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.

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.

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 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 explore the value of high frequency ultrasonography for the early diagnosis of hyperuricemia in patients with joint disease.Methods Ninety-eight patients with hyperuricemia and 100 healthy persons,according to with or without history of acute gout attack,were divided into symptomatic group,asymptomatic group and control group,whose first metatarsophalangeal joint,ankle and knee were examined by high frequency ultrasonography.Chi-square test was used for statistical analysis.Results ① The joint lesions detection rate in the symptomatic group,asymptomatic group and the control group was 57％ (13/23),16％ (12/75),0 (0/100) respectively.There was statistical significant difference between the symptomatic group and the asymptomatic group (x2=9.69,P＜0.05).② The symptomatic group had 29 joint involvement (29/138),including 25 at the first metatarsophalangeal joint,3 at the ankle and one at the knee joint.The asymptomatic group had 14 joints involved (14/450),which were all located at the first metatarsophalangeal joint.③ The sonographic appearance of the two groups of arthropathy were synovial thickening,effusion,crystal deposition andbone erosion.The symptoms group presented as tophi,and increased synovial blood flow.Conclusion Patients with asymptomatic hyperuricemia may have joint diseases.The joint disease detection rate of the symptomatic group is higher than that of the asymptomatic group.High frequency ultrasonography can be used a conventional imaging method for the screening of patients with hyperuricemia joint disease.The focus of the screening of asymptomatic patients is the first metatarsophalangeal joint.The focus of the screening of symptomatic patients is the first metatarsophalangeal joint and the joints that were attacked in the past history.

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.

Objective To investigate the clinical effect of early rehabilitation treatment on patients with severe traumatic brain injury (sTBI).Methods Forty sTBI patients were divided into treatment group (n =20) and control group (n =20) according to the random number table.Conventional treatment was performed on all patients including dehydration to decrease intracranial pressure,hemorrhage control,neurotrophic treatment,antiinflammation therapy,and gastric acid control.In addition to these interventions,patients in treatment group received hyperbaric oxygen treatment,median nerve stimulation,fastigial nucleus stimulation,and bedside motor therapy in the early period.Intracranial pressure and partial pressure of brain tissue oxygen (PbtO2) were continuously monitored during the process of treatment.GCS was measured before and 15 days after treatment and single-photon emission computed tomography (SPE-CT) was used to evaluate cerebral perfusion.Results There was no statistical difference between the two groups with respect to GCS in advance of treatment (P ＞ 0.05),but GCS differed between treatment group and control group after treatment [(10.18 ± 3.75) points vs (8.33 ±2.36) points,P ＜0.05],with substantial improvement in treatment group.Significantly improved cerebral perfusion was seen in treatment group.On day 5 after treatment,intracranial pressure in treatment group lowered significantly compared with that in control group (P ＜ 0.05).On day 6 after treatment,PbtO2 was significantly higher in treatment group than in control group (P ＜ 0.05).Conclusion Early rehabilitation treatment leads to improved outcome and acts a positive effect on nerve function recovery.

Objective To study the effect of astragaloside on proliferation and apoptosis in human keloid fibroblasts.Methods The human keloid fibroblast ceils were treated with different concentration of astragaloside(10、20、40 ng/mL).Cell proliferation was detected by MTT,the gene expreesion levels and protein levels of apoptosis-related proteins,survivin,p53 and Bcl-2.were determined by real-time PCR and Western blot,respectively.Results Comparecl with control group(treated with 0 ng/mL astragaloside),the absorbance values (A490 nm) of each concentration group were significantly reduced,which suggest that the proliferation of all keloid fibroblast were markably inhibited in a dose-dependent way (P＜0.05).The gene expreesion levels and protein levels of apoptosis-related proteins,survivin、Bcl-2 were largely suppressed and P53 werelargely promoted in a dose-dependent.Conclusion The keloid fibroblasts cells proliferation and apoptosis could be regulated by astragaloside.

Objective To further investigate the association among clinical pathology,complement activation and renal secretory IgA (SIgA) deposition in patients with IgA nephropathy (IgAN).Methods The activation of serum complements were detected by immunoturbidimetry and ELISA.Renal deposition of SIgA and activation of complements were detected by immunofluorescence.Then the association among clinical pathology,complement activation and renal SIgA deposition were analyzed in IgAN patients.Results In all 201 patients with IgAN,59 patients had SIgA deposition with higher incidences of mucosal infection history and hematuria (P ＜ 0.05),lower levels of serum cystatin C,β2 microglobulin and lower tubulointerstitial lesion grades and T-grade in the Oxford classification (P ＜ 0.05),when compared with patients without SIgA deposition.Both alternative and mannose binding lectin (MBL) pathways were activated in patients with or without SIgA deposition.Patients with MBL pathway activation had lower estimate glomerular filtration rate (P ＜ 0.01),higher serum creatinine,higher proportion of glomerulosclerosis and S-grade in the Oxford classification,more severe tubulointerstitial lesion (P ＜ 0.05).Conclusions Compared with patients without SIgA deposition,patients with SIgA deposition have a stronger link to mucosal immune.The deposition of SIgA is associated with different clinical and pathological manifestations;however,the complement activation is similar in both groups of patients.Patients with MBL pathway activation show more severe kidney injury.