Objective: To investigate the influence factors and dynamic varitation rule for the accumulation of huperzine A (Hup A) from thallus of Huperzia serrata, which could produce Hup A. Methods: The optimal concentration of plant growth substances for the growth of thallus of H. serrata were screened using orthogonal design method. The content of target extract from thallus of H. serrata were tested using HPLC. The kinetic curve of thallus growth was described using Logistic growth model. Results: The thallus of H. serrata could produce the most Hup A (72.45 μg/L) when cultured in 1/4 MS solid medium complemented with 12 h/d white light and the relative growth rate could reach 3 174.5%. According to the growth curve of thallus of H. serrata like "S" shape, the best successive generation date of callus culture of H. serrata was 50 d. The concentration of Hup A increased obviously after 60 d when the thallus reached the stable period and the maximal value of Hup A was 7.453 μg/g at 85 d, which meant that the rapid growth of thallus was priority to the accumulation of Hup A. Conclusion: The relational model between the accumulation of Hup A from thallus and its growth is non-growth conjugation. In addition, the growth law of thallus is consistent with Logistic growth model and the maximum specific growth rate (μmax) was 0.071 d-1.

In order to investigate the biological effects of the VP22?-mE6?/mE7 built-in adjuvant fusion protein vaccine on the tumor associated with HPV-16 infection.HSV-1 VP22? and HPV-16 mE6?/mE7 genes were cloned,and the pET28a-VP22?-mE6?/mE7 recombinat prokaryotic expression vector was constructed.Vector was transformed into Rosetta(DE3)E.coli string and expressed under the induction of IPTG.The re-naturalized protein was then purified via Ni2+ affinity adsorbent column and identified by SDS-PAGE and Western blot.Purified protein was immunized BalB/C and C57BL/6 mice to evaluate the immunogenicity and anti-tumor activity.The expressed recombinant protein formed as inclusion body with a prediction MW about 34kDa and contained approximately 45% of total somatic protein.The VP22?-mE6?/mE7 immunization induced higher titer of specific IgG against HPV,higher level of lymphocyte proliferation and better effect on suppressing HPV16 positive TC-1 tumor growth than the mice immunized with mE6?/mE7 alone.The results showed that the recombined built-in adjuvant vaccine could induce specific cellular immune response in vitro and inhibit the TC-1 tumor proliferation in vivo,that would be a foundation for further studies and developments of inner adjuvant vaccines.

Objective To study the influence of fluoride on the expression of osteoprotegerin(OPG) mRNA and protein in suckling rat osteoblasts. Methods Osteoblasts obtained from calvarial of suckling Wistar rats were cultured in vitro in the media supplemented with NaF at a series of doses[O(control), 1,2 and 4 mg/L groups], and OPG mRNA expression and protein were evaluated by RT-PCR and ELISA methods, respectively. Results OPG mRNA expression in suckling rat osteoblasts cultured in vitro significantly increased after exposure to NaF for 48 h and 72 h(F=333.48,808.34,P<0.05). OPG mRNA expression in suckling rat osteoblasts cultured in vitro after exposure to NaF for 48 h at different doses(0.810±0.003, 0.819±0.031 and 0.870±0.044 for 1,2 and 4 mg/L groups, respectively) compared with that of control (0.800±0.040, all P<0.05). OPG mRNA expression further increased for 72 h exposure to NaF(0.933±0.047,1.031±0.051,1.240±0.062 for 1,2 and 4 mg/L, respectively), significantly higher than that of the control (0.805±0.020,all P<0.05) and corresponding groups at 48 h. NaF doses and time exposure exhibited a significant synergistic effect on OPG mRNA expression(F=2004.16, P<0.05). NaF also enhanced OPG protein expression in suckling rat osteoblasts cultured in vitro. Significant differences were observed only in 4 mg/L group(0.228±0.014,0.277±0.048) and control(0.205±0.012,0.229±0.010) at 48 h and 72 h (P<0.05). In addition, OPG protein expression at 72 h post-exposure was higher than that at 48 h,but there was no synergistic effect between concentration and time(F=1.21,P>0.05). Conclusions The results suggested that NaF could increase OPG mRNA and protein expression in suckling rat osteoblasts with a synergistic effect between the doses and exposure time.

Objective:To investigate the underlying mechanism of histone deacetylase (HDAC) inhibitor FK228-in- duced apoptosis of the prostate cancer cell line DU145.Methods:The inhibitory effect of FK228 on DU145 cell growth and its cytotoxicity were determined by MTT assay;cell cycle arrest was detected by flow cytometry assay;morphological change was observed by Giemsa staining;and defined kinase protein levels were determined by Western blot analysis.Re- suits:FK228 obviously inhibited DU145 cells growth,arrested cell cycle at G_0/G_1 phase,induced cells morphological changes and degraded several kinase proteins,including EGFR,Her2,Raf-1,Src,Cdk4 and IAP member Survivin.The degradation of these kinases blocked Raf-Mek-Erk and PI3K/Akt survival signal pathways,inducing apoptosis.Condu- sion:FK228 may induce DU145 cell apoptosis through depletion of multiple kinase proteins and blockade of survival sig- nal pathways of DU145 cells.

Adult ; Appendiceal Neoplasms ; immunology ; pathology ; Dendritic Cell Sarcoma, Follicular ; immunology ; pathology ; Humans ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-kit ; analysis ; Receptors, Complement 3b ; analysis

Objective To explore the action mechanism of moxibustion in treating Crohn's disease (CD) by observing the effect of herbal cake-partitioned moxibustion on the expression of NLRP3 inflammasome (NLRP3, ASC and Caspase-1) and IL-1β. Method Male Sprague Dawley (SD) rats of clean conventional grade were randomized into a normal control group (NG), a model control group (MG), a herbal cake-partitioned moxibustion control group (MoxG), and a sham moxibustion control group (SMG). The CD rat models were developed by using the mixture of 2, 4, 6-trinitrobenzene sulfonic acid (TNBS) and alcohol via enema. When the models were successfully developed, the MoxG received moxibustion intervention at bilateral Tianshu (ST25) and Qihai (CV6), and the SMG were administered by unlighted moxa cones. At the end of treatment, the length of each rats' colon and the colon macroscopic damage index (CMDI) score were recorded, the histopathological variations of rats' colons were observed by adopting HE staining and light microscope, and the expressions of NLRP3, ASC, Caspase-1 and IL-1β in rats' colons were determined by using immunohistochemical technique. Result Compared to the NG, the MG had its rats' colons present with severe damages, fissured ulcers and inflammatory cell infiltration with edema, and granulomas in submucosa of some colons, and its expressions of NLRP3, ASC, Caspase-1 and IL-1β increased significantly (P<0.05);compared to the MG, the MoxG had rats' colons present with improved structures and reduced intestinal inflammation, and its expressions of NLRP3, ASC, Caspase-1 and IL-1β dropped significantly (P<0.05); the SMG had its rats' colon inflammation present similarly to the MG, and its expressions of NLRP3, ASC, Caspase-1 and IL-1β had no significant difference (P>0.05). Conclusion Herbal Cake-partitioned moxibustion can down-regulate the expressions of NLRP3, ASC, caspase-1 and IL-1β in CD rats' colons to promote the repair of colon damage.

Objective:To observe the effect of moxibustion on the protein and mRNA expressions of corticotropin-releasing factor (CRF)and corticotropin-releasing factor receptor 1 (CRFR1) in hypothalamus of trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis rats,and to explore the central mechanisms of moxibustion in improving visceral pain and the pain-related emotions in experimental colitis rats.Methods:Thirty-six Sprague-Dawley (SD) rats were randomly divided into a normal group (NG),a model group (MG),a herb-partitioned moxibustion group (HPMG) and a sham herb-partitioned moxibustion group (SHPMG).Except the NG,rats in the remaining three groups all received TNBS enema to establish experimental colitis models.The HPMG received herb-partitioned moxibustion (HPM) at bilateral Tianshu (ST 25) and Qihai (CV 6) for intervention;for the SHPMG,the herbal cakes and moxa cones were only placed on the acupoints but not ignited;rats in the MG and NG were only fixed in the same way as those in the HPMG but did not receive any treatment.At the end of the intervention,the abdominal withdrawal reflex (AWR) score,the open field test (OFT) score and the elevated plus maze (EPM) score were observed to measure the changes in visceral pain and pain-related emotions of the rats.The enzyme-linked immunosorbent assay (ELISA) was used to examine the expressions of CRF and CRFR1 proteins in hypothalamus;the fluorescence-based quantitative polymerase chain reaction (PCR) was used to detect the expressions of CRF and CRFR1 mRNAs in hypothalamus.Results:Compared with the NG,the AWR score increased significantly and the OFT and EPM scores dropped significantly in the MG (all P＜0.05),and the expressions of hypothalamic CRF and CRFR1 proteins and mRNAs increased significantly (all P＜0.01).Compared with the MG and SHPMG,the AWR score dropped significantly and the OFT and EPM scores increased significantly in the HPMG (all P＜0.01),and the expressions of hypothalamic CRF and CRFR1 proteins and mRNAs decreased significantly (all P＜0.05).There were no significant differences between the MG and the SHPMG (all P＞0.05).Conclusion:HPM can down-regulate the abnormally increased expressions of CRF and CRFR1 proteins and mRNAs in hypothalamus of the TNBS-induced experimental colitis rats,which is plausibly one of its action mechanisms in mitigating visceral pain and the pain-related emotions in the experimental colitis rats.

OBJECTIVETo study the efficacy of endotracheal lavage in neonatal ventilator-associated pneumonia (VAP).

METHODSFifty-eight neonates with VAP between January 2002 and December 2008 were randomly assigned to two groups: lavage and control (n=29 each). After withdrawal from ventilator, both groups received sensitive antibiotics therapy according to sputum culture results as well as supportive treatment. The lavage group was additionally treated with endotracheal lavage (2-3 times daily). The therapeutic effects were compared between the two groups.

RESULTSThere were no significant differences in the average time of mechanical ventilation between the lavage and the control groups. The effective rate in the lavage group (93%) was significantly higher than that in the control group (69%; p<0.05). Three percent of patients in the lavage group required twice or more mechanical ventilation compared with 24% in the control group (p<0.05). Blood gas analysis results were obviously improved in the lavage group 2 hrs after treatment (p<0.01).

CONCLUSIONSEndotracheal lavage can decrease the number in mechanical ventilation and improve therapeutic effects in neonates with VAP.

Female ; Humans ; Incidence ; Infant, Newborn ; Male ; Pneumonia, Ventilator-Associated ; epidemiology ; therapy ; Respiration, Artificial ; statistics & numerical data ; Therapeutic Irrigation ; methods ; Trachea

AIMTo study the dissolution rate of solid pharmaceutical preparation on-line, a multiple channel fiber-optic chemical sensor based on fluorescence multiple quenching (FOCSMQ) without filtering and sampling was made.

METHODSUsing the multiple channel FOCSMQ linked with computer, the dissolution rates of ofloxacin tablets, metronidazole tablets and nitrofurantoin tablets were monitored continuously on-line. The instrument can give the sample data, display the real time curve and calculate the T1/2 and td automatically. A computer was used to select the best function from five common fitting models to fit the dissolution curve.

RESULTSThe average recoveries of the FOCSMQ method were 97.4%-104.4%, 97.4%-103.8% and 96.6%-102.1%. The RSDs (n = 6) of within-day and between-day were less than 5%. The parameters of the dissolution and all results of measurement using the instrument have no significant difference compared with the Chinese Pharmacopoeia (ChP) (2000) method and the United States Pharmacopoeia (USP) (23) method (P > 0.05). It does not need sampling and dilution, and never contaminate sample. It can shorten time of the experiment.

CONCLUSIONThe method is simple, rapid and reliable.

Chemistry, Pharmaceutical ; instrumentation ; Fiber Optic Technology ; methods ; Metronidazole ; chemistry ; Nitrofurantoin ; chemistry ; Ofloxacin ; chemistry ; Optical Fibers ; Solubility ; Tablets ; chemistry ; Transducers

Objective To learn the main pathogenic microorganisms,and to acquire the data of pathogens composition anddrug resistance of infant diarrhea disease in Wenzhou City.Methods The diarrheal stool specimens of <5 years oldchildren were collected from outpatients and inpatients in children′s hospital and were cultured and detected.Results Atotal of 201 strains pathogenic bacteria were separated from 51 7 clinical specimens,and the rate of total detection is38.88%,of which included 74 strains intestinal pathogenic bacteria and 1 27 strains diarrhea virus,with the rate of detection1 4.31 % and 24.56%,respectively.Among intestinal pathogenic bacteria,the E.coli was the most common bacteria(36strains,48.65%),followed by salmonella(31 strains,41 .89%).Diarrhea virus included norovirus type 2 (56 strains,44.09%),norovirus type 1 (24 strains,1 8.90%),A group rotavirus (56 strains,31 .50%)and goblet virus(7 strains,5.51 %).The resistance rate of Salmonella to ampicillin was 64.52%,and that to ampicillin,tetracycline,compound newMing were more than 70.00%.The resistance rate of Rifampicin.Novobiocin in all bacteria reached 1 00%.Conclusion E.coli and salmonella epidemic strains were the common pathogenic bacteria in infectious diarrhea among infants inWenzhou City.The predominant viruses were norovirus and rotavirus group A.The drug resistance of various pathogenicbacteria was different,and active surveillance should be strengthened.