Monascus spp.,a kind of filamentous fungi,produce abundant of important metabolites which were widely used in the fields of food and medicine.Until now,there are few reports on the important functional genes of the Monascus spp.due to little genetic information.In this paper,the feasibility of gene deletion mediated via Agrobacterium tumefaciens on the basis of homologous recombination was analyzed by studying on the deletion of the RGS domain of putative G-protein signaling regulator gene mrfA in Monascus ruber.The length of homologous arms of deletion vector pC805S were 958 bp and 824 bp,respectively.There were 26 transformants in which homologous recombination occurred in 138 transformants and the recombination rate was 18.8%.The result showed it was feasible to identify the function of unknown gene in M.ruber with the targeted-deletion technology mediated via A.tumefaciens.

To study the clinical skills training methods and formulate a training objective for pediatric long-year-program students, to let the students, through three years of training program, reach the clinical ability as a chief resident, with better laboratory research skills, teaching awareness and responsibility, and also with good comprehensive quality and excellent English, and finally become clinical talents with great potential and innovation, this report summarized our study results of training two terms of long-year-program pediatric graduates, who fully meeted the training objectives in the department's practice.

Objective To evaluate the effects of the chronic Keshan disease's self-management in Liangshan Prefecture of Sichuan Province. Methods According to "The Serf Management Program of Chronic Keshan Disease in Liangshen Prefecture of Sichuan Province", 56 chronic Keshan disease patients were selected in the personalized self-manngemant evaluation under the instructions by endemic disease specialists and the rural doctors. Evaluation was based on changes of indexes such as the clinical symptoms, general health conditions, the electrocardiogram, X-ray, the heart function, etc, before treatment and 3 and 6 months following the treatment. Results Clinical signs and symptoms of the patients were significantly improved 3 and 6 months after the treatment, and the improvement was more obvious 6 months than 3 months following the treatment(P<0.05 or< 0.01). After treatment for 3 months, the patients'electrocardingram and heart function did not show obvious change (X2=0.05,039, P0.05); hut obvious improvements Eexcept X ray results(X2=0.61 ,P0.05)] were seen 6 months after treatment (X2=4.36,16.84, P<0.05 or<0.01). Altogether, among the 56 patients evaluated after treatment for 6 months, none achieved the clinical cure standard, 26 cases(46.3%) showed significant improvement, 17 cases (30.4%) were stable, 5 cases (8.9%) were aggravated,one case (1.8%) lost contact, and 6 cases (10.6%) died. Conclusion The project of the chronic Keshan disease's self-management is suitable for the present situati,on of the endemic regions and can he introduced to many places in our country.

To identify SJCHGC01743 gene of Schistosoma japonicum and evaluate the potential of the recombinant protein as a new vaccine candidate for schistosomiasis, polymerase chain reaction (PCR) technique was used to amplify the cDNA of the gene and real-time RT-PCR was used to analyze the transcription profiles of SJCHGC01743 at different development stages. Recombinant plasmid was successfully constructed and transformed into competent Escherichia coli BL21 (DE3). Then the recombinant protein was expressed, purified and emulsified with ISA206 adjuvant to immunize BALB/c mice for three times. The immunogenicity was confirmed by Western blotting and tissue localization was detected by indirect immunofluorescent assay. The specific antibody level was detected by ELISA. The immunoprotection of rSjOST48 was evaluated by the reduction in worm and egg counts in mice. A cDNA with 1 248 nucleotides was isolated from 28-day-old schistosomes cDNAs by PCR. Sequence analysis revealed that SJCHGC01743 was a 48-kDa subunit of the oligosaccharyltransferase complex (OST48) and named as SjOST48. Real-time PCR analysis indicated that this gene was expressed in all investigated stages and had the highest expression level in 28 d worms, the level of gene transcription in female worms was significantly higher than that of male worms. Then recombinant plasmid pET28a(+)-SjOST48 was successfully constructed and expressed in E. coli BL21 (DE3). Western blotting analysis showed that rSjOST48 had good immunogenicity. Indirect immunofluorescent analysis revealed that SjOST48 was mainly distributed on the tegument of the worms. The result of ELISA indicated that the rSjOST48 vaccinated group could induce a significant increase in the level of specific IgG, IgG1 and IgG2a. An immunoprotection experiment showed that the vaccination of rSjOST48 in mice induced 32.62% (P < 0.05) reduction in the numbers of worms and 57.61% (P < 0.01) in eggs in liver, compared with that of the control group. This study provides the foundation for proceeding further research on the biological function of SjOST48 and screening new vaccine candidates for schistosomiasis.
Animals ; Antibodies, Helminth ; blood ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Female ; Genes, Helminth ; Helminth Proteins ; genetics ; immunology ; Immunoglobulin G ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Real-Time Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; Schistosoma japonicum ; genetics ; Schistosomiasis japonica ; prevention & control ; Vaccination

Objective To investigate the effects of a novel |3 titanium fixation with high strength and low elastic modulus on bone healing of osteoporotic fracture and its effects on stress shielding.Methods Forty female Sprague-Dawley rats were oophorectomized before establishment of femur fracture model.The fracture of left femur was stabilized by titanium alloy pin with high elastic modulus (110 Gpa,high elasticity modulus group) and the fracture of right femur by titanium alloy pin with low elastic modu-lus (30 Gpa, low elasticity modulus group).Six and 12 weeks later, the bone volume (BV) , total vol-ume (TV) and bone volume fraction (BVF) were detected by Micro-CT, and the maximum load was de-tected by biomechanical test.The effects of two titanium alloy pins on formation of bony calluses and me-chanical properties of bones were compared.Results Compared with the low elasticity modulus group, more collagen and bony callus, thicker bone trabeculae and more regular lamellar bone were found in the high elasticity modulus group.In low elasticity modulus group, TV (ROI1) was increased obvious-ly at week 6, with no statistical differences in BV (ROI 1), BV (ROI 2) and maximum load between two groups (P < 0.05).TV and BV in region ROI 1 in the low elasticity modulus group were significantly higher than that in high elasticity modulus group (P < 0.05) , while BV in region ROI 2 in low elasticity modulus group was significantly higher than that in high elasticity modulus group at week 12 postoperative-ly (P<0.05).Biomechnical test indicated that the maximum load in low elasticity modulus group was higher than that in high elasticity modulus group.Conclusions The novel B titanium alloy with high strength and low elastic modulus helps reduce effects of stress shielding, diminish bone resorption, in-crease the fibrous callus formation at the initial phase and improve the quality of bone healing at the later phase.

OBJECTIVETo study the infection status of Leptospira in rodents on Heixiazi island Heilongjiang province in 2011.

METHODSA total of 356 rodents were captured by night trap on the Heixiazi island from April to October 2011. The kidney tissue samples were collected by asepsis operation and the genomic DNA were extracted from them. Leptospira strains were confirmed by polymerase chain reaction(PCR) amplification of the 482 bp 23 S rDNA gene. Fifteen PCR products selected by the month were purified and sequenced by the methods of Sanger dideoxy, the sequences then compared with other Leptospira strains in Genebank, and phylogenetic analyses were drafted by software Mega 4.0.

RESULTSAmong 356 rodents, the dominant species were Clethrionomys rutilus (39.3%, 140/356) and Apodemus agrarius (36.0%, 128/356). The infection rate of Leptospira was 11.0%, with 39 rodent samples detected positive. All the rodent species were infected except for Rattus norvegicus. The infection rate was 9.4% (12/128) in Apodemus agrarius, 12.9%(18/140) in Clethrionomys rutilus, 10.8%(7/65) in Microtus fortis Buchner. No significant difference was found between the infection rate and the species of rodents by chi square test(χ(2) = 1.92, P > 0.05). Among months, the infection rate was 5.6% (4/72) in May, 8.8% (5/57) in June, 12.8% (5/39) in July, 9.8% (5/51) in August, 33.3% (11/33) in September, 22.5% (9/40) in October,but no infection in April. There was significant difference in infection in different months (χ(2) = 32.92, P < 0.05). All the Leptospira in rodents on the Heixiazi island were in the same phylogenetic branch with a high similarity of 97.1%-99.6%, close with the Australia strain U90865 by the similarity above 96.3%.

CONCLUSIONLeptospira is probably prevalent in rodents on the Heixiazi island, and the phylogene of the strains were similar. The infection rate in rodents was significantly different in months but not in hosts.

Animals ; China ; Leptospira ; isolation & purification ; Leptospirosis ; prevention & control ; Murinae ; microbiology ; Phylogeny ; Rats

Objective To establish a dual RT-PCR detection method for bovine viral diarrhea virus(BVDV)in bovine-derived samples. Methods The primers were designed and synthesized according to the published BVDV1 and BVDV2 genes containing highly conservative sequences in the 5' untranslated regions(5' UTR)to establish the dual RT-PCR method. The specificity,sensitivity,stability of this method were evaluated. Then 41 bovine-derived samples and 64 bovine plasma samples including bovine calf serum, deproteinized calf serum extract and one lienal polypeptide injection were detected with this method. Results There was no cross reaction with bovine parainfluenza virus type 3(BPIV3), classical swine fever virus(CSFV)and Japanese encephalitis virus(JEV)when samples were detected with the established dual RT-PCR method. The lowest concentration of template DNA for detection of BVDV1 and BVDV2 was 8.87 × 102copies and 6.31 × 102copies per microliter,respectively. Electrophoresis bands of about 151 bp and 303 bp were still amplified and detected after the BVDV1 and BVDV2 cDNA was stored at -30℃ for 12 months. The BVDV positive rate of 41 bovine-derived samples and 64 bovine plasma samples detected with this dual RT-PCR method was 14.6% and 29.7%, respectively. Conclusions The established dual RT-PCR method has the advantages of high efficiency, specificity,sensitivity and stability,and can be used for the detection of BVDV in bovine-derived samples.

Objective To investigate the diagnostic value of random forest(RF)model based on CT images and clinical data for preoperative T staging of colorectal cancer. Methods Four hundred and fifty patients with colorectal cancer who were pathologically confirmed by surgery and underwent preoperative CT examinationinthe first hospital of Jilin university from January 2016 to July 2016 were included retrospectively(Stage≤T2,T3,and T4 each has 150 cases).According to the ratio of 2:1,the patients were divided into training set(300 cases)and test set(150 cases,stage ≤T2,T3,and T4 each has 50 cases)by computer random software. Each of 450 patients had one lesion. All the patients underwent preoperative abdominal and pelvic contrast-enhanced CT scan.The clinical,imaging and pathological data[gender,age, carcinoembryonic antigen (CEA) level, carbohydrate antigen 19-9 (CA19-9) expression, intestinal wall deformation, maximum diameter of tumorand thickness of intestinal wall, location, enhancement homogeneity and enhancement rate]of these patients were collected.The correlation between the collected factors and pathological T staging was analyzed by Spearman correlation analysis.The preoperative staging model of colorectal cancer was established by RF algorithm in the training set.Two kinds of methods(model and traditional method)were used to diagnose T stage of the patients in the test set.The accuracy of the two methods was calculated by postoperative pathological staging as the gold standard.The consistency test was used to evaluate the consistency of the RF model results with the pathological results. Results T-staging was positively correlated with CEA, CA19-9, intestinal wall deformation, tumor size and thickness of intestinal wall(r=0.449,0.291,0.624,0.573,0.386;P<0.05).Age,location,enhancement homogeneity and enhancement rate were slightly negatively correlated with T-staging(r=-0.115,-0.245,-0.120 and-0.339;P<0.05).The predictive results of the model in≤T2,T3,and T4 stage cancers were moderately and highly consistent with the standard of pathology,and the Kappa value were 0.769,0.615 and 0.800,respectively.The total accuracy rate of the model andthe traditional method are 80.7%(121/150)and 54.0%(81/150). Conclusion Application of random forest model based on multi-slice spiral CT images and clinical data can improve the diagnostic efficacy of preoperative T stage of colorectal cancer.

OBJECTIVETo establish a new manufacturing method for Bletilla striata synthetic seeds, and provided a new way for rapid propagation of B. striata, the correlated influential factors were studied.

METHODThe synthetic seeds were manufactured by taking seeds of B. striata as materials which were beforehand germinated in 1/2 MS medium for 10 days, and the influential factors such as artificial endosperm components, episperm substances, storage conditions and germination groundmass impact on the germination rate and seedling rate of the synthetic seeds were evaluated.

RESULTCompound 4.0% sodium alginate + 0.2 mol x L(-1) CaCl2 + 0.4 mg x L(-1) penicillin + 0.3% carbendazim powder + 0.2% sodium benzoate served as the best episperm substances while MS + 1.0 mg x L(-1) NAA + 2.0 mg x L(-1) KT as the best endosperm components, in which, high germination rate and seedling rate were obtained. The synthetic seeds storing in the 4 degrees C for a long time was able to have still high vitality.

CONCLUSIONThe B. striata synthetic seeds manufacturing system was established successfully, while efforts should be taken to improve the sowing technique of the synthetic seeds in non-sterile conditions.

Cell Culture Techniques ; methods ; Culture Media ; chemistry ; metabolism ; Germination ; Orchidaceae ; growth & development ; metabolism ; Seedlings ; growth & development ; metabolism ; Seeds ; growth & development ; metabolism

Objective To evaluate the clinical efficacy of "Z"type aortic sinus angioplasty in acute Stanford A aortic dissection with severe aortic valve insufficiency .Methods A total of 384 cases of acute Stanford A aortic dissection were ana-lyzed retrospectively from January 2014 to March 2018 at Wuhan Asian Heart Hospital Great Vascular Center.There were 56 cases with severe aortic valve insufficiency, with 40 males and 16 females, aged 32-72 years old, averaged(53.92 ±9.26) years old.The root treatment of the 56 patients were all simulated with the "Z"type aortic sinus angioplasty technique, with 3 cases of distal semi-aortic arch replacement and 53 cases of total aortic arch replacement(placed with "trunk"stent).Results The average cardiopulmonary bypass time of all patients was(175.0 ±43.5)min, cross-clamp time was(133.3 ±31.9)min, and circulatory arrest time was(26.9 ±2.7)min.The degree of aortic valve regurgitation was 0-Ⅰin all patients after surger-y.Patients were followed-up 3 months to 4 years, the degree of aortic valve regurgitation of 45 cases was 0, 8 cases wasⅠ, and 3 cases wasⅡ.After operation, transesophageal ultrasonography immediately examined 2 cases with residual dissection of aor-tic sinus.Followed-up 1.5 years and 2.0 years respectively, no aneurysm was formed, and the diameter of the sinus of the aor-ta was 4.2 cm and 4.5 cm, respectively.No reoperation due to aortic valve regurgitation and sinus conditions was found during the follow-up.Conclusion The "Z"type aortic sinus angioplasty technique has a good clinical effect in acute Stanford A aor-tic dissection with severe aortic valve insufficiency.The operation is simple, safe and effective, easy to grasp and develop.