Objective To clone and express human autoantigen Sm B'in methylotrophie yeast Pichia Pagtoris.Methods The gene Sm B' was cloned bv PCR The PCR product wag inserted into the vector pPIC9k.The recombinant plasmid pPIC9k.Sm B' was transformed into yeast Sm D1168 by electroporation.The positive clones were screened in MD plates.The high copy number transformants were rapidly selected by using G418 and were induced by methan01.Supematants after induction were analyzed by SDS-PAGE and western blot.Sera collected from thirty patients with SLE.thirty patients with mixed connective tissue disease(MCTD)and thirty healthy volunteers were detected by immunodot and immunoblot.Results The PCR product wag about 700 bD in size which Wag in accordance with predicted 657 bp.The pPIC9k-Sm B'showed the same seqencing result with GenBank's report and restriction enzyme analysis confirmed our prediction.The pPIC9k-Sm B' positive clone produced a 32 000 protein which had natural immunogenicitv of human autoantigen Sm B'by SDS-PAGE and western blot.The positive rate of immunodot and IBT were 46.7%(42/90)and 51.1%(46/90),respectively.The agreement between immunodot and IBT was very close(Kappa value=0.911 2,P＜0.01).Conclusion Successfully cloning and expression of human autoantigen Sm B' in methylotmphic yeast Pichia Pagtoris hid a foundation for further research work.

Chronic hepatitis C virus ( HCV) infection has been reported in association with several extrahepatic complications, among which mixed cryoglobulinemia is the most documented. Clinical manifestations of HCV related cryoglobulinemia include skin lesions, glomerulonephritis, peripheral neuropathy, and widespread vasculitis.The therapy may be directed to eradicate HCV infections, suppress B-cell clonal expansion and cryoglobulin production, and ameliorate symptoms.This paper reviews the research progress on pathogenesis, clinical manifestations, diagnosis and treatment of HCV related cryoglobulinemia.

The aim of this study is to establish a multiplex polymerase chain raction (PCR) to identify of four kinds of laboratory animal pathogens: Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae and Klebsiella pneumoniae.Methods Specific primers were designed based on GenBank data.The multiplex PCR system was established through optimization of multiple PCR and detection of its specificity and sensitivity.This technique was used to test artificially infected samples and tracheal secretions of experimental animals (rat, mouse, guinea pig, rabbit, hamster), and comparing the detection results by this method and traditional detection test.Results Target bands of Pasteurella multocida (356 bp), Bordetella bronchiseptica (237 bp), Mycoplasma pneumoniae (266 bp), and Klebsiella pneumoniae (142 bp) were obtained, with a detection sensitivity of Klebsiella pneumoniae of 10 pg, and that of Pasteurella multocida, Bordetella bronchiseptica and Mycoplasma pneumoniae of 1 pg by this newly developed multiplex PCR assay.No target bands were observed from the non-specific pathogens of artificially infected samples.The tracheal secretions taken from 45 experimental animals (mice and rabbits) were tested with this new PCR assay, among which 15 cases of Klebsiella pneumonia and 9 cases of Pasteurella multocida were detected as positive, while all the results of traditional method and serological test were negative.Conclusions A simple, rapid, specific and highly sensitive multiplex PCR system has been successfully established.It is valuable for detection of Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae, and Klebsiella pneumoniae in laboratory animals.

Objective To investigate the efficacy of micrornas(miR)-27B-3P combined with miR-215 in predicting the recurrence of early gastric cancer after endoscopic submucosal dissection(ESD).Methods A total of 122 patients with early gastric cancer admitted to the hospital from January 2021 to February 2022 were selected.All patients received ESD surgery and were followed up for 10～15 months,average(12.57±1.83)months.Patients with early gastric cancer were divided into recurrence group and non-recurrence group according to whether they had recurrence after ESD surgery.The relative expression levels of miR-27b-3p and miR-215 and clinical data of the two groups were compared to analyze the factors affecting the recurrence of early gastric cancer patients after ESD surgery,and analyze the relative expression levels of miR-27b-3p and miR-215 and the predictive value of their combination for the recurrence of early gastric cancer patients after ESD surgery.Results After 10～15 months,average(12.57±1.83)months of follow-up,3 cases of 122 patients were lost to follow-up,the follow-up rate was 97.54％,of which 12 cases relapsed,and the remaining 107 cases did not relapse.The miR-27b-3p in the relapsed group was lower than that in the non-relapsed group(P＜0.05),and the relative expression of miR-215 in the relapsed group was higher than that in the non-relapsed group(P＜0.05).The proportion of patients with lesion size ≥3 cm and infiltration depth of SM2-SM3 in recurrence group was higher than that in non-recurrence group(P＜0.05),and the proportion of patients with R0 resection in recurrence group was lower than that in non-recurrence group(P＜0.05).Logistic multivariate regression analysis showed that the lesion size was ≥3 cm and the depth of invasion was SM2-SM3,the relative expression level of miR-27b-3p,the relative expression level of miR-215 were the influential factors for recurrence of early gastric cancer patients after ESD operation(P＜0.05).The relative expression levels of miR-27b-3p and miR-215 and their combined prediction of the area under the curve(AUC)of ESD postoperative recurrence in early gastric cancer patients were 0.783,0.845 and 0.935,respectively.Conclusion miR-27b-3p and miR-215 can be used to predict the recurrence risk of early gastric cancer patients after ESD surgery,and the combination of Mir-27B-3P and Mir-215 has higher predictive value.

Cardiovascular surgery is a type of high-risk surgery for pressure ulcers. This review summarizes the risk factors of pressure ulcers in patients undergoing cardiovascular surgery, including preoperative, intraoperative and postoperative risk factors. At the same time, it points out that in the present study, there is a dispute about whether the pressure sore is associated with the time of cardiopulmonary bypass and the change of body temperature in patients undergoing cardiovascular surgery. In addition, here is a lack of related research on whether the anticoagulant of protamine has an effect on pressure sores during the operation. This article aims to provide a basis for the clinical observation of the risk factors of pressure sores in cardiovascular surgery.

Objective To retrieve and integrate relevant evidence on the prevention and nursing of neonatal intraoperative acquired pressure and provide a reference basis for nursing practice.Methods According to the"6S"model retrieval strategy,we searched for published articles including the following topic words,such as prevention,assessment,management,and nursing care of neonatal intra-operative pressure injury from establishment of databases until August 2,2023.We got the related references from clinical decision-making websites,guidelines network,professional association websites,domestic and foreign databases.Results Totally 14 articles were included,including 1 clinical decision support,4 clinical guidelines,3 evidence summaries,2 systematic reviews and 4 expert consensuses.Finally,31 pieces of best evidence were summarized from 5 aspects of risk factors,risk assessment,observation and preventive measures,nursing measures after the occurrence of pressure injury,education and training.Conclusion Multidisciplinary collaborative participation based on the obtained evidence is an effective strategy,which can be applied to the prevention and nursing care of neonatal intraoperative acquired pressure injury.Appropriate preventive and nursing programs should be made according to the actual situation and clinical environment,so as to reduce the incidence of neonatal intraoperative acquired pressure injury and to ensure the safety of neonatal perioperative nursing care.

Objective To investigate effect of let-7a gene over-expression on apoptosis of gastric cancer cell lines SGC-7901.Methods The stable let-7a gene over-expressing SGC-7901 cells, SGC-7901/let-7a cells, were established using shRNA lentiviral vector methods. Real-time RT-PCR analysis was used to evaluate the expression level of let-7a mRNA. Cells apoptosis was assessed by flow cytometry.Results RT-PCR analysis revealed let-7a expression in SGC-7901/let-7a cells was significantly increased. Cellular apoptosis assay showed that over-expression of let-7a could increase apoptosis of SGC-7901 cells (P=0.002).Conclusion Up-regulating let-7a expression promoted apoptosis in SGC-7901 cells.
Apoptosis ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; MicroRNAs ; biosynthesis ; genetics ; RNA, Neoplasm ; biosynthesis ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Up-Regulation

Objective To investigate the effects of gemstone energy spectrum CT atomic number method and infrared spec-troscopy for analyzing the composition of urinary calculi and to compare their values in qualitative diagnosis of urinary calculi .Meth-ods Two hundreds and sixty cases of urinary tract stones were performed the gemstone spectrum CT urinary scanning and the stone composition was identified by atomic number method .After removing stone ,the stone composition analyzed by infrared spec-troscopy served as the gold standard .Then the consistency identified by the two methods was analyzed .Results The Kappa consis-tency test results showed that the two kinds of method for identifying stone type had good consistency (Kappa=0 .787 ,P<0 .01) . The paired chi square test results showed that the difference of the two methods for identifying the stone type had no statistical sig-nificance(χ2 =6 .581 ,P=0 .254) .The stone crystal composition types measured by gemstone energy spectrum CT atomic number method were less than those measured by infrared spectroscopy .The precise quantification of the stones with different crystal struc-tures was not as accurate as that of infrared spectroscopy (calcium oxalate monohydrate and calcium oxalate dihydrate ) .Conclusion The two methods for analyzing theurinary stone composition all have clinical significance ,the stone analysis method should be se-lected according to the actual situation .

Objective To study the epidemiology of hand,foot,and mouth disease (HFMD) and the spectrum of serotypes in the other enterovirus (EV) (non-EV-A71 and non-Coxsaekievirus group A 16,CV-A 16) from 2016 to 2017 in Guangzhou,to provide the basis for its treatment,prevention and control.Methods Enteroviruses universal type,EV-A71 and CV-A16 were detected by real time reverse transeription-polymerase chain reaction in the specimens from HFMD suspected patients from 2016 to 2017.The positive specimens of non-EV-A71 and non-CV-A16 were amplified and sequenced based on 5'-untranslated region (UTR) region.The spectrum of serotypes was analyzed with BLAST in NCBI on the basis of 5'-UTR region.Results A total of 25779 specimens from HFMD patients were collected during 2016-2017,16 300 (63.23 ％) of which were positive.The positive rates of EV-A71,CV-A16,non-EV-A71 and non-CV-A16 were 4.57％ (1 178/25 779),12.70％ (3 274/25 779) and 45.96％ (11 848/25779),respectively.The average positive rate of non-EV-A71 and non-CV-A16 in 2017 was 55.68％,which was higher than that in 2016.Sequence analysis showed that there were 16 genotypes in 95 non-EV-A71 and non-CV-A16 positive specimen,including CV-A6,CV-A10,CV-A4,CV-A2,CV-A8,CV-A12,CV-A9,Coxsakievirus B5 (CV-B5),CV-B2,CV-B4,CV-B3,Echovirus 1 (E1),E16,E30,E2 and E18.CV-A6 (26.32％),and CV-A10 (15.79％) were the most common genotypes,followed by CV-A4 (6.32％)、CV-A8(4.21％),and CV-A2 (4.21％).Conclusions The infection rate of EV-A71 is very low during 2016-2017.From April to July 2016,there is a small peak of CV-A16 infection.The non-EV-A71 and non-CV-A16 enterovirus becomes the main causative agent of HFMD during 2016 to 2017.CV-A6 and CV-A10 are the most prevalent pathogens of non-EV-A71 and non-CV-A16 enterovirus.Research and monitoring of CV-A6,CV-A10 as the main non-EV-A71and non-CV-A16 virus should be strengthened.

Objective:To investigate the epidemiology of pathogens of acute respiratory tract infection (ARTI) in children in Guangzhou area.Methods:A total of 13 610 hospitalized children with ARTI in Guangzhou Women and Children′s Medical Center from January 2018 to December 2021 were enrolled. Throat swab specimens were collected, and fluorescent quantitative polymerase chain reaction (PCR) was performed to detect 11 respiratory pathogens, including respiratory syncytial virus (RSV), adenovirus (ADV), parainfluenza virus (PIV), human rhinovirus (HRV), human bocavirus (HBoV), human metapneumovirus (HMPV), enterovirus (EV), influenza A virus (IFA), influenza B virus (IFB), Mycoplasma pneumoniae (MP) and Chlamydia pneumoniae (CP). Grouping according to age (< one year group, one to < three years group, three to < six years group, six to 14 years group) and season. Chi-square test was used for statistical analysis. Results:At least one pathogen was detected in 6 331 cases among 13 610 patients, and the overall positive rate was 46.52%. The detection rates from high to low were as follows: RSV (13.75%(1 872/13 610)), ADV (4.82%(656/13 610)), PIV (4.82%(656/13 610)), MP (4.54%(618/13 610)), HRV (3.39%(462/13 610)), HBoV (2.64%(359/13 610)), HMPV (2.59%(352/13 610)), EV (1.76%(239/13 610)), IFA (1.29%(176/13 610)), IFB (0.90%(122/13 610)) and CP (0.30%(41/13 610)). The positive rate of viral detection showed significant differences among different age groups ( χ2=49.91, P<0.001), and the highest positive rate was in the age group of one to