It was found that pre-X region was present in five clones of hepatitis B virus (HBV) genome from 2 patients with chronic HBV infection. The open reading frame (ORF) consisted of 168 base pair (bp), and could be transslated with the X gene in frame. To investigate the activity of pre-X gene promoter, promoter DNA sequence was amplified from HBV-DNA by polymerase chain reaction (PCR), The amplified product was cloned into pCAT3 vector, The HepG2 cells were transfected by pCAT3-Pre-X-p. The CAT activity was detected by an enzyme-linked immunosorbent assay (ELISA) kit. It was found that pCAT3-Pre-X-p had higher activity of CAT, which was 3.5 fold over that of pCAT3-promoter. This result implicated that pCAT3-Pre-X-p possessed promoter activity.

To investigate the promoter activity of pre-pre-S gene, which was found in five clones of hepatitis B virus (HBV) genome from 2 patients with chronic HBV infection, promoter DNA sequence was amplified from HBV-DNA by polymerase chain reaction (PCR).The amplified product was cloned into pCAT3 reporter vector. The HepG2 cells were transfected by pCAT3-pre-pre-S-p. The CAT activity was detected by an enzyme-linked immunosorbent assay (ELISA) kit. It was found that pCAT3-pre-pre-S-p had higher activity of CAT which was 10 fold over that of pCAT3-basic. This result implicated that pCAT3-pre-pre-S-p had promoter activity.

Objective To screen and clone the target genes transactivated by nonstructural protein 5A (NS5A) of hepatitis C virus (HCV). Methods The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-NS5A and pcDNA3.1(-) empty vector, respectively. Suppression subtractive hybridization (SSH) method was employed to analyze the differentially expressed DNA sequence between the two groups. The obtained sequences were searched for homologous DNA sequence from GenBank. The new gene with no homology with known genes in this database was confirmed, and electric polymerase chain reaction was conducted for cloning the full-length DNA of the new gene and in conjunction with Kozak rule and the terminus of polyadenyl signal sequence. The reverse transcription PCR (RT-PCR) was used to amplify the new gene from mRNA of HepG2 cell as the template. The coding sequence of the new gene was deduced according to the nucleotide sequence. Results A new gene with unknown function was named as NS5ATP3. The nucleotide sequence of the NS5ATP3 gene and its corresponding amino acid have been determined, which contained 1 572nt and 524aa. The sequence of the NS5ATP3 gene was deposited into GenBank, with the accession number AF529364. Conclusions NS5ATP3 gene transactivated by HCV NS5A protein was cloned and identified successfully by combining molecular biological technology and bioinformatics technique. These results will pave the way for the study of the molecular mechanism of the transactivating effects of HCV NA5A protein and the development of new therapy for chronic hepatitis C.

Objective To investigate the activating effect of HBV X protein on transcription of calgizzarin S100A11 gene promoter. Methods The sequence of calgizzarin S100A11 gene promoter was identified in GenBank by bioinformatics and amplified from HepG2 genome by polymerase chain reaction (PCR). The amplified product was cloned into pCAT3 reporter vector. The HepG2 cells were transfected by pCAT3-S100-p, and then co-tranfected by pCAT3-S100-p and pcDNA3.1(-)-X. The choloraphenical acetyltransferase(CAT) activity was detected by enzyme-linked immunosorbent assay(ELISA) kit. Results It was shown that pCAT3-S100-p could regulate the expression of CAT in HepG2 cells. The expression of CAT in co-transfection of pCAT3-S100-p and pcDNA3.1(-)-X was 2.1 fold higher than pCAT3-S100-p plasmid. Conclusion HBV-X protein can trans-activate the expression of S100A11 protein, and it further proved our previous results by SSH and biochips.

Objective:To longitudinally analyze the unit costs and technical efficiency of human immunodeficiency virus(HIV) voluntary counseling and testing in China.Methods:Unit costs were calcula-ted by the province and period using longitudinal data from 7 provinces covered by Global Fund China AIDS Program Round 3,and then technical efficiency and Malmquist indices were measured with an approach to data envelopment analysis.Results:The unit costs for HIV voluntary counseling and testing changed dramatically over a 3+-year period,decreasing from $165.97(mean) to $53.41,with an accumulative unit cost of $67.19,and its technical efficiency was averaging between 0.44 and 0.63.Conclusion:The time series of unit costs for HIV voluntary counseling and testing formed a U-shape curve with an inflection point before which unit costs dramatically dropped and another inflection point beyond which unit costs went up.These findings can inform program managers of the changing unit costs when extending or expanding HIV prevention efforts.

OsRacD is a critical molecular switch involved in photoperiod fertility conversion of photoperiod sensitive genic male sterile rice.Taking OsRacD as bait,a new rice myosin heavy chain partial cDNA was isolated from the rice panicle by yeast two-hybrid,and designated OsMY1.The interaction between OsMY1 and OsRacD had been testified by yeast two-hybrid and pull down assay,it showed that OsMY1 could bind with OsRacD,suggested that OsMY1 maybe the target of OsRacD.The important evidence for further study on the functional relationship between OsMY1 and OsRacD in vivo are also provided.

Cell immobilization is a new biotechnology The definition, classification, and carrier selection of cell immobilization are presented in details The technique is efficiently applied to treating strength organic wastewater, nutrient and heavy metals removal of wastewater, as well as hardly biodegradated wastewater It has a widely applied prospect in wastewater treatment

Objective To observe the therapeutic effect of antiangiogenesis on C6 glioma and to study the practical value of PWI in evaluating the early response of the tumor to the treatment.Methods C6 glioma cells were stereotactically implanted into SD rats intracranially.At the 15 day after tumor implantation,the recombinant human endostatin was administered or stereotactic radiosurgery was performed.Endostatin was injected subcutaneously at the dose of 5 mg?kg -1 ?d -1 ,10 mg?kg -1 ?d -1 ,and 20 mg?kg -1 ?d -1 for 7 days.Stereotactic radiosurgery was given once with the central radiation dose of 25Gy.MR PWI was performed to measure relative cerebral blood volume (rCBV) of the tumor and the normal brain before and 48 hours after treatment.Conventional MRI was performed to calculate tumor volume.Results The tumor volume was (429.0?36.7)mm 3,(305.7?32.8)mm 3,and (277.0?20.6)mm 3 in each endostatin treated group,and(390.0?33.8)mm 3 in SRS treated group,which were lower than that in control group(566.7?135.0)mm 3 (P=0.031).The rCBV in endostatin group was 1.57?0.12,1.30?0.12,and 1.24?0.08,respectively according to three different doses,which were lower than those before endostatin therapy with statistical significance (P

Objective To explore the dynamic variation rule of drainage fluid parathyroid hormone ( dPTH) , serum parathyroid hormone ( PTH) and serum calcium after thyroidectomy .According to the variation rule, the survival , function and prognosis of the parathyroid which retained at the original place can be predicted . Methods From Apr.2012 to Aug.2012, 90 patients who underwent thyroidectomy in Thyroid Surgery Center of the First Affiliated Hospital of Kunming Medical University were chosen as the research object .All of the objects'operations were performed by the same surgeon team and they were divided into four groups according to different surgical methods ( group A:bilateral thyroidectomy group , group B:bilateral thyroidectomy and central neck dis-section group , group C: bilateral thyroidectomy and functional neck dissection group , and group D: unilateral thyroidectomy group ) .Parathyroid retention situation during operation and the occurrence of hypocalcaemia after operation were recorded .Blood samples were taken between 7am and 8am in the 4 consecutive days after opera-tion to detect serum calcium and serum parathyroid hormone .All of the 90 patients had the drainage tube and their drainage fluid were taken for testing dPTH .The levels of serum calcium , PTH and dPTH were analyzed by statistical analysis of repetitive measure analysis of variance ( ANOVA) .Results The mean postoperative maxi-mum serum calcium was in group D and the minimum was in group C .There was no obvious difference in terms of serum calcium between group A and group B .However , the serum calcium showed an upward trend in each group.The mean postoperative maximum PTH was in group D and the difference has statistical significance com -pared to the other 3 surgical methods.The minimum was in group B and group C .However, it showed an upward trend with time in each group .The difference of dPTH in each group had no statistical significance and it showed a downward trend in all the 4 groups.Low serum calcium and hypocalcaemia occurred to 22 cases and 13 cases respectively after operation .The low serum calcium cases in each group were 12, 3, 4 and 2 respectively and hy-pocalcaemia cases in each group were 4, 3, 1 and 1 respectively.Conclusions By monitoring dPTH, PTH and serum calcium after thyroidectomy , the survival and function of parathyroid retained at the original place can be e-valuated comprehensively .Furthermore, it also helps to estimate prognosis .dPTH at a high level after operation is a direct evidence that parathyroid retained at the original place survives .Low PTH and high dPTH after thyroid-ectomy illustrates the operation just affects the way that PTH secreted into blood and the parathyroid retained at the original place can still secrete large amount of PTH .PTH will return to normal gradually with reconstruction and healing of microcirculation around parathyroid .Persistent low serum PTH after operation , low dPTH after 24 hours and the early advent of hypocalcaemia suggest the parathyroid retained at the original place was injured seri -ously and its blood supply was damaged obviously and more than one parathyroid were affected .Secretion function of parathyroid will remarkably decrease .

Abstract: Objective　To analyze the treatment outcomes of elderly patients with pulmonary tuberculosis in Chongqing, so as to provide reference for the prevention and control strategies of the epidemic of elderly pulmonary tuberculosis. Methods　The data of tuberculosis cases aged ≥65 years in Chongqing from 2015 to 2020 were collected from the National Health Insurance Information Project Disease Prevention and Control Information system. Descriptive statistical methods were used to analyze the data. Results The registration rates of elderly active pulmonary tuberculosis patients and etiological positive patients were 110.95/10-5and 32.25/10-5 in 2015 and 84.06/10-5 and 57.29/10-5 in 2020. The annual decline rate of active tuberculosis registration was 5.40%, and the annual increase rate of pathogenic-positive tuberculosis registration was 12.18%. The registration rates of active tuberculosis patients and etiological positive patients in the whole population were 70.75/10-5 and 17.63/10-5 in 2015 and 50.34/10-5 and 29.14/10-5 in 2020. The annual decline rate of active tuberculosis registration was 6.58%, and the annual increase rate of pathogenic-positive tuberculosis registration was 10.57%. From 2015 to 2020, a total of 25 931 cases of elderly pulmonary tuberculosis were registered, of which 21 374 (82.43%) cases were successfully treated and 4 010 (15.80%) cases had unfavorable outcomes. The proportion of cured and death patients showed an increasing trend year by year (χ2trend=313.853, 100.502, P<0.01). From 2015 to 2020, the average annual successful treatment rate of elderly pulmonary tuberculosis in the whole city was 82.43%, with the lowest rate in southeast Chongqing (74.23%), followed by urban areas (81.99%). The success rate of elderly pulmonary tuberculosis treatment in the whole city, west Chongqing, northeast Chongqing and southeast Chongqing showed a downward trend year by year (χ2trend=230.199, 35.278, 108.076, 112.130, all P<0.01), with annual decline rates of 2.77%, 2.26%, 3.0% and 4.12%, respectively. Among the registered elderly patients, female, 65-<75 years old, Han nationality, newly diagnosed, no complications, and negative for etiology (χ2=15.234, 255.910, 146.842, 179.998, 25.575, 131.170, P<0.01) had higher success treatment rates. Conclusions The prevalence of pulmonary tuberculosis in the elderly population in Chongqing City is declining, but the positive registration rate of etiology is increasing annually, and the success rate of treatment is decreasing. Therefore, it is necessary to strengthen the systematic management, publicity and education of elderly patients (especially those in southeast Chongqing, male, positive patients and severe patients) to effectively control the epidemic of tuberculosis in the elderly.