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 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

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

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.

Objective:To investigate the expression pattern and prognostic value of NUF2(Ndc80 kinetochore complex component)in breast cancer(BC).Methods:In the present study,the trancriptional level and survival information of NUF2 was analyzed by ONCOMINE dataset,bc-GenExMiner,Kaplan-Meier Plotter and TCGA.Results:NUF2 was highly expressed in BC com-pared with normal samples(P＜0.05).In addition,the BC patients younger than 51 years old(P＜0.0001),nodal metastasis(P=0.0168),with basal-like BC(P＜0.0001),orestrogen receptor(ER)-negative(P＜0.0001),progesterone receptor(PR)-negative(P＜0.0001),human epidermal growth factor receptor-2(HER2)-positive(P＜0.0001)and triple-negative BC(P＜0.0001)pathological clas-sification have a higher transcriptional level of NUF2.Survival analysis indicated that patients with low expression level of NUF2 have a better relapse-free survival(RFS,HR=1.63,P＜0.0001).Fur-ther anaysis revealed that the NUF2 expression was negatively related to the RFS of BC patients with ER-positive or PR-positive or HER2-negative pathological classification(HR=2.40,P=0.0015).Conclusion:NUF2 play an important role in BC and can be a potential prognostic bio-marker for ER-positive or PR-positive or HER2-negative BC patients.

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.

Objective To investigate the indications and complications of the total thyroidectomy in the management of thyroid cancer. Methods Fifty-one patients with thyroid cancer operated with total thyroidectomy in a period from April 2007 to June 2010, were retrospective analyzed. There were 48 cases of papillary adnocarcinoma, 1 follicular adnocarcinoma, and 2 medullary carcinoma as well, 2 cases with hyperthyroidisms, 3 cases with remote metastasis. There were 17 males and 34 females who in their age ranged from 5 years old to 82 years old with a middle age of 50 years old. Among the 45 cases who were initial surgical management patients, there were 17 T1, 13 T2, 7 T3, 8 T4; and there were 31 NO, 11 N1a,3 N1b, 2 M1. The recurrent laryngeal nerve and the parathyroid gland, and supper laryngeal nerve were exposed routinely. Lymph nodes in Level Ⅵ were dissected in all patients, and Level Ⅱ -Ⅳ dissected in 25patients. 131Ⅰ were administrated in 8 patients. Results Patients were followed up from 1 month to 36months. There were no patients died and lost of follow up. The two years survival rate was 100. 0% ( 15/15 )Pathological examination showed that the multiple focal disease rate was 42. 2% ( 19/45 ), the cervical lymph nodes metastases rate was 51.0% (26/51). There was no permanent paralysis of recurrent laryngeal nerve and outer branch of supper laryngeal nerve. The permanent hypoparathyroidism rate was 3. 9%(2/51). One case with contralateral lymph node metastasis was reoperated and survived without disease.Two cases with remote metastasis were alive with steadied disease. Conclusions Under the skilled hand,total thyroidectomy is a safe procedure in the management of thyroid cancer. Totally exposing the parathyroid gland and laryngeal nerve is the key point to prevent the major complications.