To construct a subtractive cDNA library of genes transactivated by NS5A protein of hepatitis C virus with suppression subtractive hybridization technique, the mRNA was isolated from HepG2 cells transfected with pcDNA3 1(-) NS5A and pcDNA3 1(-) empty vector, respectively, then the cDNA was synthesized. After restriction enzyme RsaI digestion, small sized cDNAs were obtained. Then the tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. After the tester cDNA was hybridized with the driver cDNA twice and underwent two times of nested PCR, it was then subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. The amplified library contained 121 positive clones. Colony PCR showed that 115 clones contained 200- 1 000 bp inserts. Sequence analysis was performed in 90 clones, and the full length sequences were obtained with bioinformatics method. Altogether 46 kinds of coding sequences were acquired, which consisted of 31 kinds of known and 15 kinds of unknown ones. The obtained sequences might be target genes transactivated by NS5A protein of HCV, among which some genes coding proteins involved in cell cycle regulation, cell apoptosis, signal transduction pathway and tumour development. The results indicated that the subtractive library of genes transactivated by NS5A protein of HCV was constructed successfully, which brought some new clues for studying the biological functions and pathogenesis of the viral proteins

To evaluate the protective effect of MgSO_4 during cerebral ischemia reperfusion,twentyfour SD rats were divided randomly into control group (n=7),ischemic reperfusion group (n=9)and MgSO_4 treatment group (n=8)with Pnlsinelli and Brierley's animal ischemic reperfusion model. Compared with the control and ischemic reperfusion group,cerebral tissue contents of water and malondialdehyde reduced markedly (P

Objective To study the epidemiological and clinical features of human parainfluenza virus (HPIV) infection in children in Suzhou,and to provide the evidence-based foundation for early warning,diagnosis and treatment of respiratory infection in children.Methods The sputum specimens and medical history were obtained from children with acute respiratory tract infection hospitalized at the Childen's Hospital Affiliated to Soochow University from January 2006 to December 2015.Seven kinds of common respiratory viruses including respiratory syncytial virus,influenza virus A,influenza virus B,HPIV Ⅰ,HPIV Ⅱ,HPⅣV Ⅲ and adenovirus were detected by using the direct im-munofluorescence.Mycoplasma pneumoniae(MP),chlamydia pneumoniae,human bocavirns (hBoV) were detected by using fluorescence quantitative PCR.Rhinovirus and human metapneumovirus were detected by using reverse transcription-PCR.Sputum was cultured for bacteria.Results In 21 769 cases,the detection rate of HPIV positive was 3.21％ (829 cases),among which,HPIV Ⅰ,HPIV Ⅱ,HPIV Ⅲ were respectively detected in 113 cases (0.52％),16 cases (0.07 ％) and 700 cases (3.21％),respectively.There were 378 cases of simple infection and 428 cases of mixed infection,and the mixed infection was very common in Streptococcus pneumoniae,Haemophilus influenza,MP and hBoV.There was a difference in HPIV infection among genders,and the detection rate of the boys was higher than that of girls[4.14％ (563/13 591 cases) vs.3.25％ (266/8 178 cases),x2 =11.036,P =0.001].In the 28 d-1 year old and ＞ 1-3 year old group,the detection rate of HPIV was higher[4.71％ (494/10 476 cases) and 4.21％ (244/5 793 cases),respectively].In spring and summer,there was a higher detection rate of HPIV infection.The clinical manifestations with simple infection of HPIV Ⅰ and HPIV Ⅲ were cough,fever and wheezing.The rate of fever and shortness of breath in those of HPIV Ⅰ was 71.74％ (33/46 cases),10.87％ (5/46 cases),and that in HPIV Ⅲ was 40.12％ (134/334 cases),2.10％ (7/334 cases),HPIV Ⅰ infection was more likely to cause fever and shortness of breath than those of HPIV Ⅲ,there were significant differences (x2 =16.410,P ＜ 0.001;x2 =10.177,P =0.001).Pneumonia had the highest detection rate of viral infection.Conclusions HPIV Ⅲ is the leading pathogen among the types of HPIV in the hospitalized children in Suzhou area.Among the subtypes of HPIV,the peak of HPIV infection occurs in spring and summer.The children less than 3 years old are the most susceptible to parainfluenza virus,and the HPIV detection rate is gradually declines with age.

Objective To analyze the clinical features of infants infected by respiratory syncytial virus (RSV) or human rhinovirus (HRV) in lower respiratory tract in Suzhou area based on the month age and the month of the year.Methods From January 2013 to December 2015,2 206 nasopharyngeal aspirates specimens were collected from the infants with lower respiratory tract infection.Direct immunofluorescence assay was performed to test RSV.Reverse transcription-polymerase chain reaction(RT-PCR) method was used to test HRV.The medical history was collected and pulmonary function tests were performed in some infants who were infected with RSV and HRV.Results In 2 206 cases,total RSV positive rate was 19.90％ (439/2 206 cases) and simple RSV infection positive was detected in 399 cases.Total HRV positive rate was 14.14％ (312/2 206 cases),in which simple HRV infection positive was detected in 250 cases and the detection rate of RSV was significantly higher than that of HRV(x2 =25.88,P ＜0.05).The incidence rate of wheezing in simple RSV infection was 68.17％ (272/399 cases),which was significantly higher than that of simple HRV infection (42.80％,107/250 cases) (x2 =11.174,P ＜ 0.05).RSV infection was frequent from November to February of the next year in which the detection rate in December was highest with the proportion of 50.00％ (99/198 cases) while the rate in June was only 0.57％ (1/175 cases).The detection rate of HRV was 22.86％ (40/175 cases),20.47％ (35/171 cases) and 20.33％ (25/123 cases) in June,July and September respectively.The detection rate of HRV was lower during December to February of the next year.In January,the detection rate was only 4.68％ (11/235 cases),which was the lowest in the whole year.The detection rates of RSV were 33.33％ (4/12 cases),25.21％ (118/468 cases),23.46％ (84/358 cases) and 23.81％ (60/252 cases) in the age group of 28 d-1 month,＞ 1-2 month,＞ 2-3 month and ＞ 3-4 months respectively.Up to the age of 4 months old,the detection rate decreased gradually,and with the increase of age and the detection rate in ＞ 7-8 month group was only 10.96％ (16/146 cases).The detection rate of HRV was 0 (0/12)and 9.40％ (44/468 cases) in the age group of 28 d1 month,＞ 1-2 month,respectively.After 2 months age old,the detection rate fluctuation ranged from 13.22％ to 16.67％.The incidence rate of severe RSV infection was 12.30％ (54/439 cases) and the incidence rate of severe HRV infection was 5.13％ (16/312 cases).Increased respiratory rate was more common in patients with severe RSV infection while severe HRV infection in infants were accompanied by multiple lobar involvement.After RSV infection,the incidence rate of pulmonary function damage was 89.03％ (276/310 cases).After HRV infection,89.27％ (183/205 cases)of the infants suffered from pulmonary function damage.Both RSV and HRV infection might cause pulmonary function damage.Conclusions RSV and HRV are the major pathogens in infants of Suzhou areas.The incidence of RSV-induced wheezing is significantly higher than that of HRV.RSV is detected positive mainly in winter and early spring and the infants within 4-month old are susceptible population.HRV is detected positive mainly in June,July and September and the infants older than 2 months are susceptible population.The incidence of severe RSV infection is significantly higher than that of HRV.Severe RSV infection may cause increased respiratory rate and severe HRV infection mainly cause multiple lobar involvement.RSV and HRV infection may cause pulmonary function damage.

BACKGROUND:Hepatocyte transplantation has attracted more and more attention as a therapeutic measure for liver failure and genetic metabolic liver diseases.OBJECTIVE:TO evaluate the efficacy and safety of human hepatocyte transplantation in treating hepatitis B related liver failure in one case by a 2-year follow-up.DESIGN:A case-report of 2-year follow-up.SETTING:No.9 Department of Infectious Diseases,Bioengineering Research Room,the 302 Hospital of Chinese PLA.PARTICI PANT:One inpatient with hepatitis B related liver failure was selected from the 302 Hospital of Chinese PLA.and she was diagnosed according the laboratory tests.The transplanted hepatocytes were originated frOm the healthy liver of a 24-year-old man,who had signed the protocol for liver donation before death.METHODS:The hepatocyte transplantation was completed in the Department of Radiology,the 302 Hospital of Chinese PLA in December 2004.Liver was isolated to obtain human primary hepatocytes, and then cryopreserved.The hepatocytes were transplanted into recipient spleen via femoral vein after resuscitation.The clinical symptoms,changes of blood biochemical indexes,and changes of spleen MRI signals were observed before and after operation.The patient was reexamined every half a year after operation, including liver function, blood coagulation function,B-mode ultrasonography,gastroscopy and MRI,and she was followed up for 2 years. MAIN OUTCOME MEASURES:Liver function,blood coagulation function, imaging indexes, immunological indexes,complication and rejection.RESULTS:①Totally(1-2)×1010 hepatocytes were harvested,and the viability of rewarmed hepatocytes was 60%,and finally 2×109 hepatocytes were transplanted.②Two months later,the clinical symptoms of the recipient were obviously ameliorated,and serum bilirubin and aspartate aminotransferase(AST)were obviously decreased,while prothrombin activity was markedly increased.20 months later,the MRI results showed that there was hepatocyte image in spleen.Two years after operation.the total bilirubin level was 20 μmol/L,direct bilirubin level was 7 μmol/L, alanine aminotransferase was 416.75 nkat/L,AST was 533.44 nkat/L,albumin was 37 g/L,prothrombin activity was 90%,which were all obviously ameliorated as compared with those before operation(474.5 μmol/L,340.3 μmol/L,400.08 nkat/L,1 200.24 nkat/L,38 g/L,25%).The patient left the hospital 2 months later and could do light-burdened job.No complications of hydroperitonia and liver function failure, etc.were observed,and no rejection occurred.Several reexaminations by B-mode ultrasonography all indicated the further aggravations of liver cirrhosis and esophageal varices.She was admitted to hospital for twice because of esophageal varices bleeding,and cured by endoscopic variceal sclerosis therapy.CONCLUSION:Hepatocyte transplantation can ameliorate liver function without rejection,but it cannot relieve portal hypertension.

Objective To study the epidemiological and clinical features of viral pathogen of acute respiratory tract infection(ARTI)in children in Suzhou. To provide the evidence-based medicine for early warning,diagnosis and treatment of respiratory tract infection in children. Methods The study population comprised of all the children ranging from 28 days to 15 years old with ARTI admitted to Department of Respiration,the Childrenˊs Hospital Affiliated to Soo-chow University,between January 2006 and December 2015. The study cases were divided into 4 age groups based on ages(28 days to <1 year old,1- <3 years old,3- <5 years old and ≥5 years old). The nasopharyngeal aspirates and medical history were obtained by qualified medical personnel. Seven kinds of common respiratory virus were detec-ted by using the direct immunofluorescence. Mycoplasma pneumonia(MP),chlamydia pneumonia(CP)and human bo-cavirus(hBoV)were detected by adopting fluorescence quantitative PCR. Human rhinovirus(HRV)and human metap-neumovirus(hMPV)were detected by means of reverse transcriptionpolymerase chain reaction( RT -PCR). Sputum was cultured for bacteria. Results In 21 624 cases,17 618 cases were detected,and the positive detection rate of virus was 30. 74%(5 416 cases). The highest detection rate of virus was 44. 25%(881/1 991 cases)in 2011 in the 10 years. There were 3 317 cases of single virus pathogen infection,accounting for 18. 83%. Among them,respiratory syn-cytial virus(RSV)was the first with 9. 42%(1 660/17 618 cases),followed by HRV(5. 17%,278/5 372 cases), hBoV(3. 12%,351/11 263 cases),Pinf 3(2. 17%,383/17 618 cases),and the lowest rate was Pinf 2 with 0. 05%(8/17 618 cases). There were 2 099 cases of overlap infection,accounting for 11. 91%. The virus combined with bacte-rial infection accounted for 7. 86%(1 384/17 618 cases),which had the most positive cases. With the increase of age, the single virus pathogen infection rate gradually dropped. The highest detection rate was 22. 82%(1 976/8 656 cases) in the group between 28 days and <1 year old,and the lowest detection rate was 7. 81%(159/2 035 cases)of children over 5 years of age. The virus positive rate of the four seasons was 18. 28%(833/4 556 cases),13. 86%(625/4 508 cases),5. 91%(684/4 298 cases),27. 68%(1 178/4 256 cases),respectively,and detection rate in winter was the highest,and the least in summer. In the single virus infection,bronchiolitis had the highest percentage(36. 32%,25/69 cases),and the least rate was 5. 83%(63/1 080 cases)of lobar pneumonia. Conclusions Monitored for 10 years of the virus respiratory infection in Suzhou area,RSV,HRV and hBoV turned out to play an important role in respiratory tract infection in children. The children less than 3 years old are the most susceptible to virus infection. The epidemic season of each virus is different and characteristic.

Medical humanistic spirit should be reflected in the whole process and the all-round of medical care.However,limited to the current cognition and condition,medical humanities in clinical practice are still not comprehensive and universal.By reading the existing literature and analyzing,combining with the own work experi-ence at the same time,this paper agreed that aiming at the special disease of cancer,the humanistic practice in the treatment of nursing had its own importance and its particularity,and it should recognize the disease and confront the patients and their families from the perspective of the whole person and omni-direction,make traditional clini-cal humanities extend to the family and extend humanistic care to family members of the patients.In this practice,medical staff can cultivate the consciousness of awing the life and respecting the patient in specific operations,es-tablish the sense of integration and improve their spiritual realm,so as to continuously improve their comprehensive quality in nursing service.

Objective To explore the function of extended nursing combined with PDSA circle and based on positivepsychology in improving self-efficacy of patients with colostomy. Methods A total of 80 patients with colostomy match the inclusion and exclusion standard from December 2014 to December 2015, were divided into the control group and the intervention group according to the order of discharge date.Patients in the control group received conventional extended nursing, while patients in the intervention group received extended nursing combined with PDSA circle and based on positivepsychology, and interventions were conducted a week, a month, two months, three months after discharge. Patients were evaluated by stoma related self-efficacy questionnaire before the intervention, after the fourth intervention and 3 months after intervention ( T0 , T1 ,T2 ) . Results There were significant differences in the dimensions, independent items and total scores of stoma related self-efficacy between two groups at different time after intervention except the item of confidence in sexual life (P<0.05), and they also had interactive effect (P<0.05).There was significant difference in the main effect of intervention time for the single item “confidence in sexual life” (P<0.05). In the intervention group, the scores of dimensions, independent items and total scores of stoma related self-efficiency after intervention and three months after intervention were significantly higher than those before intervention. Conclusions The extended nursing combined with PDSA circle and based on positivepsychology can improve the self-efficacy of patients with colostomy.

OBJECTIVETo clone the unknown gene of hepatocyte protein interacting with hepatitis C virus core protein.

METHODSUsing the yeast dual hybrid system 3, bait plasmids of hepatitis C virus core were constructed. After identifying hepatitis C virus core protein that could stably expressed in AH109 yeast strains, we performed yeast two hybrid by mating AH109 with Y187 that transformed with liver cDNA library plasmids pACT2 and then plated on quadrople dropout (QDO) medium and assayed for alpha-gal activity. The genes of yeast colonies that could grow on QDO and had alpha-gal activity were sequenced.

RESULTSAmong the 30 positive colonies, we blasted the gene of the sixth colony; we coined human hepatitis C virus binding protein 6(Hu Hcbp6) with Genbank, realized that the Hu Hcbp6 shares as much as 98% homology with two cDNA without knowing functions. We have proved that Hu Hcbp6 could interact with hepatitis C virus core protein.

CONCLUSIONSHepatitis C virus core binding protein (Hu Hcbp 6 Genbank number: AY032594) was successfully cloned and identified. The study partly paved the way for investigating physiological function of the Hu Hcbp6.

Cloning, Molecular ; DNA, Complementary ; genetics ; DNA-Binding Proteins ; genetics ; Hepacivirus ; Humans ; Molecular Sequence Data ; Plasmids ; Sequence Analysis, DNA ; Transfection ; Two-Hybrid System Techniques ; Viral Core Proteins ; genetics ; metabolism ; Yeasts ; genetics

Objective:To evaluate the diagnostic efficacy and practicality of the Jaundice color card (JCard) as a screening tool for neonatal jaundice.Methods:Following the standards for reporting of diagnostic accuracy studies (STARD) statement, a multicenter prospective study was conducted in 9 hospitals in China from October 2019 to September 2021. A total of 845 newborns who were admitted to the hospital or outpatient department for liver function testing due to their own diseases. The inclusion criteria were a gestational age of ≥35 weeks, a birth weight of ≥2 000 g, and an age of ≤28 days. The neonate′s parents used the JCard to measure jaundice at the neonate′s cheek. Within 2 hours of the JCard measurement, transcutaneous bilirubin (TcB) was measured with a JH20-1B device and total serum bilirubin (TSB) was detected. The Pearson′s correlation analysis, Bland-Altman plots and the receiver operating characteristic (ROC) curve were used for statistic analysis.Results:Out of the 854 newborns, 445 were male and 409 were female; 46 were born at 35-36 weeks of gestational age and 808 were born at ≥37 weeks of gestational age. Additionally, 432 cases were aged 0-3 days, 236 cases were aged 4-7 days, and 186 cases were aged 8-28 days. The TSB level was (227.4±89.6) μmol/L, with a range of 23.7-717.0 μmol/L. The JCard level was (221.4±77.0) μmol/L and the TcB level was (252.5±76.0) μmol/L. Both the JCard and TcB values showed good correlation ( r=0.77 and 0.80, respectively) and agreements (96.0% (820/854) and 95.2% (813/854) of samples fell within the 95% limits of agreement, respectively) with TSB. The JCard value of 12 had a sensitivity of 0.93 and specificity of 0.75 for identifying a TSB ≥205.2?μmol/L, and a sensitivity of 1.00 and specificity of 0.35 for identifying a TSB ≥342.0?μmol/L. The TcB value of 205.2?μmol/L had a sensitivity of 0.97 and specificity of 0.60 for identifying TSB levels of 205.2 μmol/L, and a sensitivity of 1.00 and specificity of 0.26 for identifying TSB levels of 342.0 μmol/L. The areas under the ROC curve (AUC) of JCard for identifying TSB levels of 153.9, 205.2, 256.5, and 342.0 μmol/L were 0.96, 0.92, 0.83, and 0.83, respectively. The AUC of TcB were 0.94, 0.91, 0.86, and 0.87, respectively. There were both no significant differences between the AUC of JCard and TcB in identifying TSB levels of 153.9 and 205.2 μmol/L (both P>0.05). However, the AUC of JCard were both lower than those of TcB in identifying TSB levels of 256.5 and 342.0 μmol/L (both P<0.05). Conclusions:JCard can be used to classify different levels of bilirubin, but its diagnostic efficacy decreases with increasing bilirubin levels. When TSB level are ≤205.2 μmol/L, its diagnostic efficacy is equivalent to that of the JH20-1B. To prevent the misdiagnosis of severe jaundice, it is recommended that parents use a low JCard score, such as 12, to identify severe hyperbilirubinemia (TSB ≥342.0 μmol/L).