A temperature-sensitive mutant strain was isolated after transposon mutagenesis with Tn5 and named MT54.PCR was carried out with primers designed according to the sequence of transposon,the PCR products showed that the MT54 carried transposon in the genome. The mutant grew well at 30℃in minimal medium(MM)containing p-nitrophenol(PNP)as sole carbon source,while it cannot grow at 37℃in the same medium,NO_2.detection results also proved that.Comparing the degradation rate of PNP and hydroquinone of MT54 and DLL-E4 at different temperature,it was speculated that the mutant site locate in the PNP degradation related genes.

OBJECTIVETo evaluate the clinical effectiveness of Yiqi Huoxue Tongyang Xiezhuo Recipe (YHTXR, capable of supplementing qi, activating blood, warming yang, and discharge turbidity) in treating coronary atherosclerotic heart disease (CAHD). and chronic heart failure (CHF) with carotid plaque patients, and to explore new ways of Chinese medicine (CM).

METHODSTotally 69 CAHD-CHF patients of qi deficiency phlegm stasis syndrome (QDPSS) with carotid plaque were recruited in this study using parallel cohort method. They were assigned to the treatment group (35 cases) and the control group (34 cases). Patients in the control group received routine treatment of Western medicine, while those in the treatment group were additionally treated with YHTXR (twice daily). The therapeutic course for all was three months. Cardiac function levels, echocardiography, carotid plaque, blood lipids and safety indicators were observed before and after treatment.

RESULTSAfter treatment the improvement of cardiac function levels was better in the treatment group than in the control group (P < 0.05). Decreased LDL-C levels were higher in the treatment group than in the control group (P < 0.01). There was statistical difference in left ventricular ejection fraction (LVEF), carotid intima-media thickness (IMT), LDL-C, TC, TG in the treatment group between before and after treatment (P < 0.05). LDL-C and TG also decreased in the control group after treatment (P <0.05). There was no significant difference in the left ventricular ejection fraction, carotid IMT, or TC in the control group between before and after treatment (P > 0.05). There was no significant difference in stroke volume, left ventricular end-diastolic diameter, the area of carotid artery plaque, or HDL-C in the two groups between before and after treatment (P > 0.05).

CONCLUSIONSYHTXR could effectively improve cardiac functions of CAHD-CHF patients of QDPSS with carotid plaque, reduce blood lipids and IMT. It had no significant adverse reactions for elderly patients in short term.

Carotid Intima-Media Thickness ; Coronary Disease ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Heart ; Heart Failure ; drug therapy ; Humans ; Lipids ; Plaque, Atherosclerotic ; drug therapy ; Qi ; Ventricular Function, Left

OBJECTIVETo observe the effect of Wenyang Yiqi Huoxue Recipe (WYHR) on the apoptosis of photoreceptor cells in retinal degeneration slow (RDS) mice, and to investigate its molecular mechanisms.

METHODSRDS mice were randomly divided into the model group and the Chinese medicine group,and C57BL/6J mice were selected as the normal control group. Each group consisted of 4 female mice and 2 male mice. Mice in the Chinese medicine group were administered with WYHR (10 mg/g) by gastrogavage since mating. Baby mice drunk WYHR decoction instead of drinking water once they were born. The offspring were administrated with low dose WYHR decoction by gastrogavage from the 7th postnatal day, and the dose was increased to that for adult mice from the 21st postnatal day. Physiological saline was administrated to mice in the model group and the normal control group by gastrogavage. At 18, 28 and 48 postnatal days, electroretinogram (ERG) was used to evaluate the retina functional variation, and the apoptotic rate of photoreceptor cells was determined by TUNEL staining. HE staining was performed. The number of photoreceptor cells of the outer nuclear layer was calculated. Furthermore, effect of WYHR on Rhodopsin and basic fibroblast growth factor (bFGF) expression was examined using immunochemical assay.

RESULTSCompared with the model group, a- and b-wave latency and amplitude, as well as the bFGF expression sharply increased in the Max-ERG of the Chinese medicine group (P < 0.05, P < 0.01) at 18 postnatal days. At 28 and 48 postnatal days, a- and b-wave latency and amplitude sharply increased, photoreceptor cell layer numbers of the outer nuclear layer obviously increased, the apoptosis rate of retinal photoreceptor cells obviously decreased, expressions of Rhodopsin and bFGF in the Chinese medicine group significantly increased (P < 0.05, P < 0.01).

CONCLUSIONWYHR could effectively inhibit the apoptosis of photoreceptor cells in RDS mice, which might be attributed to up-regulating bFGF expression.

Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Photoreceptor Cells, Vertebrate ; cytology ; drug effects ; Retinal Degeneration ; metabolism ; pathology

Adult ; Antiviral Agents ; pharmacology ; therapeutic use ; DNA, Viral ; Female ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate the clinical application value of throat swab nested PCR for detecting active congenital human cytomegalovirus (HCMV) infection in neonates.

METHODSThe throat swabs and umbilical cord blood specimens from 51 neonates were collected for nested PCR assay for HCMV glycoprotein B (gB) gene. Moreover, 18 of them were subjected to a pp65 antigen test.

RESULTSThe sensitivity and specificity of throat swab nested PCR for HCMV gB gene were 67% and 75%, respectively, and the positive and negative predictive values were 57% and 82%, respectively.

CONCLUSIONSThroat swab nested PCR assay for HCMV gB gene is non-invasive, rapid, and highly sensitive for HCMV detection and holds promise as an excellent screening technology for detecting active congenital HCMV infection in neonates.

Cytomegalovirus Infections ; congenital ; diagnosis ; Fetal Blood ; virology ; Humans ; Infant, Newborn ; Phosphoproteins ; blood ; Polymerase Chain Reaction ; methods ; Viral Envelope Proteins ; genetics ; Viral Matrix Proteins ; blood

OBJECTIVETo explore the relationship between CD8+ T-cell CD28 molecular expression in peripheral blood and TCM type in patients with chronic aplastic anemia (CAA).

METHODSUsing flow cytometry to detect the CD28 expression in 45 in-patients or out-patients and 24 healthy subjects for control. And the relation with TCM type was analyzed from the immunological aspect.

RESULTS(1) The levels of CD8, CD28, CD8+ CD28+ expression and CD8+ CD28+/CD8+ CD28- were all higher in the CAA patients than those in the healthy subjects (P < 0.05 or P < 0.01). (2) The levels of CD28, CD8+ CD28+ expression and CD8+ CD28+/CD8+ CD28- were all higher in the CAA patients of Shen-Yin deficiency type than those in the CAA patients of Shen-Yang deficiency type (P < 0.05 or P < 0.01).

CONCLUSION(1) The abnormal high expression of peripheral blood co-stimulatory molecules CD28 suggested CD28 disorder may play an important role in immuno-pathogenesis of CAA. (2) The levels of peripheral CD28, CD8+ CD28+ expression and CD8+ CD28+/CD8+ CD28- can be taken as an objective indexes for TCM typing of CAA, which was disordered more severe in patients of Shen-Yin deficiency type than in those of Shen-Yang deficiency type.

Adult ; Anemia, Aplastic ; classification ; diagnosis ; immunology ; CD28 Antigens ; biosynthesis ; CD8-Positive T-Lymphocytes ; immunology ; Chronic Disease ; Diagnosis, Differential ; Humans ; Kidney Diseases ; immunology ; Male ; Medicine, Chinese Traditional ; Yang Deficiency ; immunology ; Yin Deficiency ; immunology

Objective: To explore age, gender, and regional differences in physical activity among children and adolescents in China, and to provide a scientific reference for enhancing physical activity promotion. Methods: A total of 4 269 children and adolescents aged 7 to 18 years were selected from six administrative regions of China (East China, Northwest China, North China, Central China, Southwest China and South China) using a stratified random cluster sampling method from September to December 2018. A questionnaire was administered to evaluate the physical activity level of Chinese children and adolescents aged 7 to 18. Results: The overall detection rate of MVPA insufficiency in children and adolescents in China was 53.8%, of which the detection rate of MVPA insufficiency was 50.8% among boys and 57.1% among girls. Gender differences were statistically significant ( χ 2= 17.10 , P <0.05). Among the different age groups, the lowest detection rate of MVPA among 10-12 year olds was 43.6%, whereas the highest rate among 16-18 year olds was 63.0%, with significant differences between gender ( χ 2=4.33, 30.79, P <0.05). The P 50 values of total physical activity(TPA), light intensity physical activity(LPA), moderate intensity physical activity(MPA), vigorous intensity physical activity(VPA), moderate to vigorous physical activity(MVPA) were 92.9,24.3,41.4,7.1 and 55.7 min/d , respectively. The P 50 values of physical exercise, housework activities, entertainment activities and transportation activities were 34.3 , 2.1, 2.3 and 30.0 min/d, respectively, and the difference in age groups was statistically significant( H =95.03, 74.99, 300.26 , 64.16, P <0.05). There was a statistically significant difference in the detection rate of insufficient MVPA among children and adolescents in different regions ( χ 2=83.91, P <0.05). The lowest rate was 44.0% in North China, and the highest was 65.9% in East China. Conclusion The detection rate of MVPA insufficiency among Chinese children and adolescents firstly decreased and then increased with age. Boys participated in higher levels of physical activity than girls.

OBJECTIVETo assess the larvicidal effects of recombinant Escherichia coli expressing scorpion neurotoxin AaIT or Bacillus thuringiensis subsp israelensis (B.t.i) toxin Cyt2Ba against the second instar larvae of Culex pipiensquinquefasciatus and Aedes albopictus and compare different formulations for their larvicidal effects.

METHODSThe AaIT- or Cyt2Ba-coding sequences were cloned into pET28a(+) and the recombinant plasmids were transformed into E. coli BL21(DE3). After induction with IPTG, the recombinant proteins expressed by the recombinant E. coli were detected and identified by SDS-PAGE and Western blotting, respectively. The larvicidal activity of the bacterial suspension was tested at different concentrations against mosquitoes. The effective engineered bacteria were prepared into dry powder with different formulations, and their larvicidal activity was tested.

RESULTSAaIT and Cyt2Ba proteins were successfully expressed in E. coli. The recombinant AaIT protein showed no virulence to the mosquito larvae. The suspension of the recombinant E. coli expressing Cyt2Ba protein exhibited a stronger killing effect on Aedes albopictus larvae than on Culex pipiens quinquefasciatus larvae at 48 h (P<0.001) with LCof 3.00×10cells/mL and 1.25×10cells/mL, respectively. The dry powder of the engineered bacteria formulated with yeast extract, wheat flour or white pepper powder at the mass ratio of 1:1 showed the strongest killing effect on mosquito larvae (P=0.044), and the formulation with white pepper powder produced a stronger killing effect than formulations with yeast extract or wheat flour (P=0.002).

CONCLUSIONThe B.t.i Cyt2Ba protein expressed in E. coli BL21(DE3) shows a good larvicidal activity against mosquitoes, and appropriate formulations of the engineered bacteria can enhance its efficiency in mosquito control.

The spike (S) glycoprotein of HCoV-NL63 is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. Four fragments coding spike proteins (S1, S2, RL and RS) from HCoV-NL63 were amplified and cloned into the expression vector derived from vaccinia virus (Tiantan strain), and recombinant vaccinia viruses expressing four segments of spike proteins were generated (vJSC1175-S1; vJSC1175-S2; vJSC1175-RL; vJSC1175-RS), respectively. Their expression location in cell and level were characterized using indirect immune fluorescence assay (IFA) and Western-Blot, respectively. The expressions of four segments of spike proteins in recombinant vaccinia viruses were showed at appropriate level and with posttranslational modification (glycosylation), and S1, RL and RS were mainly distributed in the cell membrane, while the S2 was mainly distributed in the cytoplasm. Our results provide a basis for further exploring diagnostic role and vaccine development of different spike segments from HCoV-NL63.
Base Sequence ; Blotting, Western ; Coronavirus NL63, Human ; chemistry ; Fluorescent Antibody Technique, Indirect ; Humans ; Membrane Glycoproteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Plasmids ; Recombinant Proteins ; biosynthesis ; Spike Glycoprotein, Coronavirus ; Vaccinia virus ; genetics ; Viral Envelope Proteins ; biosynthesis ; genetics

OBJECTIVETo explore the techniques of esophagogastrostomy or esophagojejunostomy in the mediastinum through the abdomen and hiatus after extended proximal gastrectomy or total gastrectomy.

METHODSFrom May 2010 to January 2012, 15 patients with esophagogastric junction carcinoma underwent open transhiatal extended gastrostomy or total gastrectomy. After full mobilization, the anvil was reversely introduced into the esophagus and the esophagus was transected with curved stapler. The rod of the anvil was then pulled out with a stitch to complete esophagogastrostomy after proximal gastrectomy(n=9) or esophagojejunostomy after total gastrectomy(n=6).

RESULTSThe anastomosis was successfully performed in all the patients. The mean operation time was(185.5±13.1) min. The mean operation time for anastomosis was(42.0±8.6) min. The mean estimated blood loss was (106.7±34.9) ml. The proximal resection margin was(4.4±1.2) cm. All the margins were negative for residual cancer. There was no postoperative death or fistula. During the follow up, there was one case of anastomotic stenosis which was successfully managed by endoscopic balloon dilatation.

CONCLUSIONSEsophagogastrostomy or esophagojejunostomy can be safely performed with double stapling technique including reverse anvil introduction and curved stapling transection of the esophagus. It is an ideal technique for anastomosis after extended gastrectomy for esophagogastric junction carcinoma.

Aged ; Anastomosis, Surgical ; methods ; Esophagogastric Junction ; Esophagus ; surgery ; Female ; Gastrectomy ; Humans ; Jejunum ; surgery ; Male ; Middle Aged ; Stomach Neoplasms ; surgery