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 Isolating and purifying the major allergens from Metapenaeus ensis and identifying its immunoreactivity for diagnostic application. Methods The proteins extracted from Metapenaeus ensis were purified by DEAE-52 ion-exchange chromatography and Sephadex G-100 gel filtration. The eluted peaks were detected by dot-ELISA,and the immunoreactivity of the purified allergens was detected by ELISA with the sera from patients allergic to shrimp. The molecular weights of the purified allergic proteins were detected by SDS-PAGE. Results Two major allergic proteins which have highly immunoreactivity in ELISA detection were purified from Metapenaeus ensis. The molecular weights of which were 36 and 68kDa,respectively. Conclusion The major allergens of Metapenaeus ensis purified by ion-exchange chromatography and gel filtration have effective immunoreactivity and can be used as specific antigen in ELISA reagent.

33 cases were treated by spotted—moxibustionwith Zhuang medicated threads,the effective rate be-ing 84.6～85.0%.The longterm effect was also satis-factory.This therapy can also markedly raised the in-filtration pressure of nocturnal urination.There wasno obvious relation between the therapeutic effect andsyndrome typing.

Objective:To explore the application effect of self-management ability-oriented focus solution model in children with nephrotic syndrome, and to observe the optimal value of self-management and coping style for children with nephrotic syndrome.Methods:This was a class trial study. A total of 96 children with nephrotic syndrome in Children′s Hospital of Nanjing Medical Universityfrom March 2018 to April 2020 were selected as the research object by convenient sampling method, 48 cases of children from March 2018 to February 2019 for the control group, using conventional psychological counseling, 48 cases of children from March 2019 to April 2020 for the observation group, with self management ability oriented focus solution mode. The self-management and response changes after 2 months of intervention were compared between the two groups.Results:The scores of treatment management, psychological management, diet management and social activity inself-management behavior after intervention were (32.79 ± 2.94), (19.43 ± 1.76), (22.34 ± 1.64), (39.79 ± 2.97) points in the observation group, and (27.42 ± 3.25), (16.79 ± 1.52), (18.46 ± 1.58), (35.28 ± 2.64) points in the control group, the differences were statistically significant ( t values were 7.86-11.80, all P<0.05). The scores of escape, acceptance, negative emotional response, and total score after intervention were (12.41 ± 2.15), (20.41 ± 1.85), (25.89 ± 1.68), (99.37 ± 8.94) points in the observation group, and (11.22 ± 3.04), (19.30 ± 2.09), (24.80 ± 2.10), (95.12 ± 9.02) points in the control group, the differences were statistically significant ( t values were 2.21-2.81, all P<0.05). Conclusions:The self-management-oriented focus solution model for children with nephrotic syndrome can promote the improvement of self-management behavior, and promote the improvement of coping methods in terms of avoidance, acceptance and negative emotional response, which has reference and complementary value for the care of children with nephrotic syndrome.

Under the leadership and high-level promotion of the central and local governments, the construction of national regional medical centers(NRMCs) in China has achieved initial results, and has received recognition and high attention from all sectors of society. The current focus of NRMCs construction has progressed from the project selection to the evaluation of construction effectiveness and the establishment of corresponding operating mechanisms. On the basis of analyzing the characteristics of NRMCs construction and reviewing the evaluation of NRMCs construction at the national level, this paper systematically introduced the monitoring index system and its application of NRMCs construction in Beijing Children′s Hospital, and put forward corresponding policy suggestions. The monitoring of the construction indexes of NRMCs could provide a basis and support for hospitals to identify problems in time, seek government support, and carry out target management, and has reference for promoting the effective construction of NRMCs.

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.

Objective:Tumor-treating fields (TTFields) is a kind of non-invasive anti-mitotic tumor therapy, which has been approved for patients with newly diagnosed and recurrent glioblastoma. This study aims to explore the efficacy and safety of TTFields in high-grade gliomas in clinical practice settings.Methods:The clinical data of 15 patients with recurrent glioma and 9 patients with newly diagnosed high-grade glioma admitted to our center from April 2019 to January 2021 were retrospectively analyzed. All patients accepted TTFields≥1 month. Follow-up was performed for 5.3 months (ranged from 2.3 to 10.7 months); Response Assessment in Neuro-Oncology Working Group (RANO) criteria was used to evaluate the glioma responses. The progression-free survival (PFS) and overall survival (OS) were calculated according to Kaplan-Meier method. Common Terminology Criteria for Adverse Events v5.0 (CTCAE v5.0) and TTFields related skin adverse reaction (dAE) criteria were used to evaluate the adverse events. Quality of life questionnaire-core 30 (QLQ-C30) and QLQ-brain cancer module (QLQ-BN20) questionnaires were used to evaluate the health-related quality of life (HRQoL). Treatment compliance was evaluated by data on the use of NovoTTF-200A devices, and calculated as a percentage of daily TTFields usage.Results:The median duration of TTFields was 4.2 months (ranged from 1.0 to 10.7 months), with a median compliance rate of 91.5% (67.0%-97.0%). TTFields was used alone in 2 patients and used with combination of chemotherapy in 22 patients. From follow-up to April 2021, 14 patients had stable symptoms and 10 had disease progression (8 died). The median PFS and OS of recurrent patients were 5.9 months ( 95%CI: 3.3-8.6 months) and 8.5 months ( 95%CI: 3.2-13.8 months), respectively; and the median PFS and OS of newly diagnosed patients were both 10.7 months (without 95%CI). The common adverse events included grading 1 dAE (58.3%) and grading 2 dAE (12.5%), without grading 3 or 4 dAE, manifested as contact or allergic dermatitis, erosion, folliculitis and ulcers. And 87.5% patients had stable HRQoL. Conclusions:The preliminary results showed that the survival of recurrent high-grade glioma patients treated by TTFields is similar to that reported in foreign literature; and the newly diagnosed patients need further survival follow-up. The patients' treatment compliance and safety are good. The dAE incidence (grading 1-2) is higher than that reported in the literature, and the toxicity was acceptable.

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,ll (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 50-0 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.