Implementation of ovarian tissue cryopreservation in Hong Kong.

INTRODUCTION: Worldwide, >130 babies have been born from ovarian tissue cryopreservation (OTC) and ovarian tissue transplantation (OTT). Ovarian tissue cryopreservation can improve quality of life among young female cancer survivors. Here, we assessed the feasibility of OTC and subsequent OTT in Hong Kong via xenografts in nude mice. METHODS: This pilot study was conducted in a university-affiliated tertiary hospital. Fifty-two ovarian tissues were collected from 12 patients aged 29 to 41 years during ovarian surgery, then engrafted into 34 nude mice. The efficacies of slow freezing and vitrification were directly compared. In Phase I, non-ovariectomised nude mice underwent ovarian tissue engraftment. In Phase II, ovariectomised nude mice underwent ovarian tissue engraftment, followed by gonadotrophin administration to promote folliculogenesis. Ovarian tissue viability was assessed by gross anatomical, histological, and immunohistochemical examinations before and after OTC. Follicular density and morphological integrity were also assessed. RESULTS: After OTC and OTT, grafted ovarian tissues remained viable in nude mice. Primordial follicles were observed in thawed and grafted ovarian tissues, indicating that the cryopreservation and transplantation protocols were both effective. The results were unaffected by gonadotrophin stimulation. CONCLUSION: This study demonstrated the feasibility of OTC in Hong Kong as well as primordial follicle viability after OTC and OTT in nude mice. Ovarian tissue cryopreservation is ideal for patients who cannot undergo the ovarian stimulation necessary for oocyte or embryo freezing as well as prepubertal girls (all ineligible for oocyte freezing). Our findings support the clinical implementation of OTC and subsequent OTT in Hong Kong.

[Mechanism of Mycobacterium tuberculosis on interleukin-6 receptor 3'-untranslated region methylation in CD4+T cells].

Objective: To investigate the role and mechanism of DNA methylation in Mycobacterium tuberculosis (MTB lysate) -induced downregulation of interleukin-6 receptor(IL-6R) expression in CD4+T cells. Methods: A prospective study was conducted. Bisulfite sequencing (BSP) was applied to determine the methylation levels of CpG island in IL-6R promoter region and 3'untranslated region (3'UTR) region in CD4+T cells from peripheral blood mononuclear cells (PBMC) of control group (healthy person, n=10) and TB group (tuberculosis patients, n=10) in Shenzhen Third People's Hospital between 2019 and 2020. Quantitative reverse transcription-PCR (RT-qPCR) and Western blotting were used to detect the expression of IL-6R, DNMT1, DNMT3A and DNMT3B in MTB lysate-stimulated CD4+T cells and Jurkat E6-1 cells. Furthermore, PBMC in control group and Jurkat E6-1 cells activated by anti-CD3/CD28 antibody were stimulated by MTB lysates to detect the methylation levels of CpG island and IL-6R and DNMT expression. Transcriptional activity of differently methylation regions of IL-6R 3'UTR was detected by using luciferase reporter gene system. Results: IL-6R expression in TB group was lower than that in control group, but DNMT1 and DNMT3B expressions were higher than those in control group in CD4+T cells isolated from PBMC. There was no significant difference in the methylation rate of IL-6R promoter CpG island of CD4+T cells between control and TB group. However, the methylation rates of CpG island in 3'UTR region were significantly higher (P<0.001) in TB (69.5%±3.4%), compared with control (54.3%±4.7%). Besides, IL-6R expression was lower than unstimulated, while DNMT1 and DNMT3B expression was higher than unstimulated after MTB lysate-stimulation of activated control PBMC in vitro. The methylation rate of CpG island in IL-6R 3'UTR region of CD4+T cells increased from 58.8%±11.6% to 79.4%±10.9% (P<0.001) after MTB lysate-stimulated PBMC of the control. The same results were observed in the MTB lysate-stimulated CD4+T cells isolated from PBMC in control and Jurkat E6-1 cell line. Furthermore, IL-6R expression after co-treatment of the DNA methyltransferase inhibitor decitabine (5-aza) with MTB lysate was higher than that stimulated by MTB lysate alone. In addition, the methylation levels of CpG islands in the 3' UTR region of IL-6R were lower than those stimulated by MTB lysates alone after co-treatment of the DNA methyltransferase inhibitor decitabine (5-aza) with MTB lysates. The transcriptional activity of the fully unmethylated IL-6R 3'UTR CpG island reporter gene was higher than that of the fully methylated IL-6R 3'UTR CpG island. Conclusions: MTB lysates stimulation inhibited IL-6R expression transcriptionalely as well as on the protein level by inducing hypermethylation of CpG island in IL-6R 3'UTR region of CD4+T cells. The hypermethylation of CpG island in IL-6R 3'UTR region of CD4+T cells induced by MTB may be related to the increased expression of DNMT1 and DNMT3B.

[Prognostic evaluation value of (18)F-FDG PET-CT in Hodgkin's lymphoma after treatment].

Objective: To investigate the prognostic value of (18)F-fluorodeoxygen-D-glucose-positron emission tomography /computerized tomography ((18)F-FDG-PET-CT) in Hodgkin's lymphoma (HL) at the end of first-line treatment (PET-end), by comparing the ratio of maximum standardized uptake value (SUV(max)) of lesion and liver SUV (rLL), SUV(max) reduction between baseline PET (PET-0) and PET-end (ΔSUV(max)), and Deauville 5-point scale (5-PS). Methods: Patients with HL newly treated in our hospital from August 2006 to December 2015 were retrospectively analyzed. All the patients enrolled in the study underwent post-treatment FDG PET-CT. The rLL and ΔSUV(max) were calculated, and all the cases were scored using Deauville 5-PS. The receiver operating characteristic (ROC) approach was applied to identify the optimal cut-point value, and survival curves according to different PET-CT assessment methods were estimated using the Kaplan-Meier analysis. The prognostic efficacy of different PET-CT assessment methods was compared, and DeLong test was used to verify it. Kaplan-Meier method and multivariate analysis using the Cox proportional hazards model were performed to analyze the potential independent risk factors. Results: There were 5 patients progressed within a 3-year follow-up. In the three PET-CT assessment methods, the predictive value of rLL and Deauville 5-PS were significant effective. ROC analysis for rLL as a progression predictor showed an optimal cut-point of 1.29. Deauville 5-PS=4 and rLL=1.29 showed the best prognostic accuracy. The sensitivity of rLL and Deauville 5-PS were both 80.0%, and the specificity of each was 98.0% and 93.7%, respectively. The positive predictive value (PPV) and negative predictive value (NPV) of rLL were 66.7% and 98.7%, while the PPV and NPV of 5-PS were 44.4% and 98.7%. The 3-years progression-free survival (PFS) rates of rLL≥1.29 group and rLL<1.29 group were 33.3% and 98.7%, with significant difference (P<0.001). The 3-years PFS rates of post-treatment Deauville 5-PS<4 group and Deauville 5-PS≥4 group were 98.7% and 55.6%, with significant difference (P<0.001). The prognostic evaluation efficacy of rLL was positively correlated with that of Deauville 5-PS (r=0.75, P<0.05). Area under curves (AUC) of rLL and Deauville 5-PS were 0.93 (95%CI: 0.825-1.000) and 0.91 (95%CI: 0.757-1.000), respectively. DeLong test showed the significant difference between the two methods (P<0.05). The univariate analysis results showed that clinical baseline stage, post-treatment rLL and Deauville 5-PS were associated with the prognoses of HL patients (P<0.05). The multivariate analysis results showed that post-treatment rLL and Deauville 5-PS were independent prognostic factors of HL (P<0.05). Conclusions: The rLL and Deauville 5-PS are potential prognostic factors for HL response assessment. The new semi-quantitative method rLL has methodological advantages over visual analysis, and it is a good supplement for Deauville 5-PS. rLL can improve prognostic evaluation accuracy of PET-CT and is useful to early identify patients with HL at a high risk of relapsing after first-line treatment.

[Clinical efficacy of stent-assisted coil embolization for recurrent intracranial bifurcation aneurysms].

Objective: To evaluate the safety and efficacy of stent-assisted coil embolization in patients with recurrent intracranial bifurcation aneurysms,after initial simple coiling or microsurgical clipping. Methods: Clinical data of 20 patients with recurrent intracranial bifurcation aneurysms who initially underwent simple coiling or surgical clipping and subsequently re-treated by stent-assisted coiling embolization at the Radiology Intervention Department of Huashan Hospital between March 2009 and November 2019 were collected and analyzed retrospectively.There were 9 males and 11 females,with a median age of 55.5 years (range:33 to 71 years),including 17 aneurysms initially treated with simple coiling and 3 treated with surgical clipping.All cases were re-treated with stent-assisted coiling,15 using a single stent and 5 employing two stents in a Y-configuration.Peri-and post-operative complications and outcomes were evaluated.Mann-Whitney U tests were performed to compare the follow-up duration between initial treatment and re-treatment.Student's t tests were used to compare the parent artery angles before re-treatment, after re-treatment and at the last follow-up. The parent artery angle was defined using the proximal main trunk and the stented branch. Results: Immediate complete occlusion (Raymond Ⅰ) was achieved in 18 aneurysms (90.0%) while 2 aneurysms (10.0%) had a residual neck (Raymond Ⅱ).The median follow-up time(M(QR)) was 8.5(16.3)months,which had no significantly different from the initial treatment follow-up duration (15.5(27.0)months)(U=157.7,P=0.25). During the follow-up period,2 aneurysms (10.0%) with immediate post-operative residual necks recanalized again,including 1 aneurysm re-treated with the Y-configuration stent.Symptomatic thromboembolic complications occurred in 6 patients,including 4 re-treated with the Y-configuration stent.No peri-operative hemorrhagic complications occurred,along with no operation-related permanent disability or death. The parent artery angle increased significantly from pre-operative(90.1±21.1)°to post-operative and the last follow-up ((115.4±28.9)° and (132.6±26.8)°);t=5.14,P<0.01;t=7.78,P<0.01). Conclusion: For recurrent intracranial bifurcation aneurysms after initial surgical clipping or simple coiling,stent assisted coil embolization is proved to be safe and can decrease recurrence rate.

[Differences in efficacy between drug-eluting beads transbronchial arterial chemoembolization combined with systemic chemotherapy and systemic chemotherapy alone for unresectable lung squamous cell carcinoma].

Objective: To investigate the differences in efficacy between drug-eluting beads transbronchial arterial chemoembolization (DEB-BACE) combined with systemic chemotherapy and systemic chemotherapy alone for unresectable lung squamous cell carcinoma. Methods: Totally 60 cases of unresectable lung squamous cell carcinoma undergoing systemic chemotherapy in Yancheng Third People Hospital were retrospectively selected as the research object. According to patients' wishes, they were divided into chemotherapy-only group (group A) and combined treatment group (group B). Group A received gemcitabine combined with cisplatin chemotherapy. DEB-BACE was applied in the first half, and systemic chemotherapy was administered in the second half (starting 3 d after BACE). The first half and the second half of the chemotherapy dose were 1/2 of the drug dose in the chemotherapy alone group. The short-term efficacy, incidence of toxic side effects, peripheral blood T lymphocyte subsets, serum vascular endothelial growth factor (VEGF) levels, and survival time were compared between the two groups. Results: After 2 cycles of treatment, the total effective rates of group A and group B were 50.0% (15/30) and 76.7% (23/30) (P<0.05), the incidence of nausea and vomiting (63.3% vs 20.0%), decreased appetite (76.7% vs 43.3%), hair loss (86.7% vs 40.0%), and bone marrow suppression (40.0% vs 10.0%) in group A were higher than in group B (all P<0.05). After 2 cycles of treatment, the levels of CD3(+), CD4(+)and CD4(+)/CD8(+)in the two groups were higher than before treatment (group A: 47.7%±6.6% vs 52.3%±7.7%, 31.5%±4.9% vs 34.7%±5.8%, 1.05±0.24 vs 1.18±0.32; group B: 49.2%±7.0% vs 62.0%±14.0%,29.2%±5.5% vs 42.2%±7.3%, 1.07±0.26 vs 1.39±0.42; all P<0.05), while the level of CD8(+)was lower than before treatment (group A: 30.4%±5.4% vs 24.5%±4.8%; group B: 29.5%±4.1% vs 21.1%±4.5%; all P<0.05). The CD3(+), CD4(+), and CD4(+)/CD8(+) levels in group A were lower than those in group B (P<0.05), while CD8(+)level was higher than in group B (P<0.05). After 2 cycles of treatment, the VEGF levels in the two groups were lower than before treatment (group A: (423±85) vs (352±64) ng/L; group B: (404±114) vs (296±66) ng/L; P<0.05), and the VEGF level in group A was higher than that in group B (P<0.05). The 1-year survival rates of groups A and B were 54.9% and 77.9%, and the 2-year survival rates were 17.2% and 41.7% (Log rank χ(2)=4.750, P=0.029). Conclusion: DEB-BACE combined with systemic chemotherapy is superior to systemic chemotherapy in the treatment of unresectable lung squamous cell carcinoma. It can reduce toxic and side effects, improve immune function and prolong survival time, which is worthy of clinical application.

[An epidemiological survey on clinical features, self-management and cognitive level of elderly asthmatics in China].

Objective: To investigate the prevalence of asthma among the elderly people in China and to analyze the clinical features, self-management and cognitive level of elderly asthma patients. Methods: According to the multi-stage random cluster sampling methods, a total of 164 215 subjects were visited by a questionnaire in the last epidemiology survey from eight provinces (Beijing, Shanghai, Guangdong, Liaoning, Henan, Shanxi, Jiangsu, Sichuan provinces) and seven regions (north, northeast, southern china, east, south, southwest and northwest) in China from February 2010 to August 2012. 2 034 were diagnosed as asthma. The elderly patients aged ≥65 years were selected from the 2 034 asthma patients. The clinical characteristics, comorbidities, the status of asthma control and self-management and insights of the disease in elderly asthma patients were analyzed. Results: Among the 2 034 asthma patients, 584 (28.7%) were elderly asthmatics aged ≥65 years old and 1 450 (71.3%) were<65 years old. In the elderly asthma group, Early-onset asthma accounted for 439 (75.2%) and 145 (24.8%) were late-onset. The common clinical manifestations of elderly asthma patients were: chest distress 395 (67.6%), wheezing 304 (52.1%), cough 298 (51.0%). Common comorbidities of elderly asthmatics were: chronic obstructive pulmonary disease 144 (24.7%), allergic rhinitis 122(20.9%), gastroesopheal reflux disease (GERD) 114(19.5%), allergic conjunctivitis 86 (14.7%), eczema 82 (14.0%), chronic bronchitis 76 (13.0%). The Asthma Control Test (ACT) scores of elderly asthmatics and non-elderly asthmatics were (18.5±3.2) and (21.7±3.4) respectively. There was a significant difference between the two groups (P=0.042). Of the elderly asthmatics, only 13 (2.2%) patients monitored daily using a peak flow meter. 93 (15.9%) patients aware that asthma was characterized by chronic airway inflammation. 64 (11.0%) asthmatics understood that the treatment goal. Conclusions: The clinical manifestations of elderly asthmatics are atypical, especially paroxysmal wheezing. Asthma in elderly people causes more comorbidities and mortality. The self-management and cognitive level of patients with asthma needs to be improved.

[A survey on clinical characteristics and risk factors of severe asthma in China].

Objective: To investigate the clinical characteristics and risk factors of severe bronchial asthma in Chinese people over 14 years old. Methods: According to the multi-stage random cluster sampling methods, a total of 164 215 subjects were visited by a questionnaire in the epidemiology survey from eight provinces (Beijing, Shanghai, Guangdong, Liaoning, Henan, Shanxi, Jiangsu, Sichuan provinces) located in seven regions (north, northeast, east, central China, south, southwest and northwest) of China from February 2010 to August 2012. A total of 2 034 were diagnosed as asthma. The clinical characteristics and related risk factors of patients with severe asthma in China were analyzed. Results: Among all asthma patients, 560 were newly diagnosed, accounting for 27.5% (560/2 034) and the percentage of previously confirmed patients was 72.5% (1 474/2 034). A total of 145 were eligible for severe asthma, accounting for 9.8% (145/1 474) of previously confirmed asthmatics and 7.1% (145/2 034) of all asthmatics. 83.5% (121/145) severe asthmatics had at least one trigger factor. Correlation analysis showed that the risk factors of severe asthma were: smoking (OR=1.543, 95%CI: 1.250-1.814), obesity (OR=2.186, 95%CI: 1.972-2.354), petting (OR=2.135, 95%CI: 1.904-2.283), combined with allergic rhinitis (OR=3.456, 95%CI: 2.721-4.326), gastroesophageal reflux disease (OR=1.842, 95%CI: 1.682-2.140), bronchiectasis (OR=1.665, 95%CI: 1.347-1.912) or chronic obstructive pulmonary disease (OR=1.312, 95%CI: 1.171-1.694). Conclusions: The most common comorbidities in severe asthmatics in China are allergic rhinitis and gastroesophageal reflux disease. The risk factors of severe asthma include obesity, allergic rhinitis, gastroesophageal reflux disease, chronic obstructive pulmonary disease, bronchiectasis, smoking and petting.

[Clinicopathological characteristics of HPV(+) oropharyngeal squamous cell carcinoma].

Objective: To observe the clinicopathologic features of oropharyngeal squamous cell carcinoma associated with human papilloma virus (OPSCC-HPV) and discuss the role and value of different in situ hybridization (ISH) detection methods for HPV in pathologic diagnosis. Methods: Fifteen cases of OPSCC-HPV were collected from Department of Pathology, Beijing Tongren Hospital, Capital Medical University from January 2016 to August 2018. These cases were diagnosed in accordance with the WHO classification of head and neck tumors. The histopathologic features and the clinicopathologic data were retrospectively analyzed. Immunohistochemistry (two-step EnVision method) was done to evaluate the expression of p16, Ki-67 and p53. ISH was used to detect HPV DNA (6/11 and 16/18). RNAscope technology was used to evaluate the presence of HPV mRNAs (16 and 18). Results: The mean age for the 15 patients (8 males, 7 females) was 47 years (range from 30 to 69 years). OPSCC-HPV typically presentedat an advanced clinical stage, six patients had cervical lymphadenopathy (large and cystic), seven had tonsillar swelling, one had tumor at base of tongue, and one had odynophagia. Microscopically the tumors exhibited distinctive non-keratinizing squamous cell carcinoma morphology. Cervical nodal metastases were large and cystic, with thickening of lymph node capsules. OPSCC-HPV raised from crypt epithelium and extended beneath the tonsillar surface epithelial lining as nests and lobules, often with central necrosis. Tumor cells displayed a high N: C ratio, and high mitotic and apoptotic rates. Tumor nests are often embedded within lymphoid stroma, and may be infiltrated by lymphoid cells.Fifteen cases (15/15) were strongly positive for p16; Ki-67 index were 60%-90%; they were focally positive or negative for p53. Ten cases (10/10) were negative for HPV 6/11 DNA, and one case(1/10) was focally positive for HPV16/18 DNA. Eleven cases (11/11) were strongly positive for HPV16 mRNA, one case was focally positive for HPV18 mRNA. Conclusions: OPSCC-HPV is a pathologically and clinically distinct form of head and neck squamous cell carcinoma. OPSCC-HPV is associated with high-risk HPV (type 16) in all cases. Detection of high-risk HPV16 mRNA by RNAscope is of great significance in the final diagnosis and pathogen identification.

[The mobilization effect of IL-25 on eosinophil progenitor cells in allergic asthma].

Objective: Eosinophil progenitor cells (EoP) play a critical role in allergic airway inflammation in asthma. Previous studies have revealed that the expression of IL-25 receptor subunits (IL-17RA and IL-17RB) are increased on eosinophils (Eos) from allergic asthmatics upon allergen inhalation but few study has explored the role of IL-25 on EoP. Thus, in this research we examined the possible role of IL-25 on EoP in allergic asthmatics challenged by allergen, as well as in animal models where we verified the changes of newly produced Eos after IL-25 knockout. Methods: Asthmatics (n=14, during 2017-2018) who developed allergen-induced early and late responses were enrolled in this study. Blood was collected at pre-and 24 h post-challenge. Surface expression of IL-17RA and IL-17RB were evaluated by flow cytometry on EoP. In vitro migration assay was used to examine migrational responses of EoP and hematopoietic cells (HPC) from these subjects. In animal models, mice were grouped according to whether IL-25 was knock-out and whether mice were sensitized and challenged by ovalbumin (OVA) into asthmatic, control, knockout asthmatic and knockout control groups. Lung tissues, bronchoalveolar lavage flow (BALF) and bone marrow tissues of these mice were collected in order to evaluate airway inflammation and amount of newly produced (Brdu positive) and mature Eos. Results: EoP expressing IL-17RB were significantly increased after allergen inhalation in allergic asthmatics [(514±138) vs. (1146±450)/10(6) cells, pre-and post-challenge, F=6.819, P=0.022]. Pre-exposure to IL-25 primed the migrational responsiveness of EoP to stromal cell-derived factor (SDF)1α [(39.0±10.1)% vs. (73.0±7.9)%, control and IL-25 exposure groups respectively, P=0.021, 95% CI 5.19%~58.45%]. In OVA sensitized mice, knockout of IL-25 significantly reduced Eos and newly produced Eos percentage in the BALF [Eos, (7.8±2.0)% vs. (3.1±0.6)%, asthmatic and knockout asthmatic group respectively, P=0.002, 95% CI-7.57% to -1.98%; Brdu positive Eos, (50.0±7.6)% vs. (8.6±4.3)%, asthmatic and knockout asthmatic group respectively, P=0.011, 95% CI-72.41% to -10.27%], and newly produced Eos were also reduced in the bone marrow [(70.8±6.1)% vs. (1.3±1.3)%, asthmatic and knockout asthmatic group respectively, P=0.000, 95% CI -94.88% to -44.18%]. Conclusion: These results suggest an important role of IL-25 in allergen-induced EoP migration, local differentiation and eosinophilia in the airways.

Application of Multiple Genetic Markers in Determination of Full and Half Sibling Relationship： A Case Report.

ABSTRACT: Objective To investigate the application of the comprehensive use of multiple genetic markers in full and half sibling relationship testing through the identification of a case of suspected sibling relationship. Methods Genomic DNA were extracted from bloodstain samples from 4 subjects （ZHANG-1, ZHANG-2, male; ZHANG-3, ZHANG-4, female）. Autosomal STR loci, X-STR, Y-STR loci and polymorphisms of mtDNA HV-Ⅰ and Ⅱwere genotyped by EX20 STR kit, X19 kit, Data Y24 STR kit, and Sanger sequencing, respectively. Results According to autosomal STR based IBS scoring results, full sibling relationships were indicated among ZHANG-2, ZHANG-3 and ZHANG-4, but those were not indicated between ZHANG-1 and ZHANG-2 or ZHANG-3 or ZHANG-4. According to autosomal STR based FSI and HSI, with ITO method and discriminant function method, full sibling relationships among ZHANG-2, ZHANG-3 and ZHANG-4 were indicated, and half sibling relationships between ZHANG-1 and ZHANG-2 or ZHANG-3 or ZHANG-4 were also indicated. X-STR and mtDNA sequencing results showed that all the 4 samples came from a same maternal line, and Y-STR results showed that ZHANG-1 and ZHANG-2 did not come from a same paternal line, which supported the half sibling relationship between ZHANG-1 and ZHANG-2 or ZHANG-3 or ZHANG-4, verified by parental genotype reconstruction based on autosomal STR genotyping. Conclusion For the identification of sibling relationships, it is effective to have reliable results with the mutual verification and support of multiple genetic markers （autosomal STR, sex chromosomal STR and mtDNA sequence） and calculations （IBS, ITO, discriminant function method and family reconstruction）.

Health-related quality of life and its associated factors in Chinese middle-aged women.

OBJECTIVE: The aim of this study was to compare health-related quality of life (HRQOL) by menopausal stage and investigate its associated factors in middle-aged Chinese women. METHOD: This was a cross-sectional, community-based study involving 868 participants aged 40-60 years in Gongshu District, Hangzhou, Zhejiang, PR China. HRQOL was assessed by the Short-Form Health Survey (SF-36). The menopausal symptoms and sociodemographic characteristics were surveyed. RESULTS: The median (25-75th percentile) age of all participants was 51.24 (46.37-55.55) years. Differences were seen in domains of physical functioning, role-physical, bodily pain, general health and health transition by menopausal stage. The multivariate logistic regressions showed that there were associations between menopausal stage and HRQOL. Compared to premenopausal women, perimenopausal women had increased risks of having impaired functions in role-physical and health transition, and postmenopausal women were more likely to have impaired functions in physical functioning and health transition (p < 0.05 for all). Menopausal symptoms were negatively associated with HRQOL. Being married or co-habiting tended to relate to better general health. Being unemployed or retired tended to be associated with impaired role-physical. CONCLUSIONS: There was a difference in HRQOL by menopausal stage in middle-aged Chinese women. Menopause might exert a negative impact on HRQOL, adjusting for menopausal symptoms and sociodemographic factors.

[Value of PET/CT in the prognosis of extranodal NK/T cell lymphoma].

Objective: To explore the value of Positron-Emission Tomography/Computed Tomography (PET/CT) in the prognosis of extranodal NK/T cell lymphoma. Methods: The patients of NK/T cell lymphoma diagnosed from January 2007 to July 2016 in Department of Pathology of Beijing Tongren Hospital were enrolled in this study. Seventy-two in-hospital patients were examined on the invasion of adjecent tissue or organ by PET/CT. The PET/CT results were analyzed retrospectively. Kaplan-Meier method was used to analyze the prognostic value of the positive results by PET/CT on overall survival (OS). Results: There were 54 males and 18 females with median age of 44.5 years (13-75 years). According to Ann Arbor staging system, there were 16 cases (22.2%) in stage Ⅰ, 29 cases (40.3%) in stage Ⅱ, 6 cases (8.3%) in stage Ⅲ and 21 cases (29.2%) in stage Ⅳ. According to the IPI scoring system, there were 34 cases (47.2%) in the low risk group (0-1 point), 21 cases (29.2%) in the low-middle risk group (2 points), 16 cases (22.2%) in the middle-high risk group (3 points), and 1 case (1.4%) in the high risk group (4-5 points) . The median follow-up time was 29.2 months (1-118 months). The disease occured in unilateral nasal cavity in 26 cases (36.1%), bilateral nasal cavities in 36 cases (50.0%), nasopharynx, oropharynx and pharynx in 10 cases (13.9%). The tumors of 51 cases involved the surrounding tissue, including nasal wings in 29 cases (40.3%), nasal sinus in 29 cases (40.3%), maxillofacial soft tissue in 18 cases (25.0%), orbital in 12 cases (16.7%), maxilla and skull base in 8 cases (11.1%), eyelid in 6 cases (8.3%), brain tissue in 3 cases (4.2%), eyeball in 2 cases (2.8%). In addition, cervical and inguinal lymphadenopathy were found in 54 cases (75.0%) . Splenomegaly and hepatomegaly were found in 10 cases (13.9%) and 2 cases (2.8%), respectively. Survival analysis showed that the clinical stage and IPI were significantly associated the clinical prognosis (P=0.000, 0.001, respectively). Involvement of the maxillofacial soft tissue, eyelid, orbital, maxilla and skull base and brain tissue were significantly related to reduced the overall survival time (P=0.006, 0.000, 0.024, 0.001 and 0.015, respectively). Involvement of palate or tonsil, the nosewingand nasal sinus did not show significant shorter overall survival (P=0.091, 0.063, and 0.139, respectively). Cox regression multivariate analysis showed maxilla and skull base involvement was independent adverse prognostic factor (P=0.047). Conclusions: The PET/CT examination can accurately detect the extent of adjacent and distant tissues of tumor involvement of NK/T cell lymphoma by showing the tumor structure and metabolic status, thus has important value in the clinical staging and prognosis predication.

[Association of CYP19A1 gene rs7176005 single nucleotide polymorphism with breast cancer risk and clinicopathologic features of tumor].

Objective: The aim of this study was to investigate the association of the CYP19A1 rs7176005 single nucleotide polymorphism (SNP) with breast cancer risk and with clinicopathologic features of tumors. Methods: This study was conducted by including 138 patients with breast cancer (cancer group), those who diagnosed as primary breast cancer after operation by pathology. There were 293 cases in the group of benign breast disease which was presented as a solid mass by the color ultrasound and pathologically diagnosed as "fibroadenoma or adenosis" (benign breast disease group), the cases were paired with breast cancer patients by age±5 in the same period, and there were 259 cases in the group of healthy control who received routine physical examination during the same period and were paired with breast cancer patients by age±5 without any detection of breast related diseases (healthy control group) at West China hospital between September 2012 and November 2016. The CYP19A1 rs7176005 SNP was detected by a direct sequencing method. Hardy-Weinberg test was used to analyze the genetic balance of the 3 groups. Chi square test was used to compare the distribution of rs7176005 genotypes between the 3 groups, and the differences of clinicopathological features in breast cancer patients carrying different genotypes. Results: The ages of the breast cancer cases, the benign breast disease group and the healthy control group were (44.69±8.09), (42.33±11.44) and (41.92±9.61) years old, respectively. Hardy-Weinberg equilibrium test identified that the composition ratios of alleles C and T in breast cancer group, benign breast disease group and healthy group were not statistically significant (χ(2) values were 0.83, 0.34 and 0.04, respectively, P values were 0.363, 0.561, and 0.852, respectively). All the three groups met the genetic balance, had consistency and could represent the population. Among the 138 cases of breast cancer, the CYP19A1 rs7176005 SNP was significantly associated with the diameter of the tumor (P=0.031). The majority of tumor size was <2 cm in patients who carrying TT and CT genotypes, and the proportion was 75% (12/16) and 58% (40/69), respectively. While those patients with TT genotype were mainly >2 cm and ≤5 cm, and the proportion was 51% (27/53). The distribution of TNM stage among patients with different genotypes was also statistically significant (χ(2)=11.19, P=0.025). The most common stage was Ⅱ in Patients who carrying CC and CT genotypes, and the proportion was 45.3% (24/53) and 52.2% (36/69), respectively. While those patients with TT genotype was mainly in stage Ⅰ and the proportion was 56.3% (9/16). Conclusion: Though the CYP19A1 rs7176005 SNP is not associated with breast cancer development, breast cancer patients with the C allele exhibit a high tumor growth rate and large diameters.

Immunotolerant indoleamine-2,3-dioxygenase is increased in condyloma acuminata.

BACKGROUND: The tryptophan-depleting enzyme indoleamine-2,3-dioxygenase (IDO) is critical for the regulation of immunotolerance and plays an important role in immune-associated skin diseases. OBJECTIVES: To analyse the level of IDO in condyloma acuminata (CA) and its role in this condition. METHODS: IDO expression was assessed in the skin and peripheral blood of healthy controls and patients with CA. To assess the role of skin IDO in immunity, the ability of isolated epidermal cells to metabolize tryptophan and the influence on polyclonal T-cell mitogen (PHA)-stimulated T-cell proliferation were explored. RESULTS: IDO median fluorescence intensities in peripheral blood mononuclear cells from patients with CA were similar to those from healthy controls. Immunohistochemistry showed that IDO+ cells were rare in normal skin and the control skin of patients with CA, but were greatly accumulated in wart tissue. Most fluorescence signals of IDO+ cells did not overlap with those of CD1a+ Langerhans cells. Human papillomavirus (HPV) DNA probe in situ hybridization showed a large number of IDO+ cells in the HPV- site. Keratinocytes in the skin of healthy controls and the circumcised skin of patients with CA could minimally transform tryptophan into kynurenine, but IDO-competent epidermal cells from warts could transform tryptophan. In addition, these IDO-competent epidermal cells could inhibit PHA-stimulated T-cell proliferation. The addition of an IDO inhibitor, 1-methyl-d-tryptophan, restored the inhibited T-cell proliferation. CONCLUSIONS: Abnormally localized high IDO expression might be involved in the formation of a local immunotolerant microenvironment.

Phenotypic classification of gastric signet ring cell carcinoma and its relationship with K-ras mutation.

We aimed to analyze gastric signet ring cell (SRC) carcinoma subtypes by investigating gastric and intestinal phenotypic marker expression, and explore the relationship between phenotype and K-ras mutation. Immunohistochemistry was performed on 163 SRC carcinoma patient specimens to detect gastric (MUC1, MUC5AC, and MUC6) and intestinal (MUC2 and CDX2) phenotypic markers, and tumors were classified into gastric (G), intestinal (I), and gastrointestinal (GI) phenotypes. DNA was extracted from the formalin-fixed, paraffin-embedded tumor samples, and K-ras mutations in codons 12, 13, and 61 were identified using polymerase chain reaction-based direct DNA sequencing. G, GI, and I phenotypes were observed in 63 (38.6%), 71 (43.5%), and 29 cases (17.8%), respectively. Expression of MUC2 was significantly associated with invasion depth and lymph node metastasis (P = 0.001 and 0.002, respectively), whereas that of CDX2 significantly corresponded to tumor size and submucosal invasion (P = 0.004 and 0.001, respectively). MUC5AC expression was inversely associated with gastric wall invasion (P = 0.001). Intestinal phenotypic marker expression was positively associated with gastric wall invasion and lymph node metastasis. K-ras mutations, all of which were in codon 12, were detected in 20 (12.27%) tumors, were significantly associated with the I phenotype, and exhibited an inverse relationship with MUC5AC and MUC6 expression. I-phenotype SRC carcinomas should be distinguished from those of the G phenotype because of their increased malignancy regarding invasion and metastasis, and higher K-ras aberration rate. The different K-ras mutation frequencies observed imply distinct genetic mechanisms in the carcinogenesis of I- and G-phenotype gastric SRC carcinomas.

[Inhibition of G9a attenuates cell proliferation via the mitochondrial apoptosis pathway in lung adenocarcinoma].

Objective: The aim of this study is to investigate the effect of G9a inhibitor BIX-01294 on attenuating cell proliferation in human lung adenocarcinoma A549 cell line and the underlying molecular mechanism. Methods: Treated with BIX-01294, the growth and proliferation of A549 cells were detected by MTT assay and colony formation assay, and its impact on cell apoptosis was analyzed using flow cytometry. By Western blot, we explored the alterations in the expression of apoptosis-related proteins and the G9a catalysate, H3K9me and H3K9me2. In addition, in the pretreatment with caspase inhibitor Z-VAD-FMK, we detected the apoptotic dependence of BIX-01294 attenuating impact on A549 cell proliferation. Results: Compared with the control group, the histone methyltransferase G9a inhibitor BIX-01294 attenuated cell proliferation in A549 cells in a dose- and time-dependent manner. There were 42.5±8.7 colonies after BIX-01294 (10 µmol/L) treatment for 7 days, while 172.7±23.0 colonies in the control group, with a statistical significance (P<0.05). After treatment with BIX-01294 (10 µmol/L) for 24 hours, the cell apoptotic rate was(47.6±8.4)%, with a significant difference in comparison with the control group [(7.2±3.6)%, P<0.05]. The expression of G9a catalysate, H3K9me and H3K9me2 was downregulated, the same with anti-apoptotic protein Bcl-2, while the proteins in mitochondrial apoptosis pathway, Bax, Bak and cleaved caspase-9, were upregulated, so was the expression of cleaved caspase-3 and cleaved PARP, and there was no alteration in the expression of cleaved caspase-8, which is a protein related with death receptor apoptosis pathway. Furthermore, after Z-VAD-FMK pretreatment, the cell apoptotic rate was decreased significantly, and the expression of apoptosis-related proteins were downregulated. Conclusions: Our results indicate that BIX-01294 can attenuate cell proliferation in lung adenocarcinoma, and it can be considered as one of the underlying mechanisms, the apoptosis may be induced by activating mitochondrial pathway.

[Chinese expert consensus for non-antiinfective effects and clinical use of macrolides].

Important/potential value of macrolides has been proved in the management of chronic respiratory diseases by increasing basic and clinical trials.Through three face-to-face discussions, 10 experts examined important data and drafted this consensus related to macrolides: (1) mechanism of non-antiinfective effects; (2) clinical use in chronic respiratory diseases; (3) cautions of long-term use.The mechanism out of non-antiinfective effects includes anti-inflammatory effect, modifying airway secretion, immune-regulation related to antibacterial effect, corticoid saving effect and anti-viral effect.The efficacy of long-term use of low-dose macrolides is definitely confirmed in diffuse panbronchiolitis, chronic rhinosinusitis. It is considerably used in bronchiectasia, cystic fibrosis, severe asthma and chronic obstructive pulmonary disease. Further studies should be conducted in cryptogenic organizing pneumonia and respiratory viral infection. It should be paid attention to its possible adverse effects (including drug interactions, cardiac toxicity, ototoxicity and disturbance of intestinal flora) and drug resistance in long-term use.A Chinese consensus for non-antiinfective effects and clinical use of macrolides is developed for the first time, which aims to expand their rational use and the further research.

[The relationship between smoking status and epidermiology of asthma in people aged over 14 years in China].

Objective: To study the relationship between bronchial asthma and smoking status in Chinese people. Methods: Asthma epidemiological survey and stratified-cluster-random method survey were performed in residents over 14 years in 8 provinces (cities) of China from February 2010 to August 2012. Asthma was diagnosed based upon case history, clinical signs and lung function test. Smoking status was investigated by questionnaire. Results: Sampling population was 180 099 and 164 215 were valid. A total of 2 034 subjects were diagnosed as asthma including 79 692 men and 84 523 women. The overall prevalence rate of asthma was 1.24% (2 034/164 215). Smokers were 23.8% (39 137/164 215) in the whole population. Smokers were 34.5% (702/2 034) in asthmatic patients, compared with 23.7% (38 435/162 181) in no-asthmatic population. The incidence of asthma was 1.79% and 1.06% in smokers and non-smokers respectively (P<0.001), suggesting that OR of smoking was 1.70 (95% CI 1.55-1.86, P<0.001). According to asthma control test (ACT) score, the level of asthma control in non smoking group was higher than that in smoking group(43.2% vs 35.3%). The times of hospitalization due to acute exacerbations(0.51 vs 0.41 events/person/year), total hospitalization rate(27.35% vs 20.12%), annual emergency room visits (0.80 vs 0.60 events/person/year) and emergency room visit rate (31.77% vs 24.47%) were all much higher in smoking asthmatic patients than those in non smoking asthmatic patients, indicating that the level of asthma control in smoking patients was significantly worse than in non smoking patients. Conclusions: The smoking rate in Chinese people over 14 years is still high. The prevalence rate of asthma in smokers is significantly higher than that of non-smokers. The level of asthma control in smokers is significantly worse than that in non smokers.