ABSTRACT
Objective@#To investigate the dynamic changes of antibodies induced by leptospiral vaccines.@*Methods@#Antigens for antibody detection were screened out. ELISA was used to analyze antibody responses induced at different time points after immunizing guinea pigs with different batches of leptospiral vaccines from different manufacturers. To investigate the relationship between antibody responses induced by leptospiral vaccines and their protective effects in animal model, guinea pigs were challenged with Leptospira after immunization.@*Results@#There was no significant antigen-antibody reaction between the LigA protein or Patoc Ⅰ antigen and the serum samples of guinea pigs immunized with leptospiral vaccines. Notable IgG and IgM antibody reactions were observed in all vaccination groups when using bacterial proteins from seven Leptospira reference strains which were used for the preparation of leptospiral vaccines as envelope antigens. Antigen-specific IgG antibodies peaked at 35 d after the last immunization, and the highest peak of antigen-specific IgM antibodies was reached 11 d after the last immunization. Results of the challenge test showed that non-diluted leptospiral vaccines induced significant IgG and IgM antibody reactions in guinea pigs as compared with those diluted three or nine times, showing good protective effects.@*Conclusions@#Analysis of the dynamic changes of antibodies induced by leptospiral vaccines revealed that there was correlation between the induced serum antibody responses and the protective effects. This study provided reference for further study on alternative methods for evaluating leptospiral vaccine potency.
ABSTRACT
Objective To analyze the correlation among 3 kinds of detection methods of routine microscopic examination,fluorescence PCR nucleic acid amplification and fungal color culture in the fungal detection of vaginal secretion.Methods The patients with suspected vaginitis treated in this hospital during 2014-2016 were selected.Each 500 cases of negative and positive vaginal secretion samples by microscopic examination were collected.The candida types were identified by using the fluorescence PCR nucleic acid amplification,then 100 samples with the positive results of fluorescence PCR nucleic acid amplification for detecting fungal were performed the fungal microbial culture to verify the accuracy rate of typing results.Results The Kappa value of consistency test between fluorescence PCR nucleic acid amplification and routine microscopic examination was 0.632,the consistency between them was poor.Among 500 positive samples of vaginal secretion detected by routine microscopic examination,382 cases (76.4 ％) of Candida albicans infection were detected by fluorescence PCR nucleic acid amplification,73 cases (14.6％) were Candida glabrata infection,10 cases (2.0 ％) were Candida tropicalis infection,3 cases (0.6 ％) were Candida albicans combined Candida glabrata infection and 32 cases (6.4 ％) were other fungal infection.Among 500 negative samples by conventional microscopic examination,152 positive cases were identified by fluorescence PCR nucleic acid amplification,including 130 cases of Candida albicans,16 cases of Candida glabrata and 6 cases of Candida tropicalis.There was no statistical significant difference in positive rate between the fluorescence PCR nucleic acid amplification and CHROMAgar rapid color method (x2 =0.131,P =0.936).Conclusion For the patients with clinical manifestations and negative microscopic examination results,it is recommended to conduct fluorescence PCR nucleic acid amplification rapid type identification or fungal culture identification.
ABSTRACT
Objective To identify mutations of CACNA1A gene in a family with hemiplegic migraine.Methods Total genomic DNA was extracted from a family with 3 affected members and 1 000 healthy controls.The proband and his patient sister were subjected to exome sequencing.Ten family members including 3 patients were subjected to linkage analysis.The coding exons of the CACNA1A gene were amplified and sequenced in affected and normal individuals. Bioinformatics analysis were performed.Results A novel CACNA1A mutation was identified in the 3 patients.The nonsense mutation of A to G was detected at nucleotide 1168 ( c.1168A >G) which converted the Asn codon ( AAT) to Asp (GAT) in exon 8.Conclusion The mutation(N390D) detected in the present study is considered to result in the Chinese Hemiplegic migraine family.
ABSTRACT
Objective To investigate the relationship between familial hemiplegic migraine (FHM ) with the mutation of CACNA1A gene .Methods Total genomic DNA of a family affected members and 1 000 normal controls was extracted for conduc‐ting the CACNA1A gene sequencing research and the bioinformatics analysis .Results The novel mutation site c .1168A>G of ex‐on located in CACNA1A gene led to Asn to be replaced with Asp (N390D) .Conclusion The mutation(N390D) of CACNA1A gene is a newly found novel pathogenic mutation lead to familial hemiplegic migraine .
ABSTRACT
Objective To study the effects of Xiaohuan Decoction on the locomotor activity, spatial learning and memory, and the serotonin (5-HT) in striatum in schizophrenia model rat, and the possible mechanism of Xiaohuan Decoction on schizophrenia.Methods The rats were randomly assigned into four groups: normal group,model group, Xianhuan Decoction group and risperidone group; The schizophrenia model was established by intraperitoneal injection 0.1mg/kg MK801 for 2 weeks except the normal group,and then evaluated by open field test and water maze.Xianhuan Decoction group were given 12 g/kg Xianhuan Decoction and risperidone group were given 0.2 mg/kg risperidone for 4 weeks respectively, the other groups were treated with placebo.The open field test and electrochemical coulometric array HPLC were used to detect the relevant data after 4 weeks.Results The score of stereotyped behavior in model rats was (2.54 ± 0.52,2.42 ± 0.51,2.30 ± 0.48 ) ,it was higher than the normal group( 1.08 ± 0.28 ), the models were successful according the standard.After drug intervention, the locomotor activity of Xiaohuan Decoction and risperidone group (780.57 ± 248.99,819.76 ± 267.21 ) was less than the model group (1204.15 ± 344.25 ), there was a significant difference(P ＜ 0.05 ) ;and escape latency latency of Xiaohuan Decoction (30.25 ± 13.67) was shorter than that the model group (44.72 ± 19.88) ,the number of units to wear (5.67 ± 1.44) was more than that the model group ( 3.08 ± 1.44 ), there were significant difference (P＜ 0.05 ).The content of 5-HT in striatum were significantly decreased in Xianhuan Decoction group(0.08 ± 0.26) and risperidone group (0.11 ±0.07) compared with model group (0.25 ±0.14) (P ＜ 0.05 ) , and there was no significant difference of the content of 5-HIAA between Xianhuan Decoction group(0.28 ±0.36) and model group(0.27 ± 0.09 ) (P ＞ 0.05 ).Conclusion Locomotor activity and memory impairment were improved by Xianhuan Decoction, and the possible mechanism may be that Xianhuan Decoction can regulate the content of 5-HT in striatum in the schizophrenia model rat treated.
ABSTRACT
Objective To observe the results of multifocal visual evoked potential (mfVEP) examination in patients with anterior ischemic optic neuropathy (AION) before and ater treatment,and to probe its clinical significance.Methods A total of 90 patients (90 eyes) with AION were examined by mfVEP;the second-order reaction of mfVEP was analyzed.The reaction was divided into upper and lower hemi-field of visual field,or 1 / 4 quadrant visual field (superior nasal,inferior nasal,superior temporal,and inferior temporal).The sum of waves of each response was analyzed and the results in various regions were compared.The features of wave configuration was compared between the AION eyes and the contralateral eye,and between the AION eyes before and after treatment.Results The amplitude and latency of P-wave of mfVEP was 0.198±0.033 and 100.197±7.354 respectively in AION eyes before treatment,and was 0.271±0.024 and 98.567±6.794 in the contralateral eyes;the difference was significant (t=16.556,18.330;P＜0.01).The amplitude and latency of P-wave of mfVEP was 0.229±0.016 and 100.104±10.603 respectively in AION eyes after treatment,which differed much from that before the treatment (t=13.649,8.858;P＜0.01) and also from that of the contralateral eyes (t=13.649,8.858;P＜0.01).Conclusions The amplitude and latency of P-wave of mfVEP may accurately reflect the recovery of local optic nerve damage in AION eyes before and after treatment with good repeatability.AION can be used as a new method for AION diagnosis and detection of the prognosis.
ABSTRACT
We studied the ligation of coagulation factor VIII heavy and light chains in Escherichia coli by utilizing the intein-mediated protein trans-splicing. A B-domain deleted factor VIII (BDD-FVIII) gene was broken into two halves of heavy and light chains before Ser1657 which meets the splicing required conserved residue and then fused to 106 and 48 amino acid-containing N-part termed Int-N and C-part termed Int-C coding sequences of split mini Ssp DnaB intein respectively. These two fusion genes were constructed into a prokaryotic expression vector pBV220. Through induction for expression of recombinant protein it displayed an obvious protein band as predicted size of BDD-FVIII protein on SDS-PAGE gel. Western blotting using factor VIII specific antibodies confirmed that this protein band is BDD-FVIII produced by protein trans-splicing. It demonstrated that the heavy and light chains of BDD-FVIII can be efficiently ligated with the Ssp DnaB intein-mediated protein trans-splicing. These results provided evidence for encouraging our ongoing investigation with intein as a means in dual AAV vectors carrying the factor VIII gene to overcome the packaging size limitation of a single AAV vector in hemophilia A gene therapy.