Objective To construct the recombinant streptolysin O antigen(SLO) prokaryotic expression plasmid and establish its best expression condition in Escherichia coli .Methods The DNA fragment encoding SLO was amplified from streptococcal ge-nomic DNA template by PCR ,and then incorporated into pET-32a(+ ) vector to construct pET-32a(+ )-SLO recombinant plas-mid .pET-32a(+ )-SLO was transformed into Escherichia coli BL21(DE3) and SLO protein was expressed and purified by isopro-pyl-β-D-thiogalactoside(IPTG)-induction and auto-induction ,respectively .Results The results of DNA electrophoresis and DNA sequencing showed that pET-32a(+ )-SLO recombinant plasmid was constructed successfully .When IPTG at different concentra-tion was used ,SLO expressed as inclusion body and its expression efficiency was low .Under auto-induction condition ,SLO ex-pressed as partly soluble manner ,and its expression efficiency increased .Conclusion The prokaryotic expression plasmid pET-32a (+ )-SLO is constructed successfully and the best condition for SLO expression and purification from Escherichia coli culture is es-tablished ,which lay the foundation for further basic and clinical application research with SLO .

Objective To explore the psychological stress of young nurses facing patients' death. Methods Young nurses in emergency department, ICU and hospice care ward (96 cases, research group) and young nurses in other departments(94 cases,control group)from 8 general hospitals were investigated by self-rating anxiety scale (SAS)and serf-rating depression scale (SDS).The results underwent comparison and analysis. Results SAS and SDS score,incidence of anxiety and depression in the research group were higher than those of the control group (P ＜ 0.05). Conclusions Young nurses who face patients' death frequently have high incidence rate of psychological stress.They show low psychological health level which deserves the close attention of nursing managers.

BACKGROUND:Studies on the proteomics contribute not only to exploring the laws governing life activities, but also to elucidating the pathogenesis of a variety of diseases to find the treatment strategies. OBJECTIVE:To summarize the application of proteomics in the plateau medical research. METHODS:A computer-based online search was conducted in PubMed and Wanfang databases from 1995 to 2015 to screen the relevant literatures using the key words“proteomics, medical research, plateau medicine”in English and Chinese, respectively. A total of approximately 200 English and 60 Chinese relevant literatures were selected and the 59 eligible literatures were included after screening final y. RESULTS AND CONCLUSION:Currently, the human genome has been decoded through the studies on exploring the proteomics changes under the pathological conditions and the underlying mechanisms. Construction of physiological and pathological mapping based on the human proteome contributes to revealing the novel treatment targets, diagnostic markers, and drugs for the prevention and treatment of diseases. Proteomics has become the frontier and hot research field both at home and abroad, including al the proteins expressed in the tissue, cel or organism, and becomes a bridge between the genome and clinical application.

Objective To investigate the influence of different volumes of ropivacaine with isod-osage on diaphragmatic excursion following ultrasound-guided interscalene brachial plexus block. Methods Sixty ASA Ⅰ-Ⅱ patients scheduled for right ulnoradial fracture internal fixation removal were randomized into group A (0.5% ropivacaine 20 ml)and group B (0.75% ropivacaine 13.3 ml) (n=30,each).Diaphragmatic excursion were evaluated by M type ultrasound before,1 5 min and 30 min after drug injection. The degree and duration of sensory and motor block were recorded. Results The anesthetic effect and maintain time had no significant difference between groups;Com-pared with group A,group B had lower incidence of diaphragmatic paralysis (P <0.05).There were 28 (93%)and 22 (73%)cases of patients in group A and group B respectively showed reduction of diaphragmatic excursion more than 50% or even complete paralysis 30 min after drug injection (P <0.05).Conclusion Ultrasound-guided interscalene brachial plexus with 0.5% ropivacaine 20 ml or 0.75% ropivacaine 13.3 ml both can provide excellent block.Compared with 0.75% ropivacaine 13.3 ml,0.5% ropivacaine 20 ml is more likely to lead to diaphragmatic paralysis.

Objective To explore the relationship between the immune function of cellin peripheral blood with the virureplication and hepatitiviru(HCV)-cAg expression in the patientwith chronihepatiti(CHC) .MethodPeripheral blood lymphocytesubpopulation ,HCV-Rnand HCV core antigen (HCV-cAg) in 63 healthy people undergoing the physical exami-nation (control group) and 85 caseof CHC(Chgroup) were analyzed by the flow cytometry ,real-time Pcand ELIS,respec-tively .ResultThe percentageof total cell,T4 cell,T8 cell,double negative cell(DN) and double positive cell(DP) in the Chgroup were (67 .37 ± 10 .43)% ,(37 .11 ± 10 .28)% ,(21 .63 ± 8 .87)% ,(7 .80 ± 4 .57)% and (0 .20 ± 0 .29)% , respectively ,the absolute contentwere in turn (0 .70 ± 0 .44) × 109/L ,(0 .37 ± 0 .22) × 109/L ,(0 .22 ± 0 .17) × 109/L ,(0 .08 ± 0.06)×109/Land(0.19±0.68)×107/L,respectively.TheratioofT4/T8was(2.18±1.26)% .Theresultsindicatedthatthe percentage of T8 cellin the Chpatientwadecreased obviously (P＜0 .01) ,which resulted in the ratio of T4/T8 raising(P＜0 .05);meanwhile ,the absolute contentof the total cell,T4 cell,T8 celland Dnwere all decreased obviously (P＜0 .05);moreove,the percentage of T4 celland Dnin the patientwith HCV-Rnpositive and HCV-cAg positive wasignificantly in-creased (P＜0 .05) .Conclusion When HCV replicating in the patientwith CHC,the T lymphocyte subpopulation haobviouab-normity .The low immune function or immune tolerance ofT cells may be the important cause of recurrence and uncurability of CHC.

Objective To evaluate the diagnostic value and safety of domestic electromagnetic navigation bronchoscopy (ENB) guided transbronchial lung biopsy (TBLB) for peripheral pulmonary lesions.Methods Sixty-four patients with peripheral pulmonary lesions shown by thoracic CT in Zhongshan Hospital,Fudan University between Jul.and Dec.,2014 were collected.The patients were randomly assigned to test group (underwent ENB in combination with X-ray guided TBLB) and control group (underwent X-ray guided TBLB).The final diagnosis was confirmed by pathologic examination of surgically removed lesions or by 24 months clinical follow-up.The operative time as well as the intraoperative and postoperative complications were also recorded.Results Sixty-four patients had 70 peripheral pulmonary lesions.There was no difference in age,sex,the lesion size or location between the two groups.Pathology results showed that the diagnostic yield of test group and control group were 88.6％ and 62.9％,respectively,with statistical significance (P =0.012).Subgroup analysis showed that if the lesion's diameter was ≤2 cm,the diagnosis yield of test group was higher than control group (66.7％ vs.20.0％,P =0.266);if the lesion was ＞2 and ≤3 cm,the diagnosis yield of test group and control group were 100 ％ and 81.8 ％,respectively (P =0.485).But if the lesion was＞3 cm,the diagnostic yield of the 2 group was significantly different (94.4 ％ in test group,63.1％ in control group,P =0.042).Mean operation duration of the 2 group was (966 ± 372)s and (1 040 ± 470) s,respectively,with no statistical difference (P =0.600).However,there was statistical difference between the 2 groups on the X-ray time needed to find the pulmonary lessions [(7.0 ± 4.8)s vs.(37.0 ± 37.5) s,P =0.008).There was no pneumothoraxes and excessive bleeding in patients undergoing ENB.Conclusions Compared with X-ray guide TBLB,ENB guided TBLB for peripheral pulmonary lesions has a certain degree of security,and has superiority in reducing the X-ray time required to find the lesion and improving diagnostic yield especially when the lesion's diameter was ＞3 cm.

In this study, the current status for breast diseases in a region with high-incidence of cervical cancer were epidemiologically investigated. From March to August, 2009, 17618 women, from Wufeng area of Hubei province, China, were recruited to screen breast diseases by using breast infrared diagnostic apparatus. Other diagnostic methods, such as B-mode ultrasound, X-ray mammography, needle biopsy and pathological examination were, if necessary, used to further confirm the diagnosis. The screening showed that 5990 of 17618 cases (34.00%) had breast diseases, 5843 (33.16%) had mammary gland hyperplasia, 48 (0.27%) had breast fibroadenoma, 11 (0.06%) had breast carcinoma, and 88 (0.50%) had other breast diseases. The peak morbidity of breast cancer was found in the women aged 50-60 ages. The morbidity of breast cancer was significantly increased in women elder than or equal to 50 years old (n=8, 0.157%) in comparison with that in the subjects younger than 50 years old (n=3, 0.024%) (u=2.327, P<0.05). It was shown that the occurrence of breast diseases was concentrated in women aged 20-40 years, while the total morbidity reached its peak at the age of 30 years and then decreased sharply after age of 40. Compared with the patients elder than or equal to 40 years old (n=3289, 27.46%), the morbidity rate of breast diseases was significantly increased in women less than 40 years old (2648 cases, 47.18%; P<0.001). However, there was no significant difference in the morbidity of breast diseases between the age group of 20-29 years and that of 30-39 years (P=0.453), and both of them were high. There was no significant association between the morbidity of breast diseases and cervical cancer. Since the morbidity of breast diseases was higher among young women, more attention should be paid to the screening of breast diseases among young women for early diagnosis.

Objective:To construct and validate a predictive model for the risk of excessive blood loss in pa-tients with ruptured tubal pregnancy,and to provide a basis and tool for the assessment of changes in the condi-tion of patients with ruptured tubal pregnancy.Methods:Clinical data of inpatients with ruptured tubal pregnancy from January 2014 to July 2021 were retrospectively analyzed,who underwent surgical treatment in the Depart-ment of Gynecology,Dongguan Maternal and Child Health Hospital.The pelvic blood volume was categorized into excessive blood loss and non-excessive blood loss groups based on whether the amount of pelvic blood was found to be≥750 ml intraoperatively.Factors influencing the occurrence of excessive blood loss were screened and modeled by univariate analysis,Lasso regression,and multi-factor Logistic stepwise regression.The area un-der the subject working characteristic curve(AUC)was used to evaluate the discrimination of the predictive mod-el,the model's consistency was evaluated by calibration curve and goodness-of-fit test,and the clinical utility of the model was evaluated and validated by the decision analysis curve.Finally,column line plots were drawn.Results:①A total of 386 patients with ruptured tubal pregnancy were included,of whom 124(32.12%)had blood loss≥750 ml.②The optimal predictors for predicting concomitant blood loss in patients with ruptured tubal preg-nancy were screened,including:days of abdominal pain,dizziness,pallor,fatigue,the maximum diameter of para-metrial mass,human chorionic gonadotropin(β-hCG),and hemoglobin(Hb)and the model and the column line graphswere constructed accordingly.③The prediction model AUC was 0.827(95%CI 0.781-0.873);the cut-off value was 0.391,at which point the specificity and sensitivity were 68.55%and 84.35%,respectively,and the AUC validated within the model by resampling was 0.804.Clinical decision curves showed that the threshold probability intervals for the maximum net benefit values ranged from 8.5%-97%,respectively.Conclusions:The constructed prediction model was validated to suggest good discriminatory efficacy and degree of consistency.As a tool,it has clinical application value in predicting the risk of hemorrhage in patients with ruptured tubal pregnan-cy.It can help to determine the occurrence of adverse events such as hemorrhagic shock at an early stage and improve the success rate of rescue treatment.

OBJECTIVETo study the severe acute respiratory syndrome (SARS)-associated coronavirus genotype and its characteristics.

METHODSA SARS-associated coronavirus isolate named ZJ01 was obtained from throat swab samples taken from a patient in Hangzhou, Zhejing province. The complete genome sequence of ZJ01 consisted of 29,715 bp (GenBank accession: AY297028, version: gi: 30910859). Seventeen SARS-associated coronavirus genome sequences in GenBank were compared to analyze the common sequence variations and the probability of co-occurrence of multiple polymorphisms or mutations. Phylogenetic analysis of those sequences was done.

RESULTSBy bioinformatics processing and analysis, the 5 loci nucleotides at ZJ01 genome were found being T, T, G, T and T, respectively. Compared with other SARS-associated coronavirus genomes in the GenBank database, an A/G mutation was detected besides the other 4 mutation loci (C:G:C:C/T:T:T:T) involved in this genetic signature. Therefore a new definition was put forward according to the 5 mutation loci. SARS-associated coronavirus strains would be grouped into two genotypes (C:G:A:C:C/T:T:G:T:T), and abbreviated as SARS coronavirus C genotype and T genotype. On the basis of this new definition, the ZJ01 isolate belongs to SARS-associated coronavirus T genotype, first discovered and reported in mainland China. Phylogenetic analysis of the spike protein gene fragments of these SARS-associated coronavirus strains showed that the GZ01 isolate was phylogenetically distinct from other isolates, and compared with groups F1 and F2 of the T genotype, the isolates of BJ01 and CUHK-W1 were more closely related to the GZ01 isolate. It was interesting to find that two (A/G and C/T) of the five mutation loci occurred in the spike protein gene, which caused changes of Asp to Gly and Thr to Ile in the protein, respectively.

CONCLUSIONAttention should be paid to whether these genotype and mutation patterns are related to the virus's biological activities,epidemic characteristics and host clinical symptoms.

Objective:To evaluate the oncologic outcomes of different laparoscopic radical hysterectomy.Methods:From January 2011 to December 2014, the laparoscopic operation cases of cervical cancer at stage Ⅰb1, Ⅰb2, Ⅱa1 and Ⅱa2, including the histologic subtypes of squamous-cell carcinoma, adenocarcinoma and adenosquamous carcinoma, were collected in five clinical centers. The data were divided into two groups according to the surgical procedures, that is, modified laparoscopic-vaginal radical hysterectomy (mLVRH) and total laparoscopic radical hysterectomy (TLRH). The overall survival rate (OS), disease-free survival rate (DFS) at 5 years were retrospectively analyzed in this study.Results:There were 674 cases in total, including 377 cases of mLVRH, 297 cases of TLRH. (1) The OS at 5 years: the mLVRH was 96.1% and the TLRH was 92.0%, and the mLVRH was higher than that of TLRH （ P=0.010）. Stratify analysis， including stage of disease （Ⅰb1 and Ⅱa1）， histologic subtypes （squamous-cell carcinoma, adenocarcinoma）， lymph node metastasis， revealed that, ① Stage of disease: in stage Ⅰb1, the OS at five years of mLVRH was higher than that in TLRH group （98.6% vs 93.6%, P=0.012）. In stage Ⅱa1, there was significant difference between the two groups, the OS at five years of mLVRH and TLRH were 93.6% and 77.6% （ P=0.007）. ② Histologic subtypes: for the OS at five years of squamous-cell carcinoma, mLVRH and TLRH were 96.1% and 92.3%, and there was significant difference （ P=0.046）； for adenocarcinoma, the OS at five years were 91.0% and 88.6%， and there was no difference between two groups （ P=0.230）. ③ Lymph node metastasis： the mLVRH and TLRH with lymph node metastasis, the OS at five years were 98.6% and 96.4%; the mLVRH and TLRH without lymph node metastasis, the OS at five years were 89.3% and 80.8%. There were no significant differences between the two groups,respectively ( P=0.156， P=0.093）. (2) The DFS at 5 years: there was no significant difference between mLVRH and TLRH (94.1% vs 90.9%, P=0.220). Stratify analysis for stage of disease, the mLVRH group was higher than that in the TLRH group in stage Ⅰb1 (97.0% vs 92.8%, P=0.039). However, for stage Ⅱa1, there was no significant difference between mLVRH and TLRH group （88.2% vs 75.8%, P=0.074）. Conclusions:The results of this retrospective study indicated that different laparoscopy surgical procedures had diverse oncologic outcomes. The OS at 5 years of the mLVRH is superior to the TLRH. The DFS at 5 years in Ⅰb1 stage, the mLVRH is higher than the TLRH. Therefore, the modified laparoscopy is still an alternative surgery for early cervical cancer patients when following the principle of no-tumor-exposure.