Objective:To synthesize Gd labeled probe targeting transporter protein(TSPO) 2-(8-amino-2-(4-chlorophenyl)imidazo[1, 2-a]pyridine-3-yl)- N, N-dipropylacetamide (CB86)-diethylene triamine pentaacetic acid (DTPA), and investigate its MRI effect in rheumatoid arthritis (RA) model. Methods:CB86-DTPA was prepared by coupling a bifunctional chelating agent, and then chelated with Gd to obtain MRI targeted contrast agent CB86-DTPA-Gd. The cytotoxicity, MR relaxation rate and in vitro stability of CB86-DTPA-Gd were determined. RA model was established with Freund′s adjuvant and the biodistribution study and MRI was performed. The RA lesion and its surrounding normal tissue were used as regions of interest (ROI) to calculate the signal to noise ratio (SNR). Independent-sample t test was used to analyze the data. Results:CB86-DTPA-Gd had excellent biosafety and a good MR relaxation rate ( r1=11.05 mmol·L -1·s -1). The survival rate of RAW264.7 cells and 4T1 cells was still more than 90% at the maximum concentration (20 μmol/L) of Gd 3+. CB86-DTPA-Gd also exhibited good stability in human serum and phosphate buffer saline solution (PBS; pH=7.4). The in vivo biodistribution showed that CB86-DTPA-Gd had better inflammatory targeting efficiency, and the uptake of Gd in the inflamed site of the ankle joint was still (2.33±0.29) percent dose rate per gram of tissue (%ID/g) at 120 min after injection. MicroMRI showed that the inflammation of the right ankle joint displayed significant enhancement after the injection of CB86-DTPA-Gd and Gd-DTPA. The SNR of CB86-DTPA-Gd group was up to 23.21±1.44, and the maximum intensification time was 90 min after injection, and can be significantly inhibited by CB86-DTPA at all time points ( t values: 6.083-12.451, all P<0.05), while the Gd-DTPA group had a strengthening time of 30 min after injection with the SNR of 16.12±1.24. Conclusion:CB86-DTPA-Gd shows good macrophage targeting and good uptake in arthritic reaction sites, and is expected to be a novel MRI molecular probe for peripheral inflammation imaging.

Objective To study whether the organs besides digestive system of musca domestica Ⅲ instar larvae have the capability of produceing musca β-glucosidase.Methods Tissues of malpighian tubules,trachea,epiploon and body wall of musca domestica Ⅲ instar lar-vae were dissected under anatomic microscope,and the expression of β-glucosidase gene in these dissected tissues were detected by reverse transcription PCR.And the tissue localization of β-glucosidase mRNA was further identified by in situ hybridization.Moreover,anti-cellulase was used to determinate the tissue distribution with immunohistochemical staining.The relative mRNA expression levels of musca domesticaβ-glucosidase gene in these organs were tested by real-time quantitative PCR.Results The reverse transcription PCR showed that the ampli-fication products of β-glucosidase gene were observed in tissues of malpighian tubules,trachea and body wall.β-glucosidase mRNA was shown in the epithelium cells of malpighian tubules,trachea and body wall by in situ hybridization,and it was almost the same in the results of im-munohistochemical staining.The real-time quantitative PCR showed that the relative expression quantity of β-glucosidase gene in malpighian tubules and body wall were higher than that in foregut,while it was lower in itrachea than that in foregut.And it was of statistical difference in gene expression level of β-glucosidase among these organs (P <0.05).Conclusion Malpighian tubules,trachea and body wall of musca domestica Ⅲ instar larvae have the function of secreting β-glucosidase.Combining with the characteristics of secreting β-glucosidase in most organs of digestive system,it may provide a new biological method for the prevention and treatment of human diseases transmitted by musca domestica with the use of taget gene β-glucosidase.

Objective To investigate the prevalence and risk factors of chronic kidney disease (CKD) in Chengdu urban population and the prevalence of CKD in risk population.Methods Questionnaire (anamnesis,smoking,drink) of risk factors of CKD and somatoscopy (blood pressure,body height and body weight) were caried out in railman of Chengdu urban.Their blood and urine indicators (blood sugar,blood lipid,blood uric acid,blood creatinine,uromicroprotein/creatinine ratio,routine urine examination,etc) were measured.The prevalence and risk factors of CKD in Chengdu urban population and the prevalence of CKD in risk population were elucidated.Results Eligible data of 5326 subjects were enrolled in the study.After the adjustment of age and gender component,the prevalence of albuminuria was 11.54％,reduced eGFR was 5.54％,hematuria was 3.87％,and CKD was 18.32％; the recognition was 1.93％.In addition,the prevalence of albuminuria was respectively 23.79％,28.00％,14.08％; prevalence of reduced eGFR was respectively 4.76％,4.53％,3.26％; prevalence of hematuria was respectively 2.94％,3.20％,2.37％ in 3098 people with hypertension,diabetes or hyperlipaemia.Independent risk factors of albuminuria were female,hypertension,diabetes,hyperlipemia and high BMI.Independent risk factors of reduced eGFR were female,age,hyperuricemia and hypertension.Drink was negatively correlated with reduced eGFR.Independent risk factors of hematuria were female and age.Conclusions The prevalence of CKD is quite high and the recognition rate is low in the Chengdu urban populaton.Risk factors of CKD are age,female,diabetes,hypertension,hyperlipemia,hyperuricemia and high BMI.Control of the development of metabolic disease can reduce the CKD.

BACKGROUND: Polymorphism in promoter region can change the expression of genes, which may be associated with susceptivity of diseases. Gene polymorphism of interleukin-6 (IL-6) promoter is associated with nationality and many diseases. Different nationalities often display different characteristics of gene polymorphism.OBJECTIVE: To observe the distribution of polymorphism at sites -597 and -572 in IL-6 promoter region in Tibet Tibetan population and to provide the theoretical data for Tibetan population genetics and background of immunity.DESIGN: Randomized investigation.SETTING: Institute of Anthropology, Jinzhou Medical College.PARTICIPANTS: Totally 108 healthy Tibetan teenagers were selected from Lasa and Naqu region in Tibet autonomous region from October 2003 to July 2004, including 60 males and 48 females, aged from 14-21 years. Inclusive criteria:The parents of the volunteers were healthy Tibetans after body examination. The volunteers knew the fact, agreed to participate into the trail and signed the informed consent.METHODS: 5 mL peripheral vein blood was collected from 108 Tibetan teenagers. DNA from human leucocytes was extracted by salt fractionation. IL-6 promoter including -597 and -572 fragments was amplified by polymerase chain reaction (PCR). Representative fragments were cloned then sequenced after restriction fragment length polymorphism (RFLP).MAIN OUTCOME MEASURES: Distribution of polymorphism in Tibet Tibetan population; Results after comparison with those of other nationalities including Han population.RESULTS: Data of 108 Tibetan teenagers were involved in the result analysis. ①Distribution of polymorphism on -572C/G site of IL-6 promoters in population of either sex: There were no GA and AA genotypes at site -597, but only GG genotype appeared. There were CC, CG and GG genotypes at site -572, and the frequencies were 0.63, 0.35 and 0.02 in order. Distribution of genotype with representativeness met the Hardy-Weinberg equilibrium. Allele frequencies were 0.81 and 0.19, respectively. There was no significant difference of either sex in genotype and allele frequency (P ＞ 0.05). ②Distribution of polymorphism on -597 G/A and -572 C/G in different nationalities: GG, GA, AA genotypes appeared on -597 site in England and France, and G and A allele frequencies were 0.60 and 0.40,respectively. It was significantly different from that of Tibetan in Tibet. Furthermore, Japanese had no polymorphism,which was similar to that of Hans in China (P ＞ 0.05). ③Genotype of different straps and results of DNA sequencing:Only GG genotype was found on -597 site (without the restriction site, one fragment after restriction, PCR amplification products), no GA and AA genotypes. CC, CG and GG genotypes appeared at site -572, and frequencies were 0.64,0.35 and 0.01, respectively. Distribution of genotype with representativeness met the Hardy-Weinberg equilibrium (P ＞0.05). Allele frequencies were 0.81 and 0.19, respectively. There was no significant difference of either sex in genotype and allele frequency. Distribution of gene frequency and allele frequency in IL-6 were similar between Tibetan and that of Hans, but it was significantly different from that of population in England, France and America.CONCLUSION: There are nationality differences of IL-6 gene polymorphism at sites -597 and -572. No polymorphism is found at site -597 in Tibetan. Race differences are seen at site -572, having CC, CG and GG genotypes and G allele is rate. Compared with white population, there is significant difference in genotype and allele frequency at site -572. Their characteristics are close to Hah population and Japanese, which may be associated with genetic gene of persons living in plateau.

Objective:To study the antimalarial mechanism of benflumetol (B). Methods: Flow cytometry (FCM) was used to analyze the effects of B and chloroquine (CQ) on DNA content of Plasmodium berghei and pH value of the lysosome of malarial parasites. Results: DNA content of the plasmodia not treated with any drugs was not changed in 24 hours,while benflumetol could decrease the DNA content: the DNA content began to decrease 2 h after the drug administration and reached the minimum by 16 h, but somewhat increased at 24 h after administration. The pH in the lysosome increased 1 h and restored premedication level 4 h after benflumetol administration. Chloroquine had the same effects on DNA and lysosome pH of malarial parasites.Conclusions: The antimalarial mechanism of benflumetol is directly related to its effect to inhibit the synthesis of DNA.

Objective: To learn the concentration of target genes of severe acute respiratory syndrome coronavirus 2 ( SARS-CoV-2 ) and the influencing factors in the confirmed cases with coronavirus disease 2019 ( COVID-19 ) in Ningbo. Methods: Demographic information and clinical diagnosis of COVID-19 cases reported in Ningbo from January 21 to February 20, 2020 were collected through China Disease Control and Prevention Information System. The Ct values of ORF1ab and N of the cases were collected through Ningbo Center for Disease Control and Prevention and designated hospitals, and analyzed in terms of gender, age, severity, clinical classification, source of case, sampling time and specimen type. Results: There were 157 confirmed COVID-19 cases, including 56 males ( 35.67% ) and 101 females ( 64.33% ). Sixty-seven cases ( 42.68% ) aged between 40 and 60 years old. The Ct values of ORF1ab and N in females were 29.96±5.28 and 30.38±4.90, which were higher than 27.56±4.94 and 28.03±4.88 in males ( P<0.05 ). The Ct values of ORF1ab and N were the lowest when sampled at 1-7 days after onset ( P<0.05 ), which were 27.84±4.80 and 28.35±4.65. The Ct values of ORF1ab ( rs=0.288, P=0.001 ) and N ( rs=0.296, P=0.001 ) were positively correlated with sampling time. The Ct values of ORF1ab in pharyngeal swab and sputum were 29.19±4.85 and 28.74±6.40, the Ct values of N in pharyngeal swab and sputum were 29.61±4.60 and 29.22±6.10, both without significant differences between different samples ( P>0.05 ). Conclusions Sampling time has a great influence on the Ct values of ORF1ab and N of SARS-CoV-2.

The cDNA coding spinach glycolate oxidase (GO) was amplified by RT-PCR using the total RNA of spinach leaves as the template, and was cloned into cloning vector pMD18-T. After the DNA sequence was determined, the go gene was subcloned into E. coli expression vector pBV220, pET-22b(+), pTIG-Trx and pThioHisC. SDS-PAGE analysis revealed that the recombinant E. coli BL21 (DE3) (pTIG-Trx-GO) and E. coli BL21 (DE3) (pET-22b(+)-GO) expressed the predicted 38 kD glycolate oxidase, and the enzyme activity was also detected.
Alcohol Oxidoreductases ; genetics ; isolation & purification ; metabolism ; Cloning, Molecular ; Escherichia coli ; Gene Expression ; drug effects ; Isopropyl Thiogalactoside ; pharmacology ; Spinacia oleracea ; enzymology ; genetics

Weightlessness environment can lead to the muscle atrophy and body fluid distribution upward,which can cause the bone calcium metabolism disorder and always accompanied by the loss of bone microstructure and increased rate of bone fracture. Under microgravity,the astronauts are much easier to decrease the Ca2+ ion in bone, which can cause serious osteoporosis. However the bone lost is not equilibrium, it is especially serious in the mechanism loading bone and the recovery process is more difficult. These are very different from the osteoporosis in older people and postmenopausal osteoporosis. It is necessary to find an optimal method to due with it. In traditional Chinese medicine theory,the kidney stores "Jing" and dominates the bone, thus a lot of bone related diseases can be treated through the kidney. A lot of clinical practices have also proved that the Chinese herbs used under the guidance of basic Chinese medicine theory are always good at the treatment of common osteoporosis. In simulated weightlessness experiment, people found that the kidney nourishment drugs do can prevent the decrease of BMD. So in this article we want to review the causes of weightlessness and the potentials applications of tradition Chinese medicine in the treatment of weightlessness osteoporosis.
Humans ; Mechanical Phenomena ; Medicine, Chinese Traditional ; methods ; Osteoporosis ; etiology ; prevention & control ; therapy ; Weightlessness ; adverse effects

In order to establish a method by which the recombinant suicide plasmids integrated on the chromosome could be recircled, A simple method of transconjugative cloning was established with the helper plasmids pMT999 or pRK2013 and fusion strains of Shigella flexneri which were obtained by screening with in vivo expression technology. And the cloning efficiency with this method is very high.