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.

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.

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

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 investigate the relationship between pure-tone average (PTA), the fractional anisotropy (FA) of the auditory pathway, cognitive cortex and auditory cortex in presbycusis. METHOD: Twenty-five elderly subjects with presbycusis were participated in the study. PTA, speech discrimination abilities were evaluated in each subject. Diffusion tensor imaging (DTI) was applied to access the FA of the IC, the superior frontal gyrus and the Heschl's gyrus. Compare the difference between two sides of the values of FA in the three areas. Bivariate correlation analysis was performed to evaluate the effects of PTA and FA of the inferior colliculus (IC), the superior frontal gyrus and the Heschl's gyrus on speech discrimination abilities. RESULT: There were no significant differences between the left and right side of the inferior colliculus (P > 0.05). Higher FA values were recorded at the left side of the Heschl's gyrus and the superior frontal gyrus (P < 0.05). Both PTA and the FA of the superior frontal gyrus have a negative association with speech discrimination abilities (P < 0.01, P < 0.05), while the FA of the Heschl's gyrus has a positive association with speech discrimination abilities (P < 0.05). CONCLUSION Our findings indicated that the speech discrimination abilities of the elderly is not only related to the peripheral auditory function, but also to the central auditory and cognitive function.
Aged ; Audiometry, Pure-Tone ; Auditory Cortex ; physiopathology ; Auditory Pathways ; Diffusion Tensor Imaging ; Humans ; Inferior Colliculi ; physiopathology ; Presbycusis ; diagnosis ; Speech Perception

Objective To investigate the effects of stellate ganglion block (SGB) on erythrocyte immunity in patients with acute cerebral infarction.Methods Twenty-four patients (13 male, 11 female) who developed acute cerebral infarction for less than 3 days were randomly divided into 2 groups (n=12each): Group A receiving traditional treatment and Group B receiving traditional treatment + SGB.The patients ranged in age from 51 to 64 yr and weighed 52-71 kg. All patients received intravenous 5% glucose 25 ml plus citicoline sodium 1.0 g and sodium ozagrel injectio 250 ml daily for 10 days in addition to dehydration and effective control of complications and intracranial pressure. Group B received SGB on one side alternatively with 1% licocaine 10 mi once a day for 10 days. Fasting venous blood samples were taken in the early mornings of the day before treatment (baseline, T1 ) and the 1st, 5th and 10th day of treatment (T2-4) for determination of the plasma MDA concentration and SOD activity, erythrocyte C3b receptor rosette rate (RBC-C3bRR) and RBC immune complex rosette rate (RBC-ICR) and Ne+-K+-ATPase activity in erythrocyte membrane.Results The plasma MDA concentration and RBC-ICR were significantly decreased during treatment es compared with the baselines at T1 in both groups (P＜0.05 or 0.01), but were significantly lower in Group B than in Group A (P＜0.05 or 0.01 ).The activities of plasma SOD and Na+ -K+ -ATPase in erythrocyte membrane and RBC-C3bRR were significantly increased during treatment as compared with the baselines at T1 and were significantly higher in Group B than in Group A.Conclusion SGB combined with traditional treatment can increase the activities of plasma SOD and Na+ -K+ -ATPase in erythrocyte membrane, inhibit production of oxygen free radicals and enhance RBC immune function in patients with acute cerebral infarction.

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.

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.