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1.
Chinese Journal of School Health ; (12): 1086-1089, 2022.
Article in Chinese | WPRIM | ID: wpr-936544

ABSTRACT

Objective@#To construct and determine the weight of Index System for Assessing Parent s Ability on Child Injury Prevention, and to provide the basis for childhood injury intervention in family.@*Methods@#Twenty four experts majored in related fields were invited to participate in Delphi consultation. The final index system was constructed according to the consulting results and the weight of primary and secondary indicators were calculated.@*Results@#The final index system consisted of 5 subscales corresponding to 5 age groups: 0,1-2,3-5,6-11 and 12-17 years old. Each scale included 4 primary indicators and 11 secondary indicators. The weight of primary indicators obtained by analytic hierarchy process is 0.28 for "environment", 0.16 for "product", 0.31 for "behaviors and skills", and 0.25 for "psychology". The highest weight of secondary indicators for "environment", "product" and "behaviors and skills" was "water area", while the top secondary indicators for "psychology" included "parental style""emotional control" "family atmosphere", with all indicators weighted higher than 0.2.@*Conclusion@#The Index System for Assessing Parent s Ability on Child Injury Prevention by Delphi consultation is comprehensive in content, and with the focus on parental behaviors and skills on injury prevention.

2.
Chinese Journal of Practical Nursing ; (36): 612-616, 2020.
Article in Chinese | WPRIM | ID: wpr-864456

ABSTRACT

Objective:To summarize the nursing safety management measures of cataract surgery in the Caribbean in order to provide experience for medical assistance in the future.Methods:A retrospective study was conducted to analyze the safety factors of 344 patients (355 eyes) undergoing cataract surgery in the Caribbean. The safety management measures included: pre-investigation preparation, perfect preoperative examination, perioperative systemic disease nursing and health education, strict safety verification and operating room management.Results:On the day of operation, 2 patients were changed due to high blood pressure and blood glucose, 1 patient was rescheduled due to deviation in the understanding of surgical eyes, the completion rate of operation was 99.13% (341/344) , and 2 eyes of traumatic cataract were not implanted with intraocular lens (IOL). The success rate of operation was 99.44% (353/355) . One day after operation, corneal edema occurred in 63 eyes (17.75%) and intraocular lens shifted in 1 eye. During the follow-up period, all patients had no infective endophthalmitis, and there was no significant progress in patients with systemic disease.Conclusions:The working conditions of health care are difficult, the population of the Caribbean is aging, most of them are accompanied by serious systemic diseases, and the condition of patients is complex. Through a series of safety management measures, the operation can be carried out safely and successfully and good results can be achieved.

3.
Chinese Journal of Postgraduates of Medicine ; (36): 153-156,157, 2017.
Article in Chinese | WPRIM | ID: wpr-606164

ABSTRACT

Objective To explore the relationship between 24 h ambulatory blood pressure monitoring (24h-ABPM) parameters, circadian rhythm and urinary albumin to creatinine ratio (UACR), urineβ2 microglobulin (β2-MG) in elderly patients with hypertension. Methods One hundred and forty-eight patients with essential hypertension (≥60 years old) were included in this study. 24h-ABPM was performed, and nocturnal blood pressure decline rate (BPR) was calculated. The patients were divided into two groups according the BPR:dipper group with 40 cases and non-dipper group with 108 cases. The levels of 24h-ABPM parameters, UACR and β2-MG were compared between two groups, and the relationship between UACR and 24h-ABPM parameters were analyzed. Results The levels of 24hSBP, dSBP and 24hDBP in two groups had no significant differences (P>0.05). Compared with those in dipper group, the levels of nSBP, nDBP in non-dipper group were significantly increased, and the level of dDBP was significantly decreased, P0.05). The result of Sspearman analysis showed that UACR had significant correlation with 24hDBP, nDBP, 24hSBP, nSBP and 24hPP (P<0.05 or<0.01). The result of multivariate linear regression analysis showed that UACR was independently correlated with nSBP (P<0.05). Conclusions The abnormal circadian rhythm in elderly hypertensive patients is closely related to early renal damage.

4.
Basic & Clinical Medicine ; (12)2006.
Article in Chinese | WPRIM | ID: wpr-594197

ABSTRACT

Objective To investigate the difference of differentiation from rat bone marrow mesenchymal stem cells(MSCs) from normal group and hepatic fibrosis model group into hepatocyte-like cells.Methods Wistar rats were randomly divided into two groups: normal group and hepatic fibrosis model group.The liver fibrosis was induced by CCL4.MSCs were isolated by combining gradient density centrifugation with plastic adherence.Pure MSCs were obtained by cultivation and passage.The cells then treated with HGF and FGF-4.Levels of AFP and albumin from supernatant were determined on day 15,21 and 27.On day 27,cells of induced and non-induced were collected,glycogen store of hepatocytes and the expressions of CK-18 and CK-19 were detected.Results The level of AFP in induced MSCs was higher on day 15,21,27,and reached the peak on day 21;there was no significant difference between induced and non-induced MSCs in albumin levels on day 15,but on day 21,27,compared with the non-induced MSCs,the albumin level in the induced MSCs was higher and reaches its peak on day 27;glycogen storage of induced MSCs was measured on day 27 as compared with non-induced MSCs;the induced MSCs expressed CK-18 and CK-19 while the non-induced MSCs did not.Compared by the levels of AFP and albumin,there was no significant difference in differentiation effect of MSCs between the normal group and hepatic fibrosis model group.Conclusion Rat MSCs of hepatic fibrosis model group could differentiate into hepatocyte-like cells with hepatic phenotype and biological function in the presence HGF and FGF-4.

5.
Chinese Journal of Physical Medicine and Rehabilitation ; (12)2003.
Article in Chinese | WPRIM | ID: wpr-682372

ABSTRACT

Objective To investigate the clinical and electrophysiological characteristics of cubital tunnel syndrome (CTS). Methods The clinical and electrophysiological data of 150 cases of CTS involving 173 upper limbs (UL) were collected. And the electrophysiological data of 76 healthy subjects were also collected. The data of EMG between the two groups were compared and analyzed statistically. Results Fibrillation potentials were detected in 114 and 91 UL, respectively, in abductor digiti minimi, and positive sharp waves in 50 and 48 UL, respectively, in the first dorsal interosseous muscle. The average conduction velocity of the ulnar nerve was decreased, with motor conduction velocity(MCV) from above to below elbow 34.6?9.75 m/s and sensory conduction velocity (SCV) 45.99?9.65m/s; the motor latency was prolonged and amplitude of motor action potential decreased. There was statistical difference between the patients and the healthy control groups ( P

6.
Journal of Third Military Medical University ; (24)2003.
Article in Chinese | WPRIM | ID: wpr-556422

ABSTRACT

Objective To summarize the clinical, electrophysiological, pathological characteristics of chronic inflammatory demyelinating polyradiculoneuropathy (CIDP). Methods From Jan 1990 to Dec 2003, 32 patients of CIDP were admitted by our hospital, 14 male and 18 female. The age range of onset of illness was 13 to 74 years old (averaging 42.56) with the prime period of 40 to 50. Before hospitalization, the course was from 2 months to 5 years. Among all patients, there were 8 cases of a relapsing course and 24 cases of a chronic progressive course. Twenty-four patients were treated with corticosteroids, while 7 received immuglobin (IG) and corticosteroids. The clinical data before and after the treatment was studied retrospectively. Results Most of the patients had subacute or chronic onsets. The common initial symptoms are numbness, paresthesia, and extremity weakness. Diplopia, decreased visual acuity, dysarthria, and dysphagia could also be found initially. It was usually a symmetric sensorimotor neuropathy with either a relapsing course or a chronic progressive course. It could be accompanied with autonomic dysfunction and cranial nerve involvement. Electromyogram demonstrated that the motive and sensory nerve conduction velocities were slow. The sural nerve biopsy showed demyelination and remyelination. `IG and corticosteroids were both effective. Conclusion CIDP might result in widespread peripheral nerve damages, in which autonomic dysfunction and cranial nerve involvement were common. The dominant electrophysiological changes showed peripheral nerve demyelination accompanied by axon degeneration. The sural nerve biopsy played an important role of diagnosis. The treatment with IG and corticosteroids was a most effective way for CIDP nowadays.

7.
Chinese Medical Journal ; (24): 1465-1469, 2002.
Article in English | WPRIM | ID: wpr-282163

ABSTRACT

<p><b>OBJECTIVE</b>To perform cloning of the gene encoding Chinese Schistosoma japonicum tropomyosin (SjcTM) and its expression in Escherichia coli.</p><p><b>METHODS</b>SjcTM cDNA fragment, except for 14 amino acids at the amino terminus, was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) with total RNA extracted from adult worms of S. japonicum. The RT-PCR product was cloned into T vector and sequenced. The SjcTM cDNA, derived from the constructed TA clone pGEM-SjcTM, was then subcloned into the expressing vector pBV220. After characterization by agarose gel electrophoresis, endonucleases digestion and PCR, the resultant recombinant plasmid was used for expression under the temperature-dependent condition.</p><p><b>RESULTS</b>The RT-PCR product, cloned into a T vector, was sequenced and shown to be 96.5% identical at the nuclei acid level and 98.1% identical in deduced amino acid sequence to that of S. mansoni tropomyosin. The target DNA fragment was then subcloned into a prokaryotic vector pBV220. Induced expression in E. coli DH5alpha cells resulted in a constant level of recombinant protein production. The results of SDS-PAGE and Western blot revealed that the molecular weight of non-fusion recombinant protein (rSjcTM) was approximately 32 kDa and could be recognized specifically by a polyclonal antiserum specific for native S. japonicum tropomyosin (SjcTM).</p><p><b>CONCLUSION</b>The engineering of the cDNA encoding S. japonicum tropomyosin and its bacterial expression was successfully made.</p>


Subject(s)
Animals , Amino Acid Sequence , Base Sequence , Cloning, Molecular , DNA, Complementary , Chemistry , Escherichia coli , Genetics , Molecular Sequence Data , Recombinant Proteins , Schistosoma japonicum , Genetics , Tropomyosin , Chemistry , Genetics
8.
Chinese Journal of Nervous and Mental Diseases ; (12): 4-9, 2001.
Article in Chinese | WPRIM | ID: wpr-411961

ABSTRACT

Objective  To study the clinical and electrophysiological features in Charcot-Marie-Tooth disease type 1A with gene duplication.Methods Clinical symptoms and signs were summarized in 22 patients from 21 unrelated families. Electromyography (EMG) as well as motor conduction velocities (MCV) and sensory conduction velocities (SCV) examinations were performed in all patients. Results Evidence of CMT was initially detected within the second decade in 18 patients. Nearly half of patients were sporadic cases. The typical clinical manifestations of CMT1A were weakness and atrophy in the distal limbs, weakness or absence of the tendon reflexes, talipes equinovarus and postural tremor the upper limb. Additionally, some special symptoms and signs were also observed occasionally, including brisk tendon reflexes, extensor plantar responses, scoliosis, foot ulcers and nystagmus. EMG revealed that 77.3% of the patients had fibrillation and positive sharp potentials. 81.8% of them had prolonged motor unit potential limit. Median MCV showed there was no significant difference between CMT1A patients and CMT1 patients without duplication (t=1.63, P>0.05). Values of SCV and MCV for the lower limbs were not obtained in 20 patients and more than 2/3 of the patients respectively. Conclusions The clinical features of CMT1A included high frequent of sporadic cases, early onset in the second decade and various manifestations. The electrophysiological features were that the damages of nerves for the lower limbs were more severer than those in the upper limbs and the damages of the sensory nerves were more severer than those of the motor nerves. The phenotype was variable although the genotype was the same in CMT1A patients with PMP22 duplication.

9.
Chinese Journal of Parasitology and Parasitic Diseases ; (6)1997.
Article in Chinese | WPRIM | ID: wpr-683931

ABSTRACT

ObjectiveTo observe the dynamics of antibodies and protection against Schistosoma japonicum infections in buffaloes after immunized with recombinant 26 kDa glutathione S transferase (reSjc26GST). Methods Buffaloes in 2 villages endemic for schistosomiasis japonica were selected as test and control groups, respectively.In test group initially 96 buffaloes were vaccinated with reSjc26GST, and 90 buffaloes in the control group did not experience vaccination. The indicators included levels of antibodies to reSjc26GST in buffaloes before and after infection with S japonicum and changes in infection rate. Results Specific antibodies, which showed a trend of trapezoid increase, were induced in buffaloes after immunized with reSjc26GST. Twenty months after immunization, the infection rate of the test group was decreased by 62 2% when compared with that before vaccination,and by 67 7% when compared with that of the control in the corresponding period.Conclusion Specific antibodies and a certain extent of protection were induced in buffaloes after immunized with reSjc26GST, which played an significant role in ameliorating morbidity.

10.
Chinese Journal of Schistosomiasis Control ; (6)1989.
Article in Chinese | WPRIM | ID: wpr-564040

ABSTRACT

Objective To obtain novel vaccine candidate antigens against Schistosoma japonicum. Methods S. japonicum schistosomula cDNA library was screened by using sera of Microtus fortis that was naturally resistant to schistosomiasis. The positive clones were transformated into Escherichia coli BM25.8, E. coli clones containing the plasmid cultured in LB, and then selected for plasmid extraction, the plasmid DNA was digested by EcoRⅠand Hind Ⅲ, and analysed by agarose gel electrophoresis. The positive clones were also sequenced and the data were analysed through the internet Nucleotide BLAST software of NCBI and Expert Protein Analysis system of GeneRunner and HNN. Results Twelve positive clones were obtained after repeatedly immunoscreening the library and their sizes ranged from 300 bp to 1100 bp. Two novel genes (named as Sj-sMf1 and Sj-sMf2) with complete ORF were obtained. The deduced protein of Sj-sMf1 consisted of 93 amino acids while Sj-sMf2 consisted of 61 amino acids. Sj-sMf1 protein predicted containing one cAMP phosphorylation site and Casein kinase C phosphorylation site, respectively. Sj-sMf1 protein predicted containing one Casein kinase C phosphorylation site and two Protein kinase C phosphorylation sites. Conclusion Two novel genes predictably encoding unknown proteins are obtained from immunoscreening of Schistosoma japonicum schistosomula cDNA library by M. fortis sera.

11.
Chinese Journal of Schistosomiasis Control ; (6)1989.
Article in Chinese | WPRIM | ID: wpr-561993

ABSTRACT

Objective To clone and express the gene encoding Schistosoma japonicum myophilin-like protein (SjcMLP) and to study the antigenicity of the recombinant protein. Methods The SjcMLP gene was amplified by PCR. The PCR product was cloned into T vector, and then subcloned into expression vector pQE30. The recombinant plasmid of pQE30-SjcMLP was transformed into E.coli M15, and induced with IPTG for expression. The bacterial lysis was conducted by ultrasonication and the supernatant was analysed by SDS-PAGE. The recombinant protein (reSjcMLP)was purified with the Ni-NTA resin, and analysed with SDS-PAGE and Western blot. The titers of sera from C57BL/6 mice immunized subcutaneously with reSjcMLP were detected by ELISA. Results The results of SDS-PAGE and Western blot showed that the molecular weight of expressed fusion protein was around 24.8 kDa and was recognized by the sera from the mice infected with Schistosoma japonicum. The purified protein of reSjcMLP was coated for ELISA test and the IgG titers in the sera from the mice immunized with reSjcMLP were as high as 1∶12 800 reacted with. However, no significant difference was found in worm reduction rates between the immunized mice and control mice. Conclusions The fused recombinant protein of reSjcMLP is successfully ex-pressed and purified. The recombinant protein in this experiment fails to induce significant protection against the challenge infection in C57BL/6 mice.

12.
Chinese Journal of Schistosomiasis Control ; (6)1989.
Article in Chinese | WPRIM | ID: wpr-561611

ABSTRACT

Objective To perform the cloning of the gene encoding Schistosoma japonicum Chinese-strain hypoxanthine-guanine phosphoribosyltransferase(HGPRT)and its expression in Escherichia coli.MethodsA couple of primers were designed with the BamHI restriction endonuclease site introduced in forward primer and SalI in reverse primer.Total RNA was isolated from adult worms of S.japonicum Chinese-strain(Anhui-strain,Sjc-A)and the SjcHGPRT gene was amplified by reverse transcriptase-polymerase chain reaction(RT-PCR).The PCR product and the prokaryotic expression vector pET28a were digested by both restriction endonucleases BamHI and SalI.The target DNA fragments were purified and cloned properly into pET28a.After identification by en-donucleases digestion,PCR and sequencing,the recombinant plasmid pET28a-SjcHG PRT was transformed into competent E.coli BL21 and expressed in the presence of IPTG.Results pET28a-SjHGPRT was sequenced and shown to be 99% and 83% identical in deduced amino acid sequence to that of S.japonicum Chinese-strain(Hunan-strain,Sjc-H)and S.mansoni HGPRT,respectively.The results of SDS-PAGE and Western blot revealed that the molecular weight of expressed protein was around 30 kDa and could be recognized by anti-His-G-HPR antibody and sera from mice and human with schistosomasis japonica.Conclusion The recombinant plasmid containing SjcHGPRT cDNA is successfully constructed and its expression protein(reSjcHGPRT)is also successfully purified.

13.
Chinese Journal of Schistosomiasis Control ; (6)1989.
Article in Chinese | WPRIM | ID: wpr-557043

ABSTRACT

ObjectiveTo construct and identify a DNA vaccine of Schistosoma japonicum (Chinese strain) thioredoxin. MethodsAccording to a cDNA sequence of S.japonicum (Philippine strain) thioredoxin, a couple of primers was designed with the BamH I restriction endonuclease site introduced in forward primer with ATG as start condon and EcoR I in reverse primer with TCA as termination codon. The gene encoding S.japonicum (Chinese strain) thioredoxin (SjcTrx) was amplified by RT-PCR using total RNA of schistosome as the template. The PCR products and the pcDNA3 plasmids were digested by restriction endonucleases BamH I and EcoR I. The target DNA fragment were purified and cloned properly into the eukaryotic expression vector of pcDNA3. The recombinant plasmids were then transformed into competent E.coli JM109 and identified by endonucleases digestion, PCR, agarose gel electrophoresis and sequencing. ResultsThe RT-PCR product was around 334 bp judged by agarose gel electrophoresis. The same fragments were obtained by restriction enzyme digestion from the recombinant plasmid and PCR with the plasmid DNA as a template. The recombinant plasmid, designated pcDNA3-SjcTrx, was sequenced and shown to be 97% and 43% identical in deduced amino acid sequences to that of S.japonicum (Philippine strain) and S.mansoni thioredoxin, respectively. ConclusionThe S.japonicum thioredoxin DNA vaccine had been constructed successfully, and further studies will be made in different animal models for its immunogenicity.

14.
Chinese Journal of Parasitology and Parasitic Diseases ; (6)1987.
Article in Chinese | WPRIM | ID: wpr-683838

ABSTRACT

Objective To clone and express the cDNA encoding Schistosoma japonicum tropomyosin. Methods The cDNA was amplified by reverse transcription polymerase chain reaction (RT PCR). The PCR products were ligated with pGEM T vectors and then for transformations. After characterization of white clones by agarose gel electrophoresis, endonucleases digestion and PCR, some recombinant plasmids with inserts were used for sequencing. Then the gene was subcloned into prokaryotic expression vector pQE30 and expression was induced by IPTG. Results The PCR products was 823 bp judged by agarose gel electrophoresis and sequencing. A cDNA encoding S japonicum tropomyosin, except for 14 amino acids at the amino terminus and 2 at the carboxyl terminus, has been constructed and cloned successfully. The colony, designated pGSjcTM12, was sequenced and shown to be 91 1% identical at the nuclei acid level and 98.1% identical in deduced amino acid sequence to that of S mansoni tropomyosin. The gene was subcloned into pQE30 and an expressed protein of about 32 kDa was obtained.Conclusion The cloning and expression of the gene encoding S japonicum tropomyosin had been successfully made.

15.
Chinese Journal of Parasitology and Parasitic Diseases ; (6)1987.
Article in Chinese | WPRIM | ID: wpr-589382

ABSTRACT

Objective To clone and express the partial encoding sequence of Mr 70 000 heat shock protein of Cryptosporidium andersoni (CaHSP70) in Escherichia coli and identify the recombinant protein. Methods Total RNA was extracted from oocysts of C.andersoni isolated from Xuzhou, Jiangsu (XZ-BOV). The CaHSP70 gene was amplified by RT-PCR. The PCR product was cloned and then subcloned into pET28a vector, and the recombinant plasmids were transformed into E.coli BL21(DE3) subsequently. The expressed protein induced by IPTG was purified and identified by SDS-PAGE and Western blotting, and was further analyzed by relevant bioinformatics softwares. The specific IgG antibodies in mice immunized by rCaHSP70 were detected by Western blotting and ELISA respectively. Results The deduced amino acid sequence showed to be identical with that of C. andersoni Mr 70 000 heat shock protein (HSP70). The recombinant protein expressed in the form of inclusion body was about Mr 43 000. It could be recognized by anti-His G labeled HRP antibodies and all the sera from mice infected with C. andersoni and children infected with C. parvum as well as sera from mice immunized with rCaHSP70 respectively. The rCaHSP70 possibly had multiple domains and potential antigenic determinants. Phylogenetic analysis showed that XZ-BOV and C. andersoni were in the same clade. ELISA showed that the level of specific antibodies against rCaHSP70 in immunized BALB/c and C57BL/6 mice was significantly higher than that of mice before immunization. Conclusion The recombinant plasmid pET28a-CaHSP70 has been constructed. The purified rCaHSP70 exhibits high antigenicity and seems a potential candidate antigen for immunodiagnosis of cryptosporidiosis.

16.
Chinese Journal of Parasitology and Parasitic Diseases ; (6)1987.
Article in Chinese | WPRIM | ID: wpr-588948

ABSTRACT

Objective To isolate and identify Cryptosporidium oocysts from feces of naturally infected cow. Methods Fecal samples were collected from Cryptosporidium infected cows confirmed by modified acid-fast staining method. Oocysts were isolated and purified with Sheather sucrose density gradient centrifugation technique. Genomic DNA was isolated with Chelex-100. Both primers were designed to amplify Cryptosporidium small subunit ribosome RNA gene (SSU rRNA) and Cryptosporidium oocyst wall protein gene (COWP), respectively. The PCR products were cloned into pGEM-T and pGEM-T Easy vector and sequenced subsequently. Homology and phylogeny were analyzed with BLASTn and MEGA software. Results The results suggested that the size of oocysts was (7.4?0.32)?m by(5.4?0.21)?m and the ratio of length and width was 1.37?0.07 (n=20). BLASTn revealed that the identity of SSU rRNA and COWP gene of Cryptosporidium isolated from cow to the counterparts of C.andersoni was 100% and 99% respectively. Phylogenetic reconstruction placed the isolated Cryptosporidium within the C.andersoni clade based on the sequence of SSU rRNA and COWP gene. Conclusion What isolated from naturally infected cow feces has been identified as C. andersoni.

17.
Chinese Journal of Parasitology and Parasitic Diseases ; (6)1987.
Article in Chinese | WPRIM | ID: wpr-586851

ABSTRACT

Objective To investigate the protective immunity of the recombinant thioredoxin of Schistosoma japonicum(reSjcTrx)in mice. Methods Thirty 6-week old female C57BL/6 mice were randomly divided into 3 groups with 10 each: reSjcTrx with Montanide ISA720 adjuvant, adjuvant control, and infection control. Mice were vaccinated subcutaneously at week 0, 2, 4 with reSjcTrx emulsified in Montanide ISA720 adjuvant. The mice in adjuvant group was injected three times with Montanide ISA720 and saline only. Mice in infection control group were given no injection. Three weeks after final injection, each mouse was challenged with 30?1 cercariae of S. japonicum (Chinese strain). At the week six after challenge, all mice were sacrificed and perfused. The number of recovered worms and eggs from liver tissue of mice were counted. Sera were collected from mice before immunization, before challenge and before killing. The anti-SjcTrx antibodies in sera were detected with ELISA. Results ELISA showed a high level of specific IgG antibodies in mice immunized with the reSjcTrx. The worm reduction rate and egg reduction rate of reSjcTrx immunization group were 22.8% and 29.5% respectively, significantly higher than those of the control groups (P﹤0.05). SDS-PAGE and Western blotting revealed that the molecular weight of expressed protein was around Mr 14 000 and could be recognized by sera from rabbit infected with S.japonicum and from mice immunized with reSjcTrx. Conclusion The reSjcTrx induces certain protective immunity against schistosomiasis japonica in mice.

18.
Chinese Journal of Parasitology and Parasitic Diseases ; (6)1987.
Article in Chinese | WPRIM | ID: wpr-585144

ABSTRACT

Objective To establish three methods of DNA extraction from Cryptosporidium parvum oocysts and test by PCR. Methods After three freeze-thaw cycles, three kinds of templates were extracted from the oocysts by Chelex-100, phenol/chloroform or genomic DNA purification system kit, and used for PCR detection. According to the sequence of a C.parvum gene (L16996), a pair of primers was designed and synthesized, and used for PCR. The sensitivity of the template by Chelex-100 method was also tested by PCR. Results One 446 bp PCR product was observed by agarose gel electrophoresis for all three kinds of templates. The PCR sensitivity by Chelex-100 extracted DNA reached for detection of a specimen containing only 1/2 oocyst. Conclusion The three kinds of extraction can all be served as templates for PCR detection of C.parvum oocysts, while Chelex-100 method is simpler, quicker and more reliable for DNA extraction of the parasite.

19.
Chinese Journal of Parasitology and Parasitic Diseases ; (6)1987.
Article in Chinese | WPRIM | ID: wpr-582130

ABSTRACT

Objective To understand and identify the molecules related to the natural resistance to Schistosoma japonicum infection in Mirotus fortis. Methods Sera from Mirotus fortis without schistosome infection were collected. The S.japonicum adult worm cDNA library was immunologically screened with the sera. The positive recombinants were identified, cloned, sequenced and analysed with software and internet. Results Seven genes encoding antigens relevant to sera antibodies in Mirotus fortis were cloned and sequenced. These antigens included glyceraldehyde 3-phosphate dehydrogenase (GAPDH), serine protease inhibitors(SERPIN), 70 kDa heat shock protein(HSP70), 22^6 kDa membrane-associated antigen, paramyosin (Sj97), cytochrome C and cathepsin B. Conclusion Many protein molecules might have been involved in natural resistance to \{S.japonicum\} infection in Mirotus fortis. The above 7 kinds of molecules may be identified as new candidates of vaccine against \{S.japonicum\} infection.

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