Acute Disease ; Adult ; Ethylene Dichlorides ; poisoning ; Female ; Humans ; Male ; Neurotoxicity Syndromes ; therapy

To establish a convenient and rapid method for identification of Pseudostellariae Radix by molecular identification, the rDNA-ITS sequences of Pseudostella riaheterophylla and its adulterants had been aligned to find out specific fragment. The specific primers against the fragment were designed and the PCR amplification conditions were optimized. The fluorescence reaction of the PCR products colored by 100 x SYBR Green I was observed under UV. The concentration of reaction buffer included 5.5 μL DNA Taq polymerase premix, 10 pmol Tzs-2F and 10 pmol Tzs-2R, 20-80 ng template DNA, and plus double sterile distilled water to 25 μL. The PCR thermal profile was as follows: predenaturation at 95 degrees C for 1 min, followed by 30 cycles of denaturation at 95 degrees C for 5 seconds, primer annealing and extension at 56 degrees C for 15 seconds, then it was extension at 72 degrees C for 30 seconds. The fluorescence reaction of Pseudostellariae Radix showed green fluorescence, while adulterants had not. Extraction, amplification DNA and all steps of molecular identification could be completed successfully in 40 minutes. The approach could amplify DNA template of Pseudostellariae Radix specificity, and its product with 1 μL 100 x SYBR Green I could engender green fluorescence under UV. The method was simple and accurate, so it could be used for identification of Chinese traditional medicine.
Caryophyllaceae ; classification ; genetics ; DNA Primers ; genetics ; Drug Contamination ; prevention & control ; Plant Roots ; classification ; genetics ; Polymerase Chain Reaction ; methods

Objective: To observe the dynamic changes of plasma level thymosinβ4 (Tβ4) in acute myocardial infarction (AMI) patients with intervening therapy within 15 days of onset and to explore the relationship between Tβ4 and clinical prognosis in AMI patients.
Methods: Our research included 2 groups:AMI group, n=69 and Control group, the patients with suspected chest pain while CAG excluded coronary artery stenosis, n=32. Plasma levels of Tβ4 were examined in all AMI patients on admission day and every day until 15 days of onset;AMI patients were followed-up for 18 months and the endpoint was defined as major adverse cardiovascular event (MACE) occurrence.
Results: ①Compared with Control group, AMI group had increased plasma level of Tβ4 on admission day and on day-15 of onset, P<0.01. ② With intervening therapy, AMI group had elevated Tβ4 level upon immediate onset, it was decreased on day-1, reached low level on day-3 and elevated to peak on day-6, then reduced followed by slightly raising on day-11.③During follow-up period, the AMI patients without MACE had the higher mean in-hospital maximum Tβ4 value than those with MACE occurrence, P<0.01. Logistic regression analysis indicated that the mean in-hospital maximum Tβ4 value was related to MACE occurrence during follow-up period (OR=0.999, 95%CI 0.999-1.000).
Conclusion: AMI may induce up-regulated expression of plasma Tβ4;with intervening therapy, Tβ4 showed a trend of“elevation-reduction-elevation-reduction”at the early stage of AMI. High expression of Tβ4 was helpful for improving clinical prognosis in AMI patients which may provide a theoretical basis for exogenous use of Tβ4 in AMI treatment.

AIMTo study the effect of proanthocyanidins on the COX-2 enzyme activity and COX-2 protein expression in LPS-induced RAW264.7 cells.

METHODSAfter being pretreated with different concentrations of proanthocyanidins for 30 min, and then 1 mg x L(-1) LPS for 9 h, the effect of proanthocyanidins on the activity of COX-2 enzyme in RAW264.7 cells was analysed by radioimmunoassay (RIA). After being pretreated with different concentrations of proanthocyanidins for 30 min, and then 1 mg x L(-1) LPS for 9 h, the effects of proanthocyanidins on the expressions of COX-2 mRNA and protein in RAW264.7 cells were analysed by RT-PCR and Western blotting.

RESULTSThe activity of COX-2 enzyme was not inhibited by proanthocyanidins (0. 8, 4 and 20 mg x L(-1), P > 0.05 vs LPS group), but the activity of COX-2 enzyme was significantly inhibited by 10 micromol x L(-01) NS-398 (P < 0.01 vs LPS group). The expression of COX-2 mRNA was inhibited by proanthocyanidins (0. 8, 4 and 20 mg x L(-1)). The expression of COX-2 protein was inhibited by proanthocyanidins (4 and 20 mg x L(-1)).

CONCLUSIONProanthocyanidins had no effect on the activity of COX-2 enzyme in LPS-induced RAW264. 7 cells. Proanthocyanidins inhibited significantly the expression of COX-2 mRNA and protein in LPS-induced RAW264.7 cells.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Cell Line ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Lipopolysaccharides ; Macrophages, Peritoneal ; cytology ; enzymology ; Mice ; Proanthocyanidins ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics

The nucleocapsid protein (N) is a major structural protein of coronaviruses. The N protein of bat SARS-like coronavirus (SL-CoV) has a high similarity with that of SARS-CoV. In this study, the SL-CoV N protein was expressed in Escherichia coli, purified and used as antigen. An Indirect Enzyme-Linked Immunosorbent Assay (indirect ELISA) was developed for detection of SARS- or SL-CoV infections in bat populations. The detection of 573 bat sera with this indirect ELISA demonstrated that SL-CoVs consistently circulate in Rhinilophus species, further supporting the proposal that bats are natural reservoirs of SL-CoVs. This method uses 1-2 μl of serum sample and can be used for preliminary screening of infections by SARS- or SL-CoV with a small amount of serum sample.

Objective To investigate the radiosensitization effects of the combination treatment of clioquinol (CQ) and zinc on human cervical cell line HeLa in vitro.Methods Cells were divided into the 4 groups:controls,drug,radiation,and combined drug and radiation group.Cytotoxic effect of CQ and zinc on cell viability was determined by CCK-8 assay.Radiosensitization effect of CQ and zinc on HeLa cells was detected by colongenic assay,and the single-hit multi-target model was used to stimulate the doseresponse curve of survival and to calculate radiosensitization parameters.The cell cycle and apoptosis of HeLa cells were analyzed with flow cytometry.Luciferase reporter assay was used to study NF-κB activity of HeLa cells.Results The combination of CQ and zinc inhibited cell growth in a dose-dependent manner (F =188.00,P ＜ 0.01).The mean lethal dose was 3.16 and 2.04 Gy for radiation group and combined drug and radiation group,respectively,and hence the SER was 1.55.Compared with the radiation group,the ratio of G2-phase cells in the combined drug and radiation group decreased(t =10.39,P ＜ 0.05),the apoptosis rate increased at 24 h post-irradiation (t =5.64,P ＜ 0.01),and the NF-κB activity decreased (t =21.42,P ＜ 0.05).Compared to the control group,the NF-κB activity increased in the radiation group(t=6.23,P＜0.05),but decreased in the drug group(t =12.48,P＜0.05).Conclusions The combination of CQ and zinc could increase the radiosensitivity of HeLa cells by decreasing the ratio of G2-phase cells,increasing apoptosis and the inhibiting of NF-κB activity.

Objective To investigate the clinical states of enuresis children companied with spina bifida occulta(SBO)and study the efficient way of treatment.Methods The children with SBO were check out by X ray from a total of 121 children with bedwetting.Their parents were asked to complete the enuresis questionnaires.Urine routine test and B-ultrasound examination about kidney,bladder and ureter were also asked to be done.The clinic data of the 49 children were attained and analyzed.They were randomly divided into 2 groups,and given the controlled treatment.Group A[used 1-deamino-8-D-arginine vas-opressin(DDAVP)only] and group B(used DDAVP plus oxybutynin plus bladder training)treated for 12 weeks.Results There were totally 49 bedwetting children companied with SBO,and most of them(44 cases,89.8%)were severe type(bedwetting times≥7 times/week).Some of them coexisted with frequency,urgency,gentle urgency incontinence and microscopic hematuria(22 cases).Thirty cases were found the functional bladder capacity(FBC)decrease by B-ultrasound.The cure rates were 58.3%(group A)and 88.0%(group B)respectively.The relapse rates were 36.8%(group A)and 12.5%(group B)respectively after stopping treatment for 3 months.Conclusions SBO accounts for considerably higher rate in enuretic children.It might cause the disability of bladder function.The treatment plan with DDAVP plus oxybutynin plus bladder training can not only increase the cure rate but also lower the relapse rate.

e chain-link flap is the good way in the treatment of large areas of the lower leg soft tissue defects.

Objective To explore clinical characteristics of diagnosis and treatment of solid-cystic papillary tumor(SCPT) of the pancreas in children.Methods There were retrospectively analyzed about the 7 patients treated in our hospital for SCPT,with the ave-rage age of 11.5 years.All patients complained abdominal pain following a trauma(71.4%) or overeating(28.6%).The main presentation was abdominal mass.Of the 7 patients,4 received distal pancreatectomy,2 pancreatico-duodenectomy,and 1 only biopsy.Results The nicks of all patients were primany hed.All patients were pathologically confirmed as SCPT after operation.All patients were followed up for 4 monthes to 4.5 years,the recent result was well.Conclusions SCPT is a low-grade malignant tumor,which is often asymptomatic,but the patients with symptoms generally suffer from an abdominal mass or abdominal pain.The prognosis is excellent after operation.

OBJECTIVETo analyze the expressed mRNA of the factor subunit A (FA) in monocyte in a hereditary factor (F) deficiency family.

METHODSThe F A mRNA of the proband and the other family members was analyzed by RT-PCR, semi-quantitative RT-PCR, cloning and sequencing. The three dimensional structure of the protein was predicted by SWISS-MODEL and viewed by RASMIOL.

RESULTS(1) A large in frame deletion from codons 11 to 279, spanning from exon 2 to 7 of F A (DelCD11-279), was identified in the proband at mRNA level and a truncated protein is predicted composed of 464 amino acids. Compared with the normal and the other families, the proband showed lower level of F A mRNA in RT-PCR. (2) SWISS-MODEL analysis showed that the truncated protein lacked the β-sandwich and a part of catalytic core, resulting in loss of the normal catalytic domains.

CONCLUSIONDelCD11-279 of F A mRNA is associated with hereditary F deficiency. The reduced expressing level of F A gene is one of the causes resulting in F deficiency in the patients.

Adolescent ; DNA Mutational Analysis ; Exons ; Factor XIII ; genetics ; Factor XIII Deficiency ; genetics ; Female ; Humans ; Male ; Pedigree ; RNA, Messenger ; genetics ; Sequence Deletion