0.05). Conclusion Jingu Tongxiao Granules could effectively relieve the pain, morning stiffness and motor impairment of knee joint osteoarthritis safely. Compared with the original form, the efficacy and safety of Jingu Tongxiao Granules were the same.

BACKGROUND: Hanshibi granule indicates rheumatism due to blockage of cold and damp.OBJECTIVE: To observe the effects and safety of intervention of hanshibi granule on rheumatism due to blockage of cold and damp and the comparative observation was carried on by taking zhengqingfengtongning as the control drug.DESIGN: Case randomized-controlled observation was designed.SETTING: Luoyang Bone-setting Hospital of Henan Province.PARTICIPANTS: Totally 400 cases of rheumatism due to blockage of cold and damp visited in Luoyang Orthopedic Hospital of Henan Province were volunteered in the observation from May 2000 to February 2002.Hanshibi granule was composed of fuzhi (Radix Aconiti Praeparata),huangqi (Radix Astragali), weilingxian (Radix Clematidis), more than 10 herbs, 10 g each bag. Zhengqing fengtongning tablet is 20 mg each tablet.METHODS: Based on the randomization and sequence of visiting, the cases were divided into observing group and the control at the ratio of 3:1.In the observing group, hanshibi granule was administrated, 1 bag/time,3 times/day, infused with water. In the control, zhengqing fengtongning tablet was administered orally, 2 tablets/time, 3 times/day; 1 week later,if there was no harmful reaction, it was administered 3 tablets/time,3 times/day. For rheumatic arthritis (RA) and ankylosing spondylitis (AS),the drugs were taken for 2 months as 1 course, and for knee osteoarthritis,the drugs were taken for 1 month as 1 course. The changes of ESR, C-reactive protein (CRP) and rheumatic factor (RF) and uric acid (UA) were determined and recorded. Criteria on evaluation: ① Evaluation on therapeutic effects on syndromes: Remarkable effect: the symptoms were improved remarkably after treatment and symptom integral was 0-1 score or decreased by ≥ 2/3. Effect: the symptoms were improved after treatment and the symptom integral was decreased by≥1/3.② Evaluation on therapeutic effects on symptoms: Remarkable effect: The symptom integral was reduced to 0 score or reduced by 2 scores after treatment, such as joint pain, swelling, tender pain, morning stiffness and functional impairment. Effect: the integral was reduced by 1 score after treatment. ③ Evaluation on total therapeutic effects on diseases: Remarkable effect: the symptoms and physical signs disappeared basically, the main laboratory indexes were improved remarkably and reduced by ≥ 50%. Effect: The symptoms and physical signs were alleviated and the main laboratory indexes were improved.MAIN OUTCOME MEASURES:① Total clinical therapeutic effect,therapeutic effect on different symptoms and diseases between two groups.②Safety of hanshibi granule.RESULTS: Totally 400 cases were employed, of which, 300 cases were in observing group and 100 cases in the control. In observing group, 19 cases were dropped out and 281 cases were in the statistics. In the control,6 cases were dropped out and 94 cases were in the statistics.①Comparison of total clinical therapeutic effects between two groups: The total effective rate in the observing group was higher significantly than that in the control (94.31％, 84.04％, P ＜ 0.01). ② Comparison of therapeutic effects on different symptoms between two groups: There was no significant difference in the therapeutic effects on joint pain, swelling,tender pain, functional disturbance and morning stiffness between two groups (P ＞ 0.05). ③ Comparison of therapeutic effects on different diseases between two groups: There was no significant difference in the therapeutic effects on RA, AS and knee osteoarthritis (P ＞ 0.05). ④ Safetyof hanshibi granule: abnormal urine routine for a part of cases before treatment was improved after treatment. ST-T wave alternation in ECG for a part of cases before treatment was not aggravated obviously after medication.CONCLUSION: Both hanshibi granule and zhengqingfengtong tablet, the control drug improves significantly joint pain, swelling, tender pain, functional impairment and morning stiffness in rheumatism due to blockage of cold and damp, in which, the effect of intervention of hanshibi granule is superior to that of the control drug, without obvious harmful reactions;therefore, such drug can be taken as a safe and effective one for rheumatism due to blockage of cold and damp.

BACKGROUND: The biomechanical property(BP) of the bone is decided by its geometric structure and component material. Merely pursuing increase of the bone mineral density(BMD) might lead to deterioration of bone BP.However at present, some researohes on therapeutic action on osteoporosis emphasize excessively medical influence to BMD, and the change in the holistic BP of the bone in osteoporotic zone and its mechanism still need to investigate deeply.OBJECTIVE: To probe into the action and its mechanism of "the kidney tonifying compound of the Traditional Chinese Medical (TCM) "on BP of cortical bone in ovariectomized osteoporotic model rats.DESIGN: Completely randomized controlled experiment based on experimental animals.SETTING: Laboratory of Biomedical Engineering, Luoyang Hospital and Institute of Traditional Chinese Orthopedics and Traumatology in Henan Province.MATERIALS: The experiment was completed from November 2000 to July 2001 at Research Laboratory of Biomedical Engineering,Luoyang Institute of Traditional Chinese Orthopedics and Traumatology of Henan Province. The healthy Wistar female rats aged 10 months,weighing(350±20) g.METHODS: Fifty Whistar female rats aged 10 months were randomly divided into 5 groups: the normal, model, premarin-treated, xianling gubao-treated and migu capsule-treated with 10 in each group. The normal group was only given sham operation and the other four groups were ovariectomized. The rats after operation were fed normally for ninety days.Since the 91st day after operation,the rats had been given the medicines for 90 days and then killed. The thighbones were taken out,then BMD,femoral geometry sizes and BP were determined.MAIN OUTCOME MEASURES: ① The primary sequel was the comparison of the parameters of femoral BP. ② The secondary sequel was the changes in parameters of femoral geometric structure, area of cortical bone and BMD of every midsectional fomur.RESULTS: Femoral BP worsened significantly,its mechanical intensity reduced,its external diameter diminished,cortical bone area decreased and femoral BMD lowered in osteoporotic model rats. In comparison with the above,in "the kidney tonifying compound ofTCM " groups(migu capsule group and xianling gubao group) femoral BP raised significantly, its mechanical intensity advanced,its external diameter augmented,cortical bone area aggrandized and femoral BMD enhanced.CONCLUSION: "The kidney tonifying compound of TCM" can improve BP of the cortical bone(thighbone) in ovariectomized osteoporosis rats. Its primary mechanism of action is that the TCM compound prescription could enhance"the mechanism of biomechanical response and regulation"(MBRR) of macrostructure of cortical bone,consequently increase femoral external diameter,aggrandize cortical bone area and enhance BMD in ovariectomized rats.

Objective To obtain shRNA sequences that can stably block the expression of Nuclear Factor kappa- B (p65) in the prostate cancer cell line LNCaP and construct the lentivirus vector.And validate the gene function of p65 in the cell line. Methods According to p65 genetic information, we design siRNA1, siRNA2, siRNA3 those three siRNA sequences targeting the ods area of p65 gene and then form the corresponding four pairs of complementary single strand DNA of shRNA, including the sense strand and the antisense strand. The synthetic shRNA sequence was inserted into the empty pSIH1-H1-copGFP shRNA Vector, and after transfecting the prostate cancer cells , the inhibitory effect of p65 mRNA by different sequences was detected through real-time PCR, and the inhibitory effect of p65 protein expression was detected by Western-blotting. Thus we can obtain highly effective shRNA sequences in the inhibition of p65 in prostate cancer cells. MTT, flow cytometry, transwell were chosen to test the cell growth, migration and invasive power in vitro to compare the difference of the experimental group, control group and negative group. Results The third shRNA sequence had the best inhibitory effect and the inhibitory effect of p65 mRNA in prostate cancer cell line was 59 % and the protein was 81%. It's position locates in p65 (NM_021975 ) 1096-1113 and it's stemloop sequence is 5'-GATCCGCCCTATCCCTTTACGTCATTCAAGAGATGACGTAAAGGGATAGGGCTTTTTG-3'. After transfecting, the prostate cancer cell line had the low expression of p65 stably. Through MTT, we got the growth curve, which showed that the growth ability of experimental group was significantly decreased compared with the control group and the Logarithmic growth didn't appear in the first 96 hours. Flow cytometry test displayed that the percentage of G0-G1-phase cells in experimental group was 61.49%, and the control group was 44.89%, idle group was 41.52%, which was increasing oberviously. The S-phase cells in the experimental group was 28.58%, compared with the 47.36% and 46. 10% diminished. The results of transwell showed that the experimental group had 16. 5000±6. 62076 cells and the other two groups had 45. 6333 13. 54159 and 36. 8333±5. 68412 cells, which showed the invasive power of experimental group was significantly declined(P＜0.05).Conclusions It's successful to obtain shRNA sequences that can stably block the expression of p65 in the prostate cancer cell line LNCaP and construct the lentivirus vector. p65 can positively regulates the biological behavior of prostate cancer LNCaP cell line in the cell growth, migration and invasive power.

Objective To investigate the capsular polysaccharide (CPS) serotypes and molecular characteristics of carbapenem resistant hypermucoviscous Klebsiella pneumoniae (CR-HMKP) and study the possible mechanism of carbapenem resistance. Methods A retrospective study was conducted on 18 nonduplicate CR-HMKP strains which were collected from the First Affiliated Hospital of Nanchang University from 2012 to 2016. The clinical data were retrieved from medical records. The capsular serotypes, resistance genes and virulence factors were detected by polymerase chain reaction and DNA sequencing. Antimicrobial susceptibility testing was determined on VITEK 2 compact system. The CR-HMKP strains were characterized molecularly by using PCR, multilocus sequence typing (MLST) and pulsed field gel electrophoresis (PFGE). Modified carbapenem inactivation method was used to screen carbapenemase-producing strains. Plasmid conjugation transfer experiments were carried out to study transmission of carbapenem resistance. Results Eighteen (2.7％) CR-HMKP isolates were identified, which belonged to 4 serotypes, including wzi128-K1 (n=1), wzi206-K57 (n=1), wzi2-K2 (n=2), and wzi64-K14.64 (n=14). PCR and sequencing analysis identified blaNDM-1 gene in 2 CR-HMKP strains, blaKPC-2 gene in 17 strains, qnrS1 gene in 18 strains, blaCTX-M-3 gene in 3 strains, blaCTX-M-14 gene in 18 strains, blaTEM-1 gene in 16 strains, blaSHV-12 gene in 17 strains, and rmtB in 5 strains. All the 18 CR-HMKP strains carried virulence-associated genes, including rmpA (88.9％, 16/18), magA (5.6％, 1/18), iroN (83.3％, 15/18), aerobactin (27.8％, 5/18), rmpA2 (66.7％, 12/18) and mrkD (100％, 18/18). Three sequence types (STs) were identified by MLST, including ST11 (15 strains), ST86 (2 strains), and ST412 (1 strain). PFGE resulted in three major PFGE clusters, of which cluster A corresponds to ST1 isolates, and cluster B corresponds to ST86 isolates, and cluster C corresponds to ST412 isolates. All the blaKPC-2- positive strains belonged to ST11. Plasmid conjugation was successful in 5 (27.8％) of the 18 CR-HMKP isolates. Conclusions wzi64-K14.64 is the predominant capsule serotype of the CR-HMKP strains in this hospital. KPC-2 gene conjugationmay contribute to the emergence of CR-HMKP isolates. In addition, CRHMKP strain may be the highly prevalent ST11, and highly virulent CPS serotypes harboring K1/K2.

Objective To investigate the antimicrobial resistant mechanisms of high level carbapenem resistant Klebsiella pneumoniae infection of burn patients .Methods A retrospective study was conducted on totally 18 non-repetitive high level CR-KP which were isolated from burn patients hospitalized between July 2014 and June 2015.MIC of antibiotics were determined by using the GN 13 cards and agar dilution method.The specific PCR and DNA sequence analysis were performed to confirm the β-lactamase type.Plasmid conjugation transfer experiments and southem hybridization were applied to study the mode of carbapenem resistance transmission .Outer membrane proteins ( Omps) were isolated and examined by PCR and ( sodium dodecyl sulfate polyacrylamide gel electrophores ) SDS-PAGE.Pulsed-field gel electrophoresis( PFGE ) and Multilocus sequence typing ( MLST ) was used to determine the genotypes . Results Susceptibility of antimicrobial agents indicated that all these strains with multiple drug resistance . The resistance rate to piperacillin/tazobactam, ceftriaxone, levofloxacin, ciprofloxacin, gentamicin, tobramycin, cefotetan, ceftazidime, cefepime, aztreonam, imipenem, and meropenem was 100％ (18/18).Moreover, the resistance rate of CR-KP isolates to amikacin was 72.2％ ( 13/18 ) , compound sulfamethoxazole was 61.1％(11/18), tigecycline was 0％(0/18).Conjugation study with Escherictda coli J53 resulted in the transfer of significant reduced carbapenem susceptibility from donors (MICs increased at least 8-fold).By PCR, eighteen strains of Klebsiella pneumoniae carried NDM-1 gene, 5 strains carried KPC-2 gene.The blaNDM-1 was transferable by plasmids.Southern blot hybridization indicated that the blaNDM-1 gene was located on plasmid in size of 46 kb.The plasmid belonged to incompatibility group IncX 3.Seven types of CR-KP were detected by PFGE.In addition, MLST assigned them to sequence type ( ST)11, ST395, ST17, ST37, ST263, ST14 and ST76 types.SDS-PAGE and ompK35/36 genes sequence analysis of Omp indicated that there was absence of outer membrane proteins OmpK 36 in ST11, ST395, ST37 strains.However, the other STs strains expressed lower quantities of OmpK 36.Conclusions High level carbapenem resistance in K.pneumoniae causing infection in burn patients is attributable to production of plasmid-mediated metallo· β-laetamase NDM-1 combined with porin OmpK36 deficiency or low expression .The K.pneumoniae with NDM-1 and KPC-2 carbapenemase were detected .

BACKGROUNDProstate specific membrane antigen (PSMA) can facilitate the growth, migration, and invasion of the LNCaP prostate cancer cell lines, but the underlying molecular mechanisms have not yet been clearly defined. Here, we investigated whether PSMA serves as a novel regulator of the phosphatidylinositol 3-kinase (PI3K)/Akt signaling by employing PSMA knockdown model and PI3K pharmacological inhibitor (LY294002) in LNCaP prostate cancer cells.

METHODSPSMA knockdown had been stably established by transfecting with lentivirus-mediated siRNA in our previous study. Then, LNCaP cells were divided into interference, non-interference, and blank groups. We first testified the efficacy of PSMA knockdown in our LNCaP cell line. Then, we compared the expression of PSMA and total/activated Akt by Western blotting in the above three groups with or without LY294002 treatment. Furthermore, immunocytochemistry was performed to confirm the changes of activated Akt (p-Akt, Ser473) in groups. Besides, cell proliferation, migration, and cell cycle were measured by CCK-8 assay, Transwell analysis, and Flow cytometry respectively.

RESULTSAfter PSMA knockdown, the level of p-Akt (Ser473) but not of total-Akt (Akt1/2) was significantly decreased when compared with the non-interference and blank groups. However, LY294002 administration significantly reduced the expression of p-Akt (Ser473) in all the three groups. The results of immunocytochemistry further confirmed that PSMA knockdown or LY294002 treatment was associated with p-Akt (Ser473) down-regulation. Decrease of cell proliferation, migration, and survival were also observed upon PSMA knockdown and LY294002 treatment.

CONCLUSIONSTaken together, our results reveal that PI3K/Akt signaling pathway inhibition may serve as a novel molecular mechanism in LNCaP prostate cancer cells of PSMA knockdown and suggest that Akt (Ser473) may play a critical role as a downstream signaling target effector of PSMA in this cellular model.

Antigens, Surface ; genetics ; metabolism ; Cell Line, Tumor ; Glutamate Carboxypeptidase II ; genetics ; metabolism ; Humans ; Male ; Phosphatidylinositol 3-Kinases ; metabolism ; Prostatic Neoplasms ; enzymology ; genetics ; therapy ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA Interference ; Signal Transduction ; genetics ; physiology