peak were longer while the peak height declined sharply.0 IU/mL and 2.5 IU/mL heparin had similar influence on thrombin generation.The higher the level of heparin, the greater influence on thrombin inhibitory capacities.2.5 IU/mL and 5.0 IU/mL heparin had similar influence on thrombin inhibitory capacities.Conclusion AT has greater influence on the anticoagulation activities of PCC than that of heparin, while heparin has greater influence on the procoagulation activities of PCC than that of AT.PCC containing medium levels of AT and heparin is relatively balanced between procoagulation and anticoagulation.

OBJECTIVE: To explore the clinical effects of using custom-made tumor prosthesis replacement to treat malignant tumor located in the distal femur or proximal tibia. METHODS: A retrospective study of 29 cases malignant tumors around knee treated surgically from June 2001 to October 2008. In the study there were 19 males and 10 females, aging from 17 to 65 years, with an average of 38.5 years. The location of tumors was at distal femur in 22 cases and 7 cases at proximal tibia. After the tumor was extensively resected or radically resected, custom-made tumor prosthesis replacement was performed for reconstruction, simultaneously, neoadjuvant chemotherapy and radioactive-therapy were used in treating the tumor according the pathology. RESULTS: All patients were followed-up for 18-84 months, with average follow-up period of 54.2 months. The 3 year survival rate was 79.8%, and the 5 year survival rate was 63%. As for the complications, prosthesis breakage occurred in 2 patients, aseptic loosening in 1 patients, 2 had a recurrence of the soft tissue tumor for which the diseased limb amputation was performed. The mean MSTS score showed excellent limb function in 15 patients, good in 8 patients, fair in 5 patients and poor in 1 patient. The overall excellent and good function was obtained in 79% of the patients. No superficial or deep infection occurred. Two patients suffered common peroneal nerve injury, but were cured by symptomatic treatments. CONCLUSION: Application of tumor prosthesis can give a satisfactory functional outcome, which is an effect method in limb salvage treatment of malignant tumor around the knee.

Objective To explore the role of p16 gene methylation in fibroblasts in the occurrence and development of keloid.Methods Skin tissue specimens were resected from the lesions of patients with keloid and normal skin of healthy human controls.Fibroblasts were isolated from these tissue specimens and subjected a primary culture.An immunohistochemical analysis was performed to measure the expression of p16 protein in tissue specimens,real-time fluorescence-based quantitative PCR to determine the mRNA expression level (expressed as 2-△△Ct) of p 16 and DNA methyltransferases (DNMTs) in fibmblasts,and bisulfite sequencing PCR (BSP) to estimate the methylation status of p16 gene in the tissue specimens and primary fibroblasts.Results The keloid fibroblasts (KFbs) showed significandy lower mRNA expression of p16 gene (0.64 ± 0.18 vs.1.92 ± 0.23,t =10.54,P＜ 0.05),but significantly higher mRNA expressions of 3 DNMTs (DNMT1:2.58 ± 0.23 vs.1.13 ± 0.21,t =11.22,P ＜ 0.05; DNMT3A:4.87 ± 0.46 vs.2.38 ± 0.32,t =10.81,P＜ 0.05; DNMT3B:1.57 ± 0.12 vs.0.57 ± 0.16,t =12.45,P＜ 0.05) compared with the normal fibmblasts (NFbs).The DNA methylation rate in the p16 gene promoter region was significantly increased in keloid tissue (1.81％ ± 0.46％) and KFbs (3.15％ ± 0.94％) compared with normal skin tissue (0.90％ ± 0.35％,F =14.23,P＜ 0.01) and NFbs (0.17％ ± 0.29％,F=37.62,P＜ 0.01).Conclusions The methylation and low expression of p16 gene in KFbs may be associated with the uncontrolled growth of keloid,and DNMTs may play a role in the pathogenesis of keloid.

Objective To observe the effect of preoperative oral administration of carbohydrate on blood glucose,insulin resistance(IR) and inflammatory reaction after gastrointestinal operation.Methods 48 patients receiving gastrointestinal operation were randomly divided into the study group(n =23)and the control group(n =25).Patients in the study group were orally given 25％ glucose solution 300 ml 3 hours before operation.Before anesthesia induction,gastric contents were aspirated through nasogastric tube to examine its volume and pH.Serum high sensitivity C-reactive protein(hsCRP),fasting blood glucose,insulin level and homeostasis model assessment-insulin resistance(HOMA-IR) were detected before operation and on the first morning after operation between the two groups.Results No anesthesia or operation related complications occurred in either groups.Patients had similar gastric contents volume and the PH value of gastric contents.There was no significant difference in serum hsCRP,fasting blood glucose and HOMA-IR between the two groups before operation.But on the first day,fasting blood glucose,HOMA-IR and hsCRP were significantly lower in the study group than in the control group(6.51 ±1.15 vs 7.49 ±0.57 mmol/L,P =0.038;4.34 ± 1.60 vs 6.09 ±2.81,P =0.043;40.45 ± 27.02 vs 80.02 ± 38.98 mg/L,P =0.03).Conclusion Preoperative oral administration of carbohydrate can obviously lower the postoperative blood glucose level and insulin resistance and alleviate postoperative inflammatory reaction.

Objective To investigate the correlation between the prognostic nutritional index (PNI) and clinicopathological features and long-term prognosis of gastric cancer patients after radical gastrectomy.Methods The clinical data of 135 gastric cancer patients who underwent radical gastrectomy in this hospital from 2002 to 2006 was analyzed retrospectively.The PNI value was calculated by serum albumin (g/L) + 5 x lymphocyte count (× 109/L).The receiver operating characteristic (ROC) curve and Youden index was used to determine the cutoff value of the PNI.Survival curves were described by the Kaplan-Meier method and compared by the Log-rank test.The univariate and multivariate analyses were performed with the Cox proportional hazard model to identify the prognostic factors.Result The mean PNI value was 47.3 ± 5.9.The mean values of the PNI in age (t =2.909,P =0.004),tumor size (t =2.227,P =0.028),tumor depth (t =3.314,P =0.001),negative lymph node (t =2.381,P =0.019),negative lymphovascular invasion (t =2.781,P =0.006) were significantly higher than those in patients without such factors.When the PNI was 47,the Youden index was maximal,with a sensitivity of 70％ and specificity of 63％.The mean age in high PNI group was significantly lower than that in low PNI group (x2 =6.443,P =0.011).Tumor infiltration depth in high PNI group was less than in low PNI group (x2 =7.394,P =0.007).The proportion of lymphovascular invasion in high PNI group was significantly lower than in low PNI group (x2 =4.540,P =0.033).The overall survival rate in high PNI group was higher than in low PNI group (P =0.002).The univariate and multivariate analyses showed that tumor location (OR,2.144 ; 95 ％ CI 1.239-3.712 ; P =0.006),lymph node metastasis (OR,4.887 ; 95 ％ CI 1.856-12.866 ; P =0.001),lymphovascular invasion (OR,1.842 ; 95％ CI 1.078-3.145 ; P =0.025) and the PNI value (OR,2.282 ; 95 ％ CI 1.344-3.874 ; P =0.002) were independent factors for predicting overall survival rate.Conclusions The PNI value is a simple and useful tool to predict the prognosis of patients with gastric cancer.

Objective To study the proliferation,collagen production and related gene expression in keloids and normal skin fibroblast.Methods Isolated primary cells of keloid fibroblasts (KFb,n=12) and normal human dermal fibroblasts (NFb,n=12) were identified,the cell viability and proliferating potential and the cell cycle were detected,and the difference on the collagen synthesis between KFb and NFb were compared.The expression of cell cycle-associated genes such as p21,p16,and p27 was dectected by real-time fluorescent quantitative PCR.Results The phase contrast optical microscopy imaging showed that both KFb isolated from keloid tissues and NFb from normal skin tissues possessed classic and similar fibroblast morphology.But there was a significant difference between cell proliferation,Hyp [(2.30±0.10) μg/ml vs.(1.66±0.13) μg/ml,P＜0.05] and collagen levels [(17.19±0.75) μg/ml vs.(12.37±0.94) μg/ml,P＜0.05].Compared with NFb,KFb exhibited more percentage of G2/M phase cells [(5.90±0.62)％ vs.(16.94 ％±1.93)％,P＜0.05]and less percentage of G0/G1 phase cells [(90.24 ±2.27)％ vs.(75.65±1.92)％,P＜0.05].Cell cycle related genes p16,p21 and p27 were low expressed.Collagen type Ⅰ was highly expressed at mRNA levels in KFb than that in NFb [0.84±0.11,1.32±0.2,1.69±0.12,4.33±0.27 in KFb vs.1.43±0.13,2.56±0.26,2.89±0.37,1.40±0.12 in NFb,P＜0.05].Conclusions There are cell dysfunction and abnormal cellular dynamics in keloid fibroblasts.The formation of keloid likely involves aberrant interactions of some genes that affected its development at different extents.

Objective Meningovascular syphilis is the intima inflammation of blood vessels caused by the syphilitic infec-tion, which is associated with the occurrence of ischemic stroke.The study analyzed the clinical, imaging features and prognosis for meningovascular syphilis so as to improve its diagnosis and treatment. Methods 14 patients diagnosed with meningovascular syphilis were collected prospectively from December 2007 to March 2015 in the neurological department of Jinling Hospital.The patients were followed for a period of 21.5(range 10.2~37.9)months,and the prognosis were evaluated. Results Patients with meningovascular syphilis presented with dizziness, hemiplegia, hemidysesthesia and cognitive decline.Lesions showed multiple, scattered on MR ima-ging, intracranial vascular stenosis was seen in the CTA/MRA, and the laboratory examination had characteristic changes.With a large dose and sufficient courses of penicillin treatment, meningovascular syphilis may hopefully get predominant effects.78.57%patients got good prognosis(modified Rankin Scale ,mRS≤2)at 3 months and 85.71% patients got goodlong-term prognosis(mRS≤2). Conclusion Meningovascular syphilis was usually presented as acute onset, lacks the specific clinical and neuroimaging manifesta-tions.Most patients has favourable prognosis after treatment of syphilis with full course of penicillin.

BACKGROUND: Peroxisome proliferation-activated receptor-gamma (PPARγ) is the member of nuclear receptor superfamily, and closely related with the formation of atherosclerosis.OBJECTIVE: To investigate the relationship between PPARγ C161→T gene polymorphism and coronary atherosclerotic heart disease (CAHD).DESIGN: Randomized controlled experiment SETTING: Department of Cardiology, Tonai Hospital of Huazhong University of Science and Technology; Department of Cardiology, Zhongnan Hospital of Wuhan University; Center for Human Genome Research,Huazhong University of Science and Technology; Department of Internal Medicine, Affiliated Hospital of Hubei University of Traditional Chinese Medicine; Institute of Geriatrics, Dongzhimen Hospital, Beijing University of Chinese Medicine PARTICIPANTS: Totally 203 CAHD patients aged (65±11) years, including 129 males and 74 females, were the inpatients and outpatients of Zhongnan Hospital of Wuhan University and Tonai Hospital of Huazhong University of Science and Technology from June 2002 to December 2005.And 156 cases of them were diagnosed by coronary arteriongraphy, among which 43 patients without coronary artery affection or with coronary stricture ＜ 50%, and 113 patients with coronary stricture ＞ 50 %. While 89 healthy physical examinees of Han race and mean (59±9) years old were enrolled as control group, including 56 males and 33 females. There was no blood relationship between controls and patients.METHODS: The experiment was conducted at Tongji Hospital of Huazhong University of Science and Technology from June 2002 to December 2005. PPARγ C161→T gene polymorphism was determined by polymerase chain reaction and restriction endonuclease fragment length polymorphisms. The radio-immunity technique, coronary angiography and clinical routine biochemical index were applied to analyze the genotypic frequency and allele frequency distributions as well as the relation between clinical data, biochemical index and different genotypes. The risk factors of CAHD were estimated in the patients of different genotypes.MAIN OUTCOME MEASURES: The genotypic frequency and allele frequency distributions, the relation between clinical data, biochemical index and different genotypes, along with the blood lipid, blood glucose, fasting insulin and body mass index (BMI).RESULTS: Totally 103 CAHD patients and 89 controls were involved in the result analysis of gene polymorphism and yielded different gene distribution frequencies.① In control group, "T" allele frequency was 0.213 and "C" allele frequency was 0.787, and in CAHD group, "T" allele frequency was 0.192 and "C" allele frequency was 0.808. There was no significant difference in the genotypic frequency and C, T allele frequencies between two groups (P ＞ 0.05).② The CC genotype was dominant in CAHD patients with coronary artery lesions, and showed significant differences from "T"allele carriers (CT+TT) (P ＜ 0.05). The CAHD risk in the "T" allele carries (OR: 0.56, 95% CI: 0.24-0.63) was much lower than that in the CC homozygote (OR: 1.92, 95% CI: 1.09-2.54).③ Apolipoprotein B in patients with CC genotype was obviously higher than that in patients with "T" allele (CT+TT) (P ＜ 0.05), and there was insignificant difference in the insulin resistance index (P ＞ 0.05).CONCLUSION: There is an important correlation between the substitution of PPARγ C161→T and CAHD, and "T" allele carriers demonstrate a lower risk of CAHD.

The genome of CK/CH/SD09/005, an isolate of infectious bronchitis virus (IBV), was characterized to enable the further understanding of the epidemiology and evolution of IBV in China. Twenty-five pairs of primers were designed to amplify the full-length genome of CK/CH/SD09/005. The nucleotide sequence of CK/CH/SD09/005 was compared with reference IBV strains retrieved from GenBank. The phylogenic relationship between CK/CH/SD09/005 and the reference strains was analyzed based on S1 gene sequences. The complete genome of CK/CH/SD09/005 consisted of 27691 nucleotides (nt), excluding the 5' cap and 3' poly A tail. The whole-genome of CK/CH/SD09/005 shared 97 - 99% nucleotide sequence homology with the GX-NN09032 strain, which was the only complete genome that was closely related to CK/CH/SD09/005. When compared with all reference strains except GX-NN09032, CK/CH/SD09/005 showed the highest similarity to ck/CH/LDL/091022 and SDIB821/2012 (QX-like) in the replicase gene (Gene 1) and 3'UTR, with a sequence identity rate of 97% and 98%, respectively. However, CK/CH/SD09/005 exhibited lower levels of similarity with ck/CH/LDL/091022 and SDIB821/2012 in S-3a-3b-3c/ E-M-5a-5b-N with a sequence identity of 72% - 90%. CK/CH/SD09/005 showed the highest level of nucleotide identity with Korean strain 1011, and Chinese strains CK/CH/LXJ/02I, DK/CH/HN/ZZ2004 and YX10, in ORF 3c/E (97%), 5a (96%), 5b (99%) and N (96%), respectively. ORFs 3a, 3b and M of CK/CH/SD09/005 exhibited no more than 90% homology with the reference strains, excluding GX-NN09032. The phylogenic analysis based on the S1 gene revealed that CK/CH/SD09/005 and 39 published strains were classified into seven clades (genotypes). CK/CH/SD09/005 was distributed in clade IV with several isolates collected between 2007 and 2012. CK/CH/SD09/005 showed 66% - 69% and 72% - 81% nucleotide identities with the IBV strains of other six clades in the S1 and S2 subunits, respectively. More over, multiple substitutions were found throughout the entire S gene of CK/CH/SD09/005, while insertions and deletions were located within the S1 gene. These results indicated that CK/CH/SD09/005 is a novel variant that may be derived from the QX-like strains that are prevalent in China. Multiple genetic mechanisms, including recombinations, mutations, insertions and deletions, are likely to have contributed to the emergence of this IBV strain.
Animals ; Chickens ; China ; Coronavirus Infections ; veterinary ; virology ; Genome, Viral ; Genomics ; Infectious bronchitis virus ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; virology ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics

As medicine and food plants, the market demand of Zamthoxyli Pericarpium is extensive, and the sanshool in Zamthoxyli Pericarpium is getting attention gradually. However, because of the low content and not establishing rating evaluation method for sanshool and lack of standards, the research on substance basis and physiological activity is necessary. This article reviewed the extraction, separation, purification and biological activities of sanshoolin Zamthoxyli Pericarpium in recent years to provide development and utilization of Zamthoxyli Pericarpium.