Objective:To analyze the genetic sequence characteristics of amomum viosum and establish a rapid identification meth-od for amomum viosum by fluorescent quantitative PCR based on DNA analysis. Methods:Amomum viosum and the other samples be-longing to the same genera were collected and identified by experts in the domain. DNA was isolated using commercial kits. The prim-ers and probe were designed according to the conserved region of ITS in amomum viosum. The reaction conditions were optimized to es-tablish the fluorescent quantitative PCR method for the rapid detection of amomum viosum. Results:The fluorescent quantitative PCR method for the rapid detection of amomum viosum was set up. The method could identify amomum viosum successfully, while those samples in the same genera were without amplification curves. Conclusion: Amomum viosum can be identified rapidly by fluorescent quantitative PCR besides the traditional identification by experts.

ObjectiveTo assess the degree of pain in elderly patients with mechanical ventilation in ICU using critical-care pain observation tool(CPOT) and to choose the correct sedative and analgesic method.Methods 110 elderly patients in ICU after neurosurgery were divided into three assessment stages,every stage had two record points and total six points (T1-T6):the first stage (intubation and unconsciousness,T1-T2),the second stage (intubation and consciousness,T3T4 ) and the third stage(extubation and consciousness,T5-T6 ).Among them T1,T3and T5were nonnocuity assessment points of every stage,while T2,T4 and T6 were nocuity assessment points of every stage.The assessment time was one minute at every point.After recorded at every point in second and third stages,patients were asked to use the pain intensity descriptive scale (PIDS) themselves.CPOT,heart rate and mean arterial pressure (MAP) from T1 to T6 were recorded as well as PIDS from T3 to T6 in second and third stages.Results In the three stages,CPOT〔(26.8 vs.0.54,3.36 vs.1.20,2.78 vs.0.68) scores〕,HR〔(95 vs.85,94 vs.82,94 vs.84)beat/min〕 and MAP〔(95 vs.85,95 vs.87,94 vs.87)mm Hg〕 at T2,T4and T6 were higher than T1 (t=-42.89,-55.95,-55.38),T3 (t =- 5.52,- 11.33,- 11.78) and T5 ( t =- 5.54,- 9.95,- 11.33 ) ( P＜ 0.05 ).The PIDS at T4 and T6were higher than at T3and T5in the second and third stages 〔(2.52 vs.1.69,2.12 vs.1.44)scores〕 (P＜0.05).The correlation coefficient between CPOT and PIDS at T3 and T4 in the second stage were 0.49 and 0.58,respectively (P＜0.05),and between CPOT and PIDS at T5 and T6 were 0.52 and 0.59 in the third stage,respectively (P ＜ 0.05),and they both reached moderate correlation.ConclusionsCPOT may be an effective way to assess the degree of pain in elderly patients with mechanical ventilation at present.

Anti-Bacterial Agents ; adverse effects ; Child, Preschool ; Clindamycin ; adverse effects ; Humans ; Infant ; Male ; Paraplegia ; chemically induced

Objective To observe the effect of propofol on phosphorylation of a-amino-3-hydroxy-5-methyl-4-isoxa-zolep-propionate receptors (AMPARs) GluR1 subunit and long-term potentiation (LTP) in cultured hippocampal neurons in aged rats.Methods A total of 30 18-month-old rats were decapitated,the brains were rapidly removed and hippocampal slice were prepared.The slices were randomly divided into control group (perfused with artificial cerebrospinal fluid,n=10),propofol-treated group (perfused with propofol in artificial cerebrospinal fluid,n=10)and propofol+ phorbol-12-myristate-13-acetate (PMA)-treated group (perfused with propofol and phorbol ester in artificial cerebrospinal fluid,n=10).Extracellular excitatory postsynaptic potentials (EPSP) were recorded from the CA1 region of hippocampal slices.After perfusion for 20 min,LTP was induced using higher-frequency stimulation (HFS,100Hz,400 pulse) by the Schaffer-collateral pathway.The phosphorylation of AMPA-GluR1 subunit was assayed in cultured rat neurons by Western blot.Results The value of EPSP in propofol-treated group (105.50 ± 3.77) was much lower than in control group (242.10±14.68) and in propofol+ PMA-treated group (239.40±8.98) (F=2.90,P＜0.05),and there was no significant difference in the value of EPSP between control group and propofol+ PMA-treated group (P＞0.05).The level of P-Glu1/Glu1in propofol-treated group (0.68±0.15) was much lower than in control group (1.67±0.20) and in propofol+PMA-treated group (1.57±0.18) (F=6.84,P＜0.05),while there was no difference in the level of P-Glu1/Glu1 between control group and propofol + PMA-treated group (P ＞ 0.05).There was no difference in the value of GluR1/β-actin among the three groups (F=0.31,P＞0.05).Conclusions Propofol possesses the ability to inhibit LTP induction and attenuate AMPA receptor GluR1 subunit phosphorylation through modulation of PKC pathway.

Objective To construct a eukaryotic expressing vector harboring human DC-SIGN, and establish a BHK21 cell line stably and highly expressing DC-SIGN. Methods The DC-SIGN gene fragment which contained Not I and BamH I sites was amplified by PCR from pUNO-hDCSIGN1Aa plasmid, digested with Not I and BamH I, and then cloned into an eukaryotic expression vector pIRES-neo to construct eukaryotic expression vector pIRES-neo-DC-SIGN. The recombined plasmid was identified with Not I and BamH I enzyme digestion and sequencing, the latter was then transfected to BHK21 cells by LipofectamineTM 2000. After screening culture by G418, BHK21 cell line stably expressing DC-SIGN was established. The expression of DC-SIGN was identified by flow cytometry, Western blotting and immunofluorescence method. Results The gene sequence of DC-SIGN was consistent with that of design. PCR and double enzyme digestion analysis showed that the recombinant plasmid pIRES-neo-DC-SIGN was constructed successfully. After transfection, positive clones were selected with G418. After limiting dilution assay, BKH21 cell lines stably expressing DC-SIGN were established. The detection result of flow cytometry showed that the expression ratio of DC-SIGN positive clones was close to 90%. The result of immunofluorescence displayed that the expression of DC-SIGN was mostly located on the surface of cell membrane. Western blotting displayed the specific band of DC-SIGN protein. It showed that the BHK21 cells stably expressing DC-SIGN were successfully established. Conclusion DC-SIGN eukaryotic expression vector has been successfully constructed. The successful establishment of BHK21 cell lines which can stably express DC-SIGN provides a substantial foundation for further study on the DC targeting vaccines.

Objective To study the diagnosis, treatment and prognosis of mixed thyroid malignant tumors. Methods Clinical data of 7 cases with merged different histological types of thyroid malignant tumor treated from January 1977 to December 2006 were retrospectively analyzed. Results Merged different histologic types of thyroid malignant tumor accounted for 0. 14% of all thyroid malignant tumors treated during this period. Preoperative imaging and laboratory data had no specific value in the diagnosis of this merged different histologic types of thyroid malignant tumors. Radical resection in combination of hormonal therapy and 131I radiotherapy achieved a satisfactory result, though thyroid malignant tumor combined with thyroid cancer usually predict a poor prognosis. Conclusions Merged different histologic types of thyroid malignant tumor is a rare clinical entity, with the pathogenesis being obscure and no consensus of opinion on its nomenclature. The prognosis depends on the highest ～ade among an individual group of malignant tumors.

AimTo evaluate the efficiency of gene amplification technique used in detecting the specimens colleted from rodents to identify natural epidemic foci of scrub typlus. MethodMice of Kunining strain were experimentally infected by a certain amount of Oriential tsutsugamushi. The specimens of blood clot and spleen from the infected animals were detected by nested polymerase chain reaction(nPCR)specific to O. T sutsugamush at the day 3,6 and 9 of post-infection. Then the technique was used for detection of samples collected from field. As an infected index ,the specimen was considered to be positive only if a 88-bp DNA fragment from Sta 58kDa gene of O. Tsutsugamushi could be produced. According to the study ,it was estimated whether or not that the sampling area is a natural epidemic focus of the disease. ResultsThe specimens of both blood clot and spleen from the mica at day 3 of post-infecction showed negative to the specific PCR product ,but positive when detected at day 6 and hereafter. Of 111 spleen samples from the field collections in the northwest of Fujian province,one was positive, and another positive sample was in the 29 blood clots from Jiangxi province. It is demonstrated that these areas have been the natural epidemic foci. Conclusion The nPCR method is of highly sensitive and specific to be used in the etiologic study on specimens from field rats.

Recently, microRNAs (miRNAs) have been shown to be involved in multiple biological pathways that can influence tumor progression and metastasis and they can serve as prognostic biomarkers in many cancers. The present study examined the prognostic significance of miR-215 in cervical cancer. The paraffin-embedded paired cervical scrape samples and tumor tissue samples from 302 patients with stage II cervical cancer were detected for the expression of miR-215 by using qRT-PCR. A miR-215-based classifier was established by using the Cox regression model. The prognostic and predictive accuracy of this classifier was determined in both the internal testing group of 138 patients, and the external independent group of 280 patients. Moreover, cervical cancer HeLa cells overexpressing miR-215 (HeLa-miR-215) were constructed and subcutaneously injected into the nude mice to examine the effect of miR-215 on tumor growth and metastasis in vivo. The results showed that the expression level of miR-215 was significantly higher in cervical cancer tissues than in paired normal tissues (P<0.0001). When patients were classified into high- and low-risk cancer progression groups according to miR-215 level, the 5-year disease-free survival in high- and low-risk groups were 43% (95% CI: 32.1-51.6) and 67% (95% CI: 48.6-77.3) (hazard ratio [HR] 2.02, 95% CI: 1.16-3.52; P=0.013) respectively. Moreover, the expression level of miR-215 was negatively associated with survival rate in patients at TNM stage T3 (HR: 3.317; 95% CI: 1.18-5.14, P=0.017) and TNM stage T4 (HR: 3.48; 95% CI: 1.49-4.45, P=0.008). Tumor volume in nude mice injected with HeLa-miR-215 cells was significantly larger than that in mice injected with control HeLa cells. It was concluded that the expression level of miR-215 is associated with cervical tumor progression and worse survival rate, suggesting that it may serve as a potential prognostic marker to identify patients at higher risk of recurrence.

After percutaneous coronary intervention (PCI) was performed,since CHD risk factors still exist, coronary restenosis rate remains high.Therefore, self-management after PCI is very important.The present article made a review on current condition and research progress of self-management in patients after PCI, aiming at providing reliable evidence for rehabilitation after PCI.

With the coming of knowledge economics, technical innovation is becoming more and more important.However, the risk accompanied to the technical innovation exists everywhere and is a serious problem demanding prompt solution.Facing the realistic condition, a new idea on the application of vague comprehensive evaluation model to investigating the technical risk in Chinese materia medica innovation project has been put forward.According to the properties of new drug project and the vague mathematic theory, the determined nature issue has been quantified, what happened described objectively, and a model of vague comprehensive evaluation constructed increasingly.The result provides a prerequisite for investment and decision of the technical project.By the analysis of risk factors it is easy and aggressive to control and avoid the risk during the process of project implement.Generally the vague comprehensive evaluation model can be available and benefit to adjust the investment structure and to enhance the scientific decision level.