Streptococcus pneumoniae(S.pn) is an opportunity pathogenic bacteria,environmental factors play a key role in the pathogenicity of S.pn. It is important to study virulent gene in vivo. The S.pn suicide plasmid containing gfp reporter was constructed by fusing the genes pneumolysin and gfp,in which gfp is an excellent molecule probe in vivo. The plasmid was integrated to No.22 S.pn by homologous recombination. The recombinant S.pn was gained and evaluated in aspects of fluorescence excitation, biological character and physio-activity. The results showed it is efficient and available to report the expression of virulent genes in vivo and in vitro, which will provide a new easy method for evaluating and screening the virulent genes of S.pn in vivo.

OBJECTIVETo investigate and research the appraisal scores of the symptoms and physical signs index for the evaluation of the clinical efficacy of acupuncture and manipulative regulation of knee muscle balance for the treatment of knee osteoarthritis, and provide clinical basis for the treatment.

METHODSFrom January 2008 to December 2009, 121 patients with knee osteoarthritis were randomly divided into two groups. In the treatment group there were 63 cases of 83 knees including 16 males and 47 females, with an average age of (59.88 +/- 7.97) years; in the control group there were 58 cases of 73 knees including 13 males and 45 females, with an average age of (57.95 +/- 10.37) years. The patients in the treatment group were treated with acupuncture plus manipulative regulation of knee muscle balance, and the patients in the control group were treated with Diclofenac Sodium Sustained Release Tablets. The appraisal scores of the symptoms and physical signs index, numerical rating scale of pain, joint function, joint swelling were evaluated before and after the treatment, as well as 3 months after the treatment. All data were statistical analyzed by package SPSS 10.0.

RESULTS1) In the treatment group, before and after treatment the appraisal scores of the symptoms and physical signs index were (39.81 +/- 3.92) and (9.69 +/- 8.08); numerical rating scale of pain were (7.61 +/- 0.97) and (2.17 +/- 2.09); joint function were (1.47 +/- 0.50) and (0.61 +/- 0.58); joint swelling were (1.23 +/- 0.79) and (0.42 +/- 0.52). As well in the control group, above data were (39.89 +/- 3.78), (13.62 +/- 7.83), (7.55 +/- 0.71), (3.34 +/- 2.32), (1.33 +/- 0.47), (0.93 +/- 0.67), an (0.97 +/- 0.88), (0.58 +/- 0.52) respectively. Both group had obvious differences in the appraisal scores of the symptoms and physical signs index, numerical rating scale of pain, joint function, joint swelling between after and before treatment. The comparison between the two groups suggested that after the treatments the treatment group had better therapy effect than those of the control group in the appraisal scores of the symptoms and physical signs index, numerical rating scale of pain, joint function, but both group had no differences in the appraisal scores of joint swelling. 2) After the treatment, 12 patients in the treatment group were clinically cured, 48 had remarkable effects,19 good, and 4 ineffective. As well in the control group, above data were 5, 31, 34 and 3 respectively. Three months after the treatment, above data were 8, 42, 27 and 6 respectively in the treatment group. In the control group, above data were 5, 21, 37 and 10 respectively. The treatment group was superior to the control group after the treatments and 3 months after the treatment.

CONCLUSIONAcupuncture plus manipulative regulation of knee muscle balance can effectively improve the clinical symptoms and knee joint's motor function of patients with knee osteoarthritis, and can avoid the further development of disease. It is a proved effective method for knee osteoarthritis.

Acupuncture Therapy ; Aged ; Combined Modality Therapy ; Female ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Osteoarthritis, Knee ; physiopathology ; therapy

OBJECTIVETo localize susceptibility loci in Chinese systemic lupus erythematosus (SLE) cohort by linkage disequilibrium mapping the genomic interval in human chromosome 16 so as to understand whether the pathogenesis of human SLE is related to chromosome 16.

METHODSFive microsatellite markers in chromosome 16 spanning from 57.79 cM to 65.1 cM were used for fluorescence based genotyping in 157 SLE families. Evidence for linkage disequilibrium was assessed using the extended transmission disequilibrium test (ETDT) program and Genehunter software. Susceptibility gene was searched in the positive locus, and real-time PCR method was used to detect gene expression.

RESULTSWith the ETDT, evidence for linkage disequilibrium of the marker D16S409 (P=0.0278) and D16S517 (P<0.0001) with SLE were found. It was observed that 271 bp allele of D16S517 was much in evidence for preferential transmission, while 277 bp allele was found to be transmitted less often than expected. Genehunter analysis is concordant with ETDT. By real-time PCR, OAZ(OLF1/ EBF-associated zinc finger protein) gene which is located in the positive locus showed higher expression in SLE patients than in normal controls.

CONCLUSIONResults reveal that 16q12 (58.46 cM) is associated with Chinese SLE. OAZ is a likely susceptibility gene in this locus for SLE.

Adolescent ; Adult ; Alleles ; China ; Chromosomes, Human, Pair 16 ; genetics ; Cohort Studies ; DNA ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Humans ; Linkage Disequilibrium ; Lupus Erythematosus, Systemic ; genetics ; Male ; Microsatellite Repeats ; Middle Aged

OBJECTIVETo discuss the urine biomarkers in 1,3-butadiene exposed workers, and to provide basement for establishing biological limit value.

METHODS44 BD exposed workers as exposure group and 25 BD non-exposed people as control group including 12 workers in boiler workshop in the same factory and 13 people in one public institute, we collected their in-end-of shift urine, then detected urine BD-derived mercapturic metabolites [3,4-dihydroxybutyl mercapturic acid (DHBMA),1- and 2-monohydroxy-3-butenyl mercapturic acid (MHBMA)] concentrations using UPLC-MS/MS method. Meanwhile, we detected air BD concentration with GC-FID in the workplace, and compared their relationship.

RESULTSlgDHBMA and lg (MHBMA + DHBMA) levels in exposed group (lgDHBMA: 2.51 ± 0.44) µg/L, lg [MHBMA + DHBMA: (2.68 ± 0.27) µg/L] were higher than which in control group (lgDHBMA: (2.20 ± 0.25) µg/L, lg(MHBMA + DHBMA: (2.49 ± 0.34) µg/L), and the differences were significant (P < 0.01). Urine DHBMA was obviously influenced by air BD concentrations (r = 0.539, P = 0.001). The equation of Multiple Regression Analysis was y = 2.417 + 0.520x (x represents air BD dose, and represents urinary DHBMA level). Adjusted R(2) of this model was 0.262. Urinary MHBMA was not affected by smoking, alcohol and years of works.

CONCLUSIONUrine metabolite DHBMA in BD-exposed workers might be major biological exposure indice.

Adult ; Biomarkers ; urine ; Butadienes ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; adverse effects ; Surveys and Questionnaires

Adult ; External Fixators ; Female ; Humans ; Male ; Middle Aged ; Recovery of Function ; Tibia ; anatomy & histology ; pathology ; physiopathology ; Tibial Fractures ; diagnosis ; diagnostic imaging ; physiopathology ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome

OBJECTIVETo investigate the inhibitory effect of compound cantharides capsules on the proliferation of xenografts of human hepatocellular carcinoma HepG(2215) in mice and their mechanism of action.

METHODSOne hundred healthy Balb/c mice (5-week old, male:female 1:1) were used in this study. Mouse models of human HepG(2215) hepatocarcinoma were established. The tumor-bearing mice were divided into five groups randomly. The control group A received daily intragastric administration of physiologic saline. The intervention groups B1, B2 and B3 were treated with compound cantharides capsule in a dose of 12.5 mg×kg(-1)×d(-1), 25 mg×kg(-1)×d(-1) and 37.5 mg×kg(-1)×d(-1), respectively, for 10 consecutive days. The group C had intraperitoneal injection of cyclophosphamide (25 mg×kg(-1)×d(-1)) for 10 consecutive days. The mice were sacrificed after the completion of administration. The tumors were taken out, the tumor volume was measured, the inhibitory rate of body weight was calculated, and the serum AFP concentration and the level of HBV DNA were determined. The survival of each group mice was analyzed. The levels of mRNA expression of apoptosis-related genes were assayed by quantitative RT-PCR. Apoptosis in the tumor cells was assayed with TUNEL staining. Flow cytometry was used to detect the levels of CD3(+), CD19(+), CD4(+) and CD8(+), and microvessel density (MVD) of the tumors was assessed by immunohistochemistry.

RESULTSAfter completion of the treatment, the inhibition rate of tumor growth of the groups B1, B2 and B3 was 29.8%, 38.7% and 48.1%, respectively, and that of the group C was 52.4%, with a significant difference among the groups (P < 0.05). The median survival time of the groups A, B1, B2, B3 and C was (30.0 ± 3.2) days, (49.0 ± 5.1) days, (50.0 ± 5.2) days, (57.5 ± 6.5) days and (49.0 ± 4.7) days, respectively. The median survival time of the group B3 was significantly longer than that of other groups (P < 0.05). The serum AFP level in the groups A, B1, B2, B3 and C was (492.7 ± 48.5) ng/ml, (281.2 ± 25.6) ng/ml, (194.3 ± 18.7) ng/ml, (170.1 ± 15.8) ng/ml and (138.7 ± 12.5) ng/ml, respectively, indicating that it was significantly inhibited in the group C. The inhibition rate of HBV DNA replication of the groups B1, B2, B3 and C was (46.0 ± 5.1)%, (65.5 ± 6.9)%, (81.3 ± 7.8)% and (19.5 ± 2.1)%, respectively, showing that compound cantharides capsules inhibited HBV DNA replication in a dose-dependent manner. The apoptosis rate of the groups A, B1, B2, B3 and C was (0.27 ± 0.03)%, (7.18 ± 2.12)%, (9.17 ± 2.42)%, (11.27 ± 3.03)% and (5.44 ± 2.45)%, respectively, and that of the group B3 was significantly higher than that of the groups A, B1, B2 and C (P < 0.05). The expression level of bax mRNA was significantly higher than that of the group C (P < 0.05). The drug could significantly decrease the bcl-2 mRNA expression level, more remarkably along with the increasing dose of cantharides, and it was significantly lower than that in the group C (P < 0.05). The levels of CD4(+), CD8(+), CD3(+) and CD19(+) were significantly higher than that in the groups A and C (P < 0.05). The value of MVD of the group B3 was significantly lower that that of groups A and C (P < 0.05).

CONCLUSIONCompound cantharides capsules may inhibit the replication of HBV DNA in HepG(2215) cells, inducing apoptosis in the tumor cells, enhancing the immune function to inhibit the growth of liver cancer cells in mice, and significantly prolong the median survival time of tumor-bearing mice.

Animals ; Antineoplastic Agents ; administration & dosage ; pharmacology ; Antiviral Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Cantharidin ; administration & dosage ; pharmacology ; Capsules ; DNA Replication ; DNA, Viral ; Drug Combinations ; Female ; Hep G2 Cells ; Hepatitis B virus ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Microvessels ; pathology ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Tumor Burden ; drug effects ; Virus Replication ; Xenograft Model Antitumor Assays ; alpha-Fetoproteins ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

Objective To design,synthesize and screen high efficient small interfering RNA(siRNA) targeting to cyclooxygenase-2(COX-2)on rheumatoid arthritis synovial fibroblasts(RASF).To further study the effect of specific COX-2 siRNA interfering on mediators of inflammatory cytokines.Methods Four pairs of siRNA for human COX-2 mRNA were synthesized by utilizing RNA design software,while another random sequence was designed as control.They were divided into group A to H.Among them,group A was used as the negative control(CTL),and group B to F were transfected as random siRNA(NC),1#～4#siRNA in order. These siRNAs were transferred into RASF by LipofectAMINE2000 package and PMA(phorbol-12-myristate- 13-acetate)was added into each culture and with a final concentration of 100 nmol/l.RASF was collected 48 hours after transfection.The expression of hCOX-2 at mRNA level was determined by reverse transcription- polymerase chain reaction(RT-PCR)and hCOX-2 protein level by Western Blot.The supernatant levels of PGE_2,IL-1?,IL-6,TNF-?and vascular endothelial growth factor(VEGF)of the above groups were detected by ELISA.Results The levels of hCOX mRNA and protein in RASF treated with 4-#siRNA were significantly lower than those of the negative control and other groups.The level of PGE_2 and cytokines like IL-1?,IL-6, TNF-?and VEGF in the supernatant were lower in the 4#siRNA group than in other groups.Conclusion 4#siRNA can effectively inhibit the expression of COX-2 mRNA and the synthesis of the COX-2 protein in human synovial fibroblasts.The level of PGE_2,IL-1?,IL-6,TNF-?and VEGF is the lowest in the super- natant.Thus 4#siRNA has been confirmed to specifically block the COX-2 in human synovial fibroblasts.

OBJECTIVETo study the feasibility and clinical efficacy of a minimally invasive sinus tarsi approach in the treatment of Sanders II calcaneus fractures.

METHODSFrom August of 2015 to July of 2016, 13 patients(totally 13 feet) with Sanders II intra-articular calcaneus fractures were treated via the minimally invasive sinus tarsi approach. The Böhler angle, Gissane angle and the length, width and height of calcaneus were compared between pre-operation and post-operation. The AOFAS ankle and foot scoring system of the orthopaedic ankle foot Association was used to evaluate the efficacy.

RESULTSAll the patients were followed up, and the duration ranged from 6 to 15 months, with an average of 9.5 months. No incision complications occurred. The Böhler angle was increased from preoperative (18.82±5.11)° to postoperative(26.63±4.45)°(=-4.16,=0.000). The Gissane angle was increased from preoperative(111.07±15.36)° to postoperative (124.56±8.71)° (=-2.75,=0.011). The length, width, height of calcaneus were absolutely improved from preoperative(69.82±5.95) mm, (42.07±3.68) mm, (41.20±3.90) mm to preoperatively(72.61±5.46) mm, (39.10±4.02) mm, (44.03±3.33) mm. According to the AOFAS, 8 patients got an excellent result, 4 good and 1 poor, and the postoperative mean score was 88.2±5.9.

CONCLUSIONSThe limited open sinus tarsi approach could be used successfully to treat displaced Sanders II fractures with less injury and effectively restored the surface of subtalar joint, however the method is not fit for the patients with comminuted fracture in lateral wall and great change in the length, width, height, varus and valgus of calcaneus.

OBJECTIVETo identify epidemic status of murine typhus in Hongta areas of Yuxi city and to provide evidence for control and prevention of the disease.

METHODSSerologic survey was conducted among residents and rodents. Isolation of Rickettsia moseri was performed.

RESULTSThe overall infection rate among general population was 28.92% (96/332) with geometric meantiter (GMT) as 10.83 and there was no difference between males and females (26.71%, 43/161 vs. 30.99%, 53/171, P > 0.05). Significant differences were found between age groups (P < 0.05) with positive rates of 29.63% (8/27), 18.06% (13/72), 39.62% (42/106), 27.50% (22/80) and 23.40% (11/47) among age groups 0-6, 7-18, 19-39, 40-59 and over 60, respectively. The overall rate of infection in mouse was 44.95% (89/198) with GMT as 30.30. Five isolates of R. moseri from mouse specimen, three from fleas plus one case of murine typhus were diagnosed. Rattus norvegicus and Rattus flavipectus were the predominant species of rodent animals (99.49%, 197/198) and Xenopsylla cheopis was the major species of vector (74.26%, 303/408). Flea index and mouse density were 2.06 and 11.13% respectively.

CONCLUSIONHigh infection rates on R. moseri were demonstrated in rodents and residents as well as high risk of murine typhus outbreak might occur in these areas.

Adolescent ; Adult ; Animals ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Mice ; Middle Aged ; Rats ; Rodent Diseases ; epidemiology ; microbiology ; transmission ; Siphonaptera ; microbiology ; Typhus, Endemic Flea-Borne ; epidemiology ; microbiology ; transmission ; Young Adult

BACKGROUNDWe hypothesize that increased atrial oxidative stress and inflammation may play an important role in atrial nerve sprouting and heterogeneous sympathetic hyperinnervation during atrial fibrillation (AF). To test the hypothesis, we examined whether the antioxidant and anti-inflammatory treatment with probucol attenuates atrial autonomic remodeling in a canine model of AF produced by prolonged rapid right atrial pacing.

METHODSTwenty-one dogs were divided into a sham-operated group, a control group and a probucol group. Dogs in the control group and probucol group underwent right atrial pacing at 400 beats per minute for 6 weeks, and those in the probucol group received probucol 1 week before rapid atrial pacing until pacing stopped. After 6-week rapid atrial pacing, general properties including left atrial structure and function, atrial hemodynamics and the inducibility and duration of AF were measured in all the groups. Atrial oxidative stress markers and serum C-reactive protein (CRP) concentration were estimated. The degree of nerve sprouting and sympathetic innervation at the right atrial anterior wall (RAAW) and the left atrial anterior wall (LAAW) were quantified by immunohistochemistry, atrial norepinephrine contents were also detected. Atrial beta-nerve growth factor (beta-NGF) mRNA and protein expression at the RAAW and LAAW were assessed by real-time quantitative RT-PCR and Western blotting respectively.

RESULTSAtrial tachypacing induced significant nerve sprouting and heterogeneous sympathetic hyperinnervation, and the magnitude of nerve sprouting and hyperinnervation was higher in the RAAW than in the LAAW. Atrial beta-NGF mRNA and protein levels were significantly increased at the RAAW and LAAW, and the upregulation of beta-NGF expression was greater at the RAAW than at the LAAW in the control group. The beta-NGF protein level was positively correlated with the density of sympathetic nerves in all groups. Probucol decreased the increase of CRP concentration and attenuated atrial oxidative stress caused by atrial tachypacing. In addition, probucol could effectively inhibit atrial beta-NGF upregulation, significantly attenuate atrial nerve sprouting and heterogeneous sympathetic hyperinnervation, and dramatically reduce the inducibility and duration of AF.

CONCLUSIONSThe atrial over-expression of beta-NGF possibly caused by increased oxidative stress and inflammation may be the main mechanism underlying atrial autonomic remodeling during AF. Probucol attenuates atrial autonomic remodeling possibly by its antioxidant and anti-inflammatory actions.

Animals ; Antioxidants ; therapeutic use ; Atrial Fibrillation ; drug therapy ; Blotting, Western ; C-Reactive Protein ; metabolism ; Cardiac Pacing, Artificial ; adverse effects ; Disease Models, Animal ; Dogs ; Electrocardiography ; Female ; Heart Atria ; Immunohistochemistry ; Male ; Nerve Growth Factor ; genetics ; metabolism ; Norepinephrine ; metabolism ; Probucol ; therapeutic use ; Reverse Transcriptase Polymerase Chain Reaction