Biomarkers for early screening and diagnosis of breast cancer: a review / 生物工程学报

The estimated new cases of breast cancer (BC) patients were 2.26 million in 2020, which accounted for 11.7% of all cancer patients, making it the most prevalent cancer worldwide. Early detection, diagnosis and treatment are crucial to reduce the mortality, and improve the prognosis of BC patients. Despite the widespread use of mammography screening as a tool for BC screening, the false positive, radiation, and overdiagnosis are still pressing issues that need to be addressed. Therefore, it is urgent to develop accessible, stable, and reliable biomarkers for non-invasive screening and diagnosis of BC. Recent studies indicated that the circulating tumor cell DNA (ctDNA), carcinoembryonic antigen (CEA), carbohydrate antigen 15-3 (CA15-3), extracellular vesicles (EV), circulating miRNAs and BRCA gene from blood, and the phospholipid, miRNAs, hypnone and hexadecane from urine, nipple aspirate fluid (NAF) and volatile organic compounds (VOCs) in exhaled gas were closely related to the early screening and diagnosis of BC. This review summarizes the advances of the above biomarkers in the early screening and diagnosis of BC.

Advances in predictive biomarkers associated with immune checkpoint inhibitors for tumor therapy / 生物工程学报

Malignant tumors are diseases that seriously threaten human health and social development. Traditional tumor therapies such as surgery, radiotherapy, chemotherapy and targeted therapy cannot fully meet the needs of clinical treatment, and emerging immunotherapy has become a research hotspot in the field of tumor treatment. Immune checkpoint inhibitors (ICIs) have been approved as a tumor immunotherapy method for the treatment of various tumors, such as lung cancer, liver cancer, stomach cancer and colorectal cancer, etc. However, during the clinical use of ICIs, only a small number of patients experienced durable responses, which also led to drug resistance and adverse reactions. Therefore, the identification and development of predictive biomarkers is crucial to improve the therapeutic efficacy of ICIs. The predictive biomarkers of tumor ICIs mainly include tumor biomarkers, tumor microenvironment biomarkers, circulation-related biomarkers, host environmental biomarkers and combinatorial biomarkers. They are of great significance for screening, individualized treatment and prognosis evaluation of tumor patients. This article reviews the advances of predictive markers for tumor ICIs therapy.

Analysis of pathogenic microorganisms in 64 077 cases of umbilical cord blood / 中国输血杂志

【Objective】 To investigate the detection of pathogenic microorganisms in umbilical cord blood and maternal blood from 2012 to 2021, so as to improve the collection of umbilical cord blood and guarantee the safety of umbilical cord blood hematopoietic stem cells (HSC) . 【Methods】 Detection results of pathogenic microorganisms of umbilical cord blood and maternal blood among 64 077 cases from Tianjin Cord Blood Bank from 2012 to 2021 were retrospectively analyzed. 【Results】 A total of, 2 072 cases (3.23%) were detected positive, among which, 184 cases (0.29%) were positive for aerobic bacteria culture, 1 504 cases (2.34%) were positive for anaerobic bacteria culture, and 384 cases (0.60%) were positive for both aerobic and anaerobic bacteria culture. From 2012 to 2021,the overall positive rate showed a downward trend, with a difference in the positive rate between each year (P<0.05). The positive rate of anaerobic bacteria was higher than that of aerobic bacteria and that of anaerobic and aerobic bacteria (P<0.05). After Gram staining, the microscopic detection rate of bacterial positive samples was highest in G- bacilli, followed by G+ bacilli, G+ cocci, G- cocci and others. Among the 64 077 cases, 169 cases (0.26%) showed reactivity in cord blood tests and 1 231 cases (1.92%) showed reactivity in maternal blood tests. Umbilical cord blood and maternal blood HIV-Ag/Ab tests showed reactivity after initial screening. After confirmation by Western blotting, there was 1 case of uncertain maternal blood, while the rest were negative. The reactive rates of anti-TP (0.12%) and anti- HCV (0.11%) in umbilical cord blood were higher than those of HBsAg (0.03%) and CMV-IgM (1/64 077).There was a difference in the reactive rate of anti-TP detection in umbilical cord blood between different years (P<0.05),while there was no statistically significant difference in that of HBsAg, anti-HCV and CMV-IgM (P> 0.05).The reactive rate of HBsAg in maternal blood (1.38%) was higher than that of CMV-IgM(0.29%) , anti-TP(0.13%) and anti-HCV (0.12%) . There were differences in the reactive rates of HBsAg, anti-HCV ,and anti-TP in maternal blood among different years (P<0.05),and that of HBsAg showed a decreasing trend, while the reactive rate of CMV-IgM was not statistically significant (P>0.05). The reactive rates of HBsAg and CMV-IgM detected in maternal blood were significantly higher than those in umbilical cord blood (P<0.05) . The reactive rates of anti-HCV and anti-TP in maternal blood were consistent with those in umbilical cord blood (P>0.05). 【Conclusion】 The reactive rates of anti-HIV and CMV-IgM in cord blood, and that of anti-HIV in maternal blood are low, but those of anti-TP and anti-HCV in cord blood are relatively high. The reactive rate of HBsAg is high in maternal blood,but with a downward trend,but low in umbilical cord blood due to maternal-infantile transmission blocking. The detection of transfusion transmitted pathogens and bacteria plays a critical role on the safety of umbilical cord blood HSCs. Effective detection of transfusion transmitted pathogens and culture of bacteria are the key to ensure the quality of umbilical cord blood, which can improve the safety of umbilical cord blood HSCs transplantation.

Added value of T 1-weighted StarVIBE sequence for PET/MR image quality / 中华核医学与分子影像杂志

Objective:To explore the added value of T 1-weighted stack-of-stars volumetric interpolated body examination (StarVIBE) sequence on PET/MR image quality. Methods:A retrospective analysis was performed on 60 patients (42 males, 18 females; age 11-86 (58±12) years) who underwent 18F-FDG PET/MR examination and with positive PET results in Ruijin Hospital, Shanghai Jiao Tong University School of Medicine from April 2020 to April 2021. All patients completed StarVIBE sequence collection, and volumetric interpolated body examination (VIBE) sequence was used as control. StarVIBE and VIBE sequence images were evaluated independently using five-point method by two physicians. The evaluation was carried out from six aspects: lesion display, lesion boundary display, vascular around lesions display, fusion level with PET image, image artifact and overall image quality. Wilcoxon signed rank test was used to compare the image quality of the two sequences, and Kappa test was performed to assess the consistency of the image quality scores between the two physicians. Results:There were 26 cases with cervical lesions, 14 cases with chest lesions, 7 cases with abdomen lesions and 13 cases with pelvic lesions. The scores of lesion display (4.0(3.8, 4.5) vs 3.5(3.0, 4.0)), lesion boundary display (4.0(4.0, 4.0) vs 3.0(3.0, 3.5)), vascular around lesions display (5.0(4.0, 5.0) vs 4.0(3.5, 4.5)), fusion level with PET image (5.0(5.0, 5.0) vs 4.5(4.0, 5.0)), image artifact (4.5(4.0, 5.0) vs 4.5(4.0, 5.0)) and overall image quality (5.0(4.0, 5.0) vs 4.0(4.0, 4.0)) of StarVIBE sequences were better than those of VIBE sequences ( z values: 3.77-6.54, all P<0.001). On the vascular around the lesions display, the scores of StarVIBE were significantly better than those of VIBE sequence in the neck (5.0(4.5, 5.0) vs 3.0(2.7, 3.5); z=4.49, P<0.001) and chest (4.5(4.3, 4.7) vs 4.0(3.6, 4.3); z=3.10, P=0.002). As for image quality, the scores of StarVIBE were also significantly better than those of VIBE in neck (5.0(4.5, 5.0) vs 4.0(3.7, 4.5); z=4.36, P<0.001) and chest (5.0(5.0, 5.0) vs 4.0(4.0, 4.5); z=3.02, P=0.003). In abdominal lesions, the score of StarVIBE was higher than that of VIBE in blood vessels (4.5(3.5, 5.0) vs 4.0(3.5, 4.5); z=2.07, P=0.038), and there was no difference between score of overall image quality (4.0(3.7, 4.5) vs 4.0(3.5, 4.5); z=0.27, P=0.785). The score of overall image quality of pelvic StarVIBE sequence was better than that of VIBE sequence (5.0(4.5, 5.0) vs 4.0(4.0, 4.5); z=2.12, P=0.034). Kappa value of image quality score between two physicians was 0.554, indicating moderate consistency. Conclusion:In whole-body PET/MR imaging, StarVIBE sequence can significantly improve the image quality of cervical, thoracic and pelvic lesions when comparing with VIBE sequence.

Circular RNA ame_circ_000115 regulates expression of genes in larval gusts of Apis mellifera ligustica stressed by Ascosphaera apis / 生物工程学报

Circular RNAs (circRNAs) are a new class of non-coding RNAs, which have been confirmed to regulate insect gene expression and immune response through multiple manners such as competing endogenous RNA (ceRNA) regulatory network. Currently, function of circRNA in honey bee immune response remains unclear. In this study, PCR and Sanger sequencing were performed to validate the back splicing (BS) site of ame_circ_000115 (in short ac115). RT-qPCR was used to detect the expression profile of ac115 in larval guts of Apis mellifera ligustica stressed by Ascosphaera apis. Dual-luciferase reporter gene assay was conducted to verify the binding relationship between ac115 and ame-miR-13b. Interference of ac115 in larval guts was carried out by feeding specific siRNA, followed by determination of the effect of ac115 interference on expression of six genes relevant to host immune response. The results confirmed the existence of BS site within ac115. Compared with the un-inoculated group, the expression of ac115 in 4-day-old larval gut of the A. apis-inoculated group was up-regulated with extreme significance (P < 0.000 1), while that in 5- and 6-day-old larval guts were significantly up-regulated (P < 0.05). The brightness of specific band for ac115 in 4-, 5- and 6-day-old larval guts of the siRNA-circ_000115-fed group gradually became weak, whereas that of the siRNA-scrambl-fed group was pretty high without obvious variation. Compared with that of the siRNA-scramble-fed group, the expression of ac115 in 4-day-old larval gut of the siRNA-circ_000115-fed group was significantly down-regulated (P < 0.05), whereas that of the 5- and 6-day-old larval guts were down-regulated with extreme significance (P < 0.001). Ame-miR-13b was truly existed and expressed in A. m. ligustica larval guts, and there was true binding relationship between ac115 and ame-miR-13b. Compared with that of the siRNA-scramble-fed group, the expression of antimicrobial peptide genes hymenoptaecin and abaecin in 6-day-old larval gut of the siRNA-circ_000115-fed group was significantly up-regulated (P < 0.05), while that of ecdysone receptor (Ecr) was down-regulated with extreme significance (P < 0.01). These results indicate that ac115 is truly expressed in A. m. ligustica larval guts, BS site truly exists within ac115, and effective interference of ac115 in A. m. ligustica larval guts can be achieved via feeding siRNA. Moreover, ac115 potentially regulates Ecr expression through adsorption of ame-miR-13b and expression of hymenoptaecin and abaecin using a non-ceRNA manner, further participating in host stress-response.

Study on the efficacy and safety of combination of sufficient argon plasma coagulation cauterization and clipping in the treatment of colonic diverticular bleeding / 中华消化杂志

Objective:To evaluate the efficacy and safety of combination of sufficient argon plasma coagulation(APC) cauterization and clipping in the treatment of colonic diverticular bleeding.Methods:From June 2018 to April 2022, the clinical data of patients were retrospectively analyzed, who visited Department of Gastroenterology of Air Force Medical Center due to overt gastrointestinal bleeding, and were confirmed or suspected to have colonic diverticular bleeding and received combination of sufficient APC cauterization and clipping treatment. The deadline for follow-up was September 30, 2022. During the follow-up after endoscopic treatment, the re-bleeding rate, hemoglobin level difference between the last follow-up and before treatment, wound healing under colonoscopy as well as the intraoperative and postoperative complications of patients were statistically analyzed. Descriptive analysis was used for statistical analysis.Results:A total of 15 patients were enrolled, including 13 males and 2 females, aged (60.8±14.8) years old. The course of the disease was 1 day to 13 years. A total of 145 colonic diverticula of 15 patients were treated under endoscopy. The median follow-up time was 14.5 months (5.3 to 49.5 months) months. Among the 15 patients, 12 patients received endoscopic therapy once and no bleeding occurred till the end of follow-up. Three patients suspected with diverticular bleeding received a second endoscopic treatment because of bleeding at the 12 days, 3 months and 8 months after the first treatment, respectively.No rebleeding occurred after the second endoscopic therapy till the end of follow-up. The re-bleeding rate of the first treatment was 3/15 and the re-bleeding rate of re-treatment was 0. At the end of follow-up, the hemoglobin concentration increased (35.9±26.3) g/L compared with that before the treatment. Two patients had perforation during operation and were closed with multiple titanium clips. There was no abdominal pain or other symptoms after operation. And the patients were discharged 3 and 4 days after treatment, respectively. Two patients suffered short-term postoperative wound bleeding and successful hemostasis was achieved after endoscopic treatment. One patient developed postoperative infection and the symptoms disappeared after anti-infection treatment.Conclusions:Combination of sufficient APC cauterization and titanium clipping is safe and effective in the treatment of colonic diverticular bleeding. For patients with dominant diverticular hemorrhage, or patients with recurrent gastrointestinal bleeding, if other etiology are excluded and colonic diverticular bleeding is highly suspected, the combination of sufficient APC cauterization and titanium clipping under endoscopy is feasible.

Data mining and analysis of febuxostat-induced cardiac ADR signals / 中国药房

OBJE CTIVE To mine and analyze t he cardiac adverse drug reaction （ADR）signals induced by febuxostat in post-marketing experience ，and to provide reference for rational drug use in clinic. METHODS Reporting odds ratio （ROR） method was used to mine the ADR signals induced by febuxostat from the FDA Adverse Event Reporting System during the first quarter of 2009 to the fourth quarter of 2020；the information of cardiac disease signals was counted and analyzed. RESULTS A total of 209 ADR signals were detected in 8 282 adverse drug event （ADE）reports with febuxostat as the primary suspected drug ， involving 27 cardiac signals and 754 ADE reports. The most reported signals were symptoms （262 reports），including dizziness ， oedema peripheral，chest pain ，palpitations and gravitational oedema and so on ，followed by coronary atherosclerotic heart disease signal，heart failure signal ，arrhythmia signal ，sudden cardiac death signal （233，157，90，12 reports，respectively）. More than half of the signals were mentioned in the drug instructions ，while the unmentioned signals were mainly kinds of cardiac failure ， arrhythmia and extrasystoles ，etc. The patients with cardiac ADEs who received febuxostat were more male than female ，and the age was 60 and over ；the drug dosage was mostly 40 mg/d or 80 mg/d as recommended in the drug instructions ，and cardiac ADEs mostly occurred within 1 month of medication. CONCLUSIONS Routine attention should be paid to the cardiac safety of febuxostat during medication ，further evaluation and validation of febuxostat-induced cardiac ADR signals are still needed.

Therapeutic effects of thoracoscopic ultrasound-guided pigtail drainage for tuberculous pleural effusion in elderly patients / 中华老年医学杂志

Objective:To investigate the therapeutic effects of ultrasound-guided pigtail drainage under thoracoscopy in elderly patients with tuberculous pleural effusion.Methods:Elderly patients with tuberculous pleural effusion treated at Zhengzhou Sixth People's Hospital between April 2018 and May 2020 were selected.With a method using odd and even numbers, 100 elderly patients with tuberculous pleural effusion were randomly divided into a study group(odd numbers)and a conventional treatment group(even numbers), with 50 cases in each group.All patients received ultrasound-guided thoracoscopy to remove pleural effusion.The conventional treatment group received regular thoracic drainage catheters and the study group received pigtail catheters for drainage.Drainage was stopped after the patient was extubated.The levels of programmed cell death 1(PD-1), programmed death-ligand 1(PD-L1)and PD-1/PD-L1 in peripheral blood were detected and compared before and after catheterization; Values of clinical indicators including drainage volume, catheter retention time, length of hospitalization and complications during catheter drainage for the two groups were recorded.Results:After extubation, the levels of PD-1, PD-L1 and PD-1/PD-L1 in peripheral blood for both groups decreased, compared with pre-catheterization levels, with the study group having lower levels[(31.95±10.41)ng/L, (26.14±1.03)ng/L, (1.22±0.12)ng/L]than the conventional treatment group[(43.31±12.03)ng/L, (30.62±1.57)ng/L, (1.41±0.63)ng/L]( t=5.049, 16.871, 2.095, P<0.001, 0.039).The study group had a catheter retention time of(15.43±3.12)d and a length of hospitalization of(7.36±2.95)d, shorter than(11.32±3.87)d and(22.15±6.98)d in the conventional treatment group( t=5.754, 6.215, both P<0.001), but there was no statistical significant difference in drainage volume between the two groups( P>0.05).The incidence of drainage catheter falling out and catheter obstruction in the study group was lower than in the conventional treatment group[2.00%(1/50) vs.16.00%(8/50), 0.00%(0/50) vs.18.00%(9/50)]( χ2=4.396, 7.814, P=0.036, 0.005), and there was no statistical significant difference in the incidence of mediastinal shifting, subcutaneous emphysema, intrathoracic infections or hemothorax between the two groups(all P>0.05). Conclusions:Thoracoscopic ultrasound-guided pigtail catheter drainage for the treatment of elderly patients with tuberculous pleural effusion is clearly effective and may help to reduce the level of PD-1/PD-L1 in peripheral blood, shorten the time of catheter drainage and decrease some complications for patients.

Influence of the number of high-risk cytogenetic abnormalities on the clinical characteristics and prognosis in 360 patients with newly diagnosed multiple myeloma / 中华血液学杂志

Objective: To investigate the influence of the number of high-risk cytogenetic abnormalities (HRCA) on the clinical characteristics and prognosis of patients with newly diagnosed multiple myeloma (MM) . Methods: A total of 360 patients with newly diagnosed MM admitted to Jiangsu Province Hospital between November 2013 and September 2020 were included in this study. Cytoplasmic light chain immunofluorescence with fluorescence in situ hybridization (cIg-FISH) was used to detect HRCA. Cytogenetic abnormalities were combined with clinical characteristics and outcomes for further analysis. Results: Among the 360 patients, 120 patients (33.3%) presented with no HRCAs, and 175 (48.6%) , 61 (16.9%) , and four (1.1%) patients had one, two, and three HRCA (s) , respectively. Patients were divided into three groups, including the no-HRCA group, one-HRCA group, and ≥two-HRCA group, according to the number of HRCAs. There were significant differences in the R-ISS stage, hemoglobin level, albumin level, and the proportion of bone marrow plasma cells among the three groups (P<0.05) . The COX proportional-hazards model identified extramedullary disease (P=0.018) , HRCA ≥ 2 (P=0.001) , and absence of autologous hematopoietic stem cell transplantation (P<0.001) as independent risk factors for progression free survival (PFS) and identified lactate dehydrogenase (LDH) level ≥ 220 U/L (P<0.001) , HRCA ≥2 (P=0.001) , and absence of autologous hematopoietic stem cell transplantation (P=0.005) as independent risk factors for overall survival (OS) . The median PFS was 28 months, 22 months, and 14 months (P=0.005) for the three cohorts, and their OS was not reached,60 months, and 30 months (P=0.001) , respectively. Conclusions: HRCA ≥ 2 is an independent risk factor for decreased survival in patients with newly diagnosed MM. More HRCAs result in heavier tumor burden, as well as a higher risk of disease progression and death.

Exploration of four dimensional training path of adversity quotient for nursing undergraduates in medical colleges / 中华医学教育探索杂志

How to improve the ability of nursing undergraduates in medical colleges to face more difficulties and reverse the dilemma in response to public health emergencies is a topic that needs to be further studied. It is an effective way to improve the level of students' adversity quotient by cultivating students' adversity quotient. This paper expounds the four dimensions of the composition of the four-dimensional adversity quotient, analyzes the factors influencing the formation of the four-dimensional adversity quotient, and puts forward the exploration of the path and method to cultivate the four-dimensional adversity quotient of undergraduate nursing students. In the practical teaching of nursing specialty, the adversity quotient can be cultivated through listening and communication.

Research on the relationship between occupational cognition, coping style and transition shock of nursing interns / 中华医学教育探索杂志

Objective:To analyze the relationship between transition shock of nursing interns and occupational cognition and coping styles, and explore the factors affecting the status quo of nursing interns transformation, so as to provide guidance for the development of nursing students to respond to clinical transition shock.Methods:A cross-sectional survey of nursing students in Hunan Province was conducted among nursing interns from 18 colleges and universities in Hunan Province by using the General Demographic Data Questionnaire and the Transition Shock of Newly Graduated Nurses Scale, the Nurses' Professional Cognitive Survey Scale, and the Simplified Coping Style Questionnaire. The collected data were processed by SPSS 18.0 software, and multivariate linear regression analysis was performed to analyze the influencing factors of the shock.Results:The overall scores of nursing students transition shock were (64.49±16.33) points, while the transition shock was negatively correlated with occupational cognition ( r=-0.433, P<0.01) and positive response ( r=-0.263, P<0.01). And the occupational cognition, proposed academic qualification, reason for choosing nursing profession and positive response were the main factors affecting the transition shock of nursing students ( P=0.00, R2=0.21). Conclusion:The transition shock of nursing students is at the middle level, and has a strong negative correlation with the nursing students' occupational cognition and positive response. School teachers and clinical instructors should pay attention to the confusion, suspicion and unclear feelings and experiences caused by the transition shock of nursing students, helping nursing students to improve their occupational cognition, and learning how to positively respond to the transition shock problem, thereby improving the quality of clinical internship teaching and training of nursing students.

Application of the ligation clip-assisted modified " expansion and collapse method" in thoracoscopic resection of the external posterior basal segment in children with congenital pulmonary airway malformation / 中华实用儿科临床杂志

Objective:To explore the feasibility of using ligation clip-assisted modified " expansion and co-llapse method" to determine the intersegment plane in thoracoscopic resection of the external posterior basal segment (S 9+ 10) in children with congenital pulmonary airway malformation (CPAM). Methods:Retrospective study.The clinical data of 12 CPAM children who underwent thoracoscopic S 9+ 10 resection in Children′s Hospital Affiliated to Shandong University from July 2019 to May 2021 were collected and analyzed.There were 7 males and 5 females.The age at operation ranged from 3.50 to 11.50 months.The body weight of patients ranged from 6.5 to 11.5 kg.In all the patients, the ligation clip-assisted modified " expansion and collapse method" was applied during the operation to determine the intersegment plane (S 9+ 10 was in the expansion state and the remaining lungs were in the collapse state) before thoracoscopic S 9+ 10 resection.After S 9+ 10 resection, the surgical treatment and postoperative recovery were summarized and analyzed. Results:All the operations were completed under thoracoscopy, and there was no conversion to thoracotomy.In 11 patients, the intersegment plane was accurately determined by the ligation clips-assisted improved " expansion and collapse method" , and the S 9+ 10 segment was successfully resected.Of these 11 cases, 8 cases had right S 9+ 10 resection and 3 cases had left S 9+ 10 resection.In the process of using the ligation clip-assisted improved " expansion and collapse method" to determine the intersegment plane, the operator needed to clamp the ligation clip after the anesthesiologist expanded the lung completely.Because the operation was not stable enough, the ligation clip fell off and did not clamp S 9+ 10, resulting in operation failure.The operation was changed to the traditional " expansion and collapse method" . Besides, the basal segment bronchus was cut off by mistake, so the right basal segment had to be resected.The operation time ranged from 85 to 205 min, with a median of 149 min.Intraoperative bleeding ranged from 5 to 15 mL, with a median of 10 mL.The indwelling time of drainage tubes ranged from 2 to 4 days, with a median of 3 days.The postoperative hospital stay ranged from 5 to 8 days, with a median of 6 days.Postoperative pathological examination results suggested 4 cases of type 1, 6 cases of type 2 and 2 cases of type 3.There were 2 cases of simple subcutaneous emphysema without postoperative complications such as bleeding, bronchopleural fistula and atelectasis.The follow-up time ranged from 6 to 28 months, with a median of 15 months.The chest CT of all children showed no residual lesions and no residual cavities in the involved hemithorax. Conclusions:Ligation clip-assisted improved " expansion and collapse method" for determining the intersegment plane is suitable for CPAM children with a narrow thoracic space, and the operation is simple and effective.It is relatively safe and feasible to use this method in S 9+ 10 resection.

Fancd2os Reduces Testosterone Production by Inhibiting Steroidogenic Enzymes and Promoting Cellular Apoptosis in Murine Testicular Leydig Cells

Background@#It is well-established that serum testosterone in men decreases with age, yet the underlying mechanism of this change remains elusive. @*Methods@#The expression patterns of Fancd2 opposite-strand (Fancd2os) in BALB/c male mice and testicular tissue derived cell lines (GC-1, GC-2, TM3, and TM4) were assessed using real-time polymerase chain reaction (RT-PCR), Western blot and immunofluorescence. The Fancd2os-overexpressing or knockdown TM3 cells were constructed by infecting them with lentivirus particles and were used to evaluated the function of Fancd2os. The testosterone production was measured using enzyme linked immunosorbent assay (ELISA) and the steroidogenic enzymes such as steroidogenic acute regulatory protein (StAR), P450 cholesterol side-chain cleavage (P450scc), and 3β-hydroxysteroid dehydrogenase (3β-HSD) were analysed using RT-PCR. The apoptosis of TM3 cells induced by ultraviolet light or testicular tissues was detected using flow cytometry, Western blot or dUTP-biotin nick end labeling (TUNEL) assays. Pearson correlation analysis was used to assess the correlation between the Fancd2os expression and TUNEL-positive staining in mouse testicular Leydig cells. @*Results@#The Fancd2os protein was predominantly expressed in mouse testicular Leydig cells and its expression increased with age. Fancd2os overexpression inhibited testosterone levels in TM3 Leydig cells, whereas knockdown of Fancd2os elevated testosterone production. Fancd2os overexpression downregulated the levels of StAR, P450scc and 3β-HSD, while Fancd2os knockdown reversed this effect. Fancd2os overexpression promoted ultraviolet light-induced apoptosis of TM3 cells. In contrast, Fancd2os knockdown restrained apoptosis in TM3 cells. In vivo assays revealed that higher Fancd2os levels and mouse age were associated with increased apoptosis in Leydig cells and decreased serum testosterone levels. Pearson correlation analysis exhibited a strong positive correlation between the expression of Fancd2os and TUNEL-positive staining in mouse testicular Leydig cells. @*Conclusion@#Our findings suggest that Fancd2os regulates testosterone synthesis via both steroidogenic enzymes and the apoptotic pathway.

Diagnosis and differential diagnosis value of immune marker CCR7 in chronic lymphocytic leukemia and other chronic B-cell lymphoproliferative disorders / 中华检验医学杂志

Objective:To investigate the clinical significance of cc-chemokine receptor 7 (CCR7) as a potential diagnostic or differential marker for chronic lymphocytic leukemia (CLL).Methods:A total number of 643 patients with B-cell chronic lymphoproliferative diseases (B-CLPD) admitted to the First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2018 were enrolled. The patients included 327 cases of CLL, 58 cases of mantle cell lymphoma (MCL), 34 cases of follicular lymphoma (FL), 36 cases of marginal zone lymphoma (MZL), 10 cases of hair-cell leukemia or its variants (HCL/HCLV-v), 40 cases of Waldorf′s macroglobulinemia (WM), 48 cases of CD5 +B-cell chronic lymphoproliferative disease unclassified (B-CLPD-U) and 90 cases of CD5 -B-CLPD-U. At the same time, 20 samples from healthy people from the medical examination center of our hospital were used as normal controls. Flow cytometry was used to detect the immune-phenotype and CCR7 expression level in B-CLPD patients, and Fluorescence in situ hybridization (FISH) was used to analyze the genomic alterations: the ataxia telangiectasia mutant gene (ATM) deletion, the 13q14 deletion, the P53 deletion and trisomy 12. Sanger sequencing was used to analyze gene mutations of splicing factor 3B subunit 1 (SF3B1), NOTCH1, tumor protein 53 (TP53) and immunoglobulin heavy chain variable region (IGHV). Measurement data were compared by Mann-Whitney test, and the positive rates were compared by chi-square test. The diagnostic value and optimal positive cutoff value of CCR7 were calculated using receiver operating characteristic (ROC) curve. Results:The positive rates of CCR7 expression in typical CLL and atypical CLL were 90.8% (257/283) and 84.1% (37/44), respectively, and there was no significant difference of the positive rates (χ 2=1.228, P=0.268) between groups. The positive expression rates of CCR7 in CLL, MCL, CD5 +B-CLPD-U, CD5 -B-CLPD-U, FL, WM, HCL/HCL-v and MZL were 89.9% (294/327), 10.3% (6/58), 6.3% (3/48), 8.9% (8/90), 0, 0, 0 and 13.9% (5/36) respectively, and the median mean fluorescence intensity (MFI) was 278 (246, 307), 114 (106, 128), 112 (106, 117), 110 (104, 121), 108 (105, 119), 111 (105, 124), 112 (108, 115) and 109 (105, 120) respectively. Compared with CLL, the positive expression rates of CCR7 in other types of B-CLPDs were lower significantly (χ 2=181.3, 177.8, 232, 164.7, 180.8, 62.6, 129, P<0.01). In addition, the sensitivity, specificity and accuracy of CCR7 for distinguishing CLL from other types of B-CLPD were 89.9%, 93.0% and 92.3%, respectively. The positive expression rate of CD49d in CCR7 +CLL patients was 13.9%, which was significantly lower than that in CCR7 -CLL patients (42.1%) (χ 2=7.6, P=0.01). The coincidence rate of 13q14 deletion was 50.3% in CCR7 +CLL patients, which was significantly higher than that in CCR7 -CLL patients (20%) (χ 2=6.56, P=0.01). Conclusions:The CC-chemokine receptor 7 (CCR7) antigen is an effective marker for the diagnosis and identification of chronic lymphocytic leukemia (CLL). The expression level of CCR7 in clinical specimens can distinguish CLL from other pathological subtypes of B-CLPDs.

miRNA-34a reverses cisplatin resistance of osteosarcoma through targeted inhibition of high mobility group box 1 / 肿瘤研究与临床

Objective:To investigate the related mechanism of miRNA-34a (miR-34a) reverses cisplatin resistance of osteosarcoma through targeted inhibition of high mobility group box 1 (HMGB1).Methods:The MG-63 cisplatin-resistant cell line (MG-63/DDP) was constructed by using dose escalation and intermittent action, and then the successfully constructed MG-63/DDP cells were treated with different concentrations of cisplatin (0, 1, 5, 10, 20, 30 μg/ml), and CCK-8 method was used to detect cell survival rate. The MG-63/DDP cells were transfected respectively and then randomly divided into two groups: the miR-34a overexpression vector group and the miR-34a empty expression vector (miR-34a-NC) group. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of miR-34a. Transfected cells were treated with different concentrations of cisplatin (0, 1, 5, 10, 20, 30 μg/ml), and CCK-8 method was used to detect cell survival rate, flow cytometry was used to detect cell apoptosis. The dual luciferase reporter gene experiment was used to verify whether miR-34a regulated the expression of HMGB1, and Western blotting method was used to detect the HMGB1 protein expression level of the transfected cells. MG-63/DDP cells were transfected respectively and then randomly divided into two groups: HMGB1 gene silencing vector (si-HMGB1) group and its negative control vector (si-NC) group. Western blotting method was used to detect HMGB1 protein expression level and CCK-8 method was used to detect cell survival rate.Results:The MG-63/DDP cell line was successfully constructed. The survival rate of MG-63/DDP cells was higher than that of MG-63 cells when cells were treated with different concentrations of cisplatin (all P < 0.01), and half inhibitory concentration ( IC50) value of MG-63/DDP cells and MG-63 cells on cisplatin was 25.80 μg/ml and 0.47 μg/ml, respectively. qRT-PCR results showed that the relative expression level of miR-34a in MG-63/DDP cells was lower than that in MG-63 cells (0.46±0.04 vs. 1.02±0.05, t = 15.14, P < 0.01); compared with miR-34a-NC group, the relative expression of miR-34a in MG-63/DDP cells was increased in miR-34a overexpression vector group (1.67±0.09 vs. 1.00±0.02, t = -12.58, P < 0.01). Cell survival rate of miR-34a overexpression vector group and miR-34a-NC group was decreased with the increase in the concentration of cisplatin; cell survival rate of miR-34a overexpression vector group was lower than that of miR-34a-NC group when cells were treated with different concentrations of 5- 30 μg/ml cisplatin (all P < 0.01). The apoptotic rate of MG-63/DDP cells in miR-34a-NC group and miR-34a overexpression vector group was (25.1±1.7)% and (42.3±2.3)%, respectively when cells were treated with 20 μg/ml cisplatin; and in miR-34a overexpression vector group, MG-63/DDP cells had a higher rate of apoptosis ( P < 0.01). The dual luciferase reporter gene experiment results showed that compared with miR-34a-NC group, miR-34a overexpression vector group could inhibit the luciferase activity of PGL3- wild-type-HMGB1, and the difference was statistically significant ( t = 6.37, P < 0.01), while miR-34a overexpression vector group had no significant inhibitory effect on the luciferase activity of PGL3- mutant-HMGB1 ( t = 0.35, P = 0.74). The relative expression level of HMGB1 protein in miR-34a overexpression vector group was lower than that in miR-34a-NC group (0.43±0.02 vs. 1.00±0.14, t = 6.98, P < 0.01). Compared with si-NC group, the relative expression level and IC50 value of HMGB1 protein in si-HMGB1 group were reduced (all P < 0.05). Conclusion:Overexpression of miR-34a can enhance the chemosensitivity of osteosarcoma cells MG-63/DDP to cisplatin, and its mechanism may be related to the inhibition of HMGB1 expression.

Preliminary clinical observation on endoscopic longitudinal incision combined with bleomycin local injection for complex benign esophageal strictures / 中华消化内镜杂志

Data of 7 patients with complex benign esophageal strictures (CBESs) who underwent endoscopic longitudinal incision combined with local injection of bleomycin were retrospectively reviewed at Air Force Medical Center from August 2018 to November 2019.The length of preoperative esophageal stenosis was 4-14 cm and the minimum diameter of esophageal stenosis was 0.2-0.4 cm in 7 cases. The procedure was successful for all 7 patients.No adverse events occurred during 5-14 months of follow-up period.Restenosis and dysphagia occurred in 5 cases.The interval between the first endoscopic treatment and the recurrence of esophageal stenosis was 30-120 days.Among the 5 cases of recurrence, 4 cases remained unobstructed after 2 treatments and 1 case remained unobstructed after 4 treatments. The dysphagia scores of 7 patients were graded from 0 to 1 by the end of follow-up. Endoscopic longitudinal incision combined with bleomycin therapy in treatment of CBESs is safe and effective.

Correlation between serum high sensitivity C-reactive protein and carotid intima-media thickness / 中国综合临床

Objective:To investigate the correlation between serum high sensitivity C-reactive protein and carotid intima-media thickness.Methods:A total of 5 136 health examination subjects, aged ≥40 years old, who met the inclusion criteria and had complete data, were selected as the research objects.A unified questionnaire survey, blood biochemistry and carotid artery color doppler ultrasound examination were performed.According to the diagnostic criteria of hs-CRP published by American Heart Association (AHA), the subjects were divided into three groups: 0.05 mg/L<hs-CRP<1.00 mg/L, 1.00 mg/L≤ hs-CRP<3.00 mg/L, and hs-CRP≥ 3.00-10.00 mg/L.Stepwise linear regression, χ 2 trend test and stepwise regression analysis were used to explore the correlation between hs-CRP and CIMT. Results:(1)Five thousand one hundred and thirty-six patients were enrolled, including 3 048 males and 2 088 females, aged (54.87±11.65) years.(2) Taking CIMT as the dependent variable and hs-CRP (converted by Lg)as the independent variable, the stepwise linear regression analysis using the entry method showed that there was a linear correlation between hs-CRP and CIMT, and the standard regression coefficient was 1.107( P<0.01). (3) According to this study, CIMT was divided into two groups, CIMT≤ 0.84 mm group and CIMT>0.84 mm group.The results showed that when the concentration of hs-CRP was high, CIMT increased with the increase of hs-CRP( OR(95% CI) 1.24 (1.01~1.52), P<0.05). Conclusion:There was a positive correlation between hs-CRP and CIMT.Patients with higher levels of hs-CRP are more likely to develop CIMT thickening and increase the risk of arteriosclerotic disease.

Characteristics of implicit memory and related factors in patients with schizophrenia with negative and positive symptoms / 中华行为医学与脑科学杂志

Objective:To investigate the characteristics of implicit memory and its related factors in schizophrenic patients with negative and positive symptoms.Methods:Ninety-three schizophrenic patients (including 52 cases of negative symptoms group and 41 cases of positive symptoms group) and 30 normal controls (normal group) were tested with the method of Chinese character word completion method. The reaction time and correct rate were recorded and compared with analysis of variance and t-test, and the correlation with demographic factors was analyzed with Pearson correlation analysis. Results:Accuracy of implicit memory test in schizophrenia group: there were statistically significant differences in the accuracy of patients with different course of disease (≤5 years: (38±5)%, 5-15 years: (34±8)%, ≥15 years: (34±7)%, P<0.05).The differences were statistically significant in the accuracy of patients with different education levels(primary school: (35±6)%, junior and senior high school: (34±7)%, secondary college education and above: (39±5)%, P<0.05).The accuracy of patients with education years < 10 years ((34±7)%) was significantly lower than those with education years ≥10 years ((37±6)%, P<0.05).Reaction time results of implicit memory test in schizophrenia group : there were significant differences in reaction time of patients with different disease course(≤5 years: (3 248±971)ms, 5-15 years: (3 515±672)ms, ≥15 years: (3 925±842)ms, P<0.05).The differences were statistically significant in the reaction time of workers ((3 495±712)ms), farmers ((3 870±878)ms) and soldiers ((3 024±924)ms, P<0.05).The reaction time of patients with clozapine ((3 869±871)ms) was significantly higher than that of patients with olanzapine ((3 393±626)ms, P<0.05).Intergroup results of accuracy: the accuracy of normal control group ((40±5)%) was significantly higher than that of negative symptom group ((33±7)%, P<0.01).The accuracy of negative symptom group was significantly lower than that of positive symptom group ((37±6)%, P<0.01).Intergroup reaction time results: the reaction time of normal control group ((2 660±667)ms) was significantly lower than that of negative symptom group ((3 678±951)ms, P<0.01) and positive symptom group ((3 072±865)ms, P<0.05).The reaction time of negative symptom group ((3 678±951)ms)was significantly higher than that of positive symptom group( P<0.01).There was significant negative correlation between the accuracy of implicit memory and the course of disease in schizophrenia ( r=-0.22, P<0.05). Conclusion:The implicit memory of schizophrenic patients is related to the course of disease, taking drugs, and occupation.There may have differences in implicit memory between patients with negative and positive symptoms of schizophrenia.

Rhodiola crenulata extract decreases fatty acid oxidation and autophagy to ameliorate pulmonary arterial hypertension by targeting inhibiton of acylcarnitine in rats / 中国天然药物

Pulmonary arterial hypertension (PAH) is a devastating pulmonary circulation disease lacking high-efficiency therapeutics. The present study aims to decipher the therapeutic mechanism of Rhodiola crenulata, a well-known traditional chinese medicine with cardiopulmonary protection capacity, on PAH by exploiting functional lipidomics. The rat model with PAH was successfully established for first, following Rhodiola crenulata water extract (RCE) treatment, then analysis of chemical constituents of RCE was performed, additional morphologic, hemodynamic, echocardiographic measurements were examined, further targeted lipidomics assay was performed to identify differential lipidomes, at last accordingly mechanism assay was done by combining qRT-PCR, Western blot and ELISA. Differential lipidomes were identified and characterized to differentiate the rats with PAH from healthy controls, mostly assigned to acylcarnitines, phosphatidylcholines, sphingomyelin associated with the PAH development. Excitingly, RCE administration reversed high level of decadienyl-L-carnitine by the modulation of metabolic enzyme CPT1A in mRNA and protein level in serum and lung in the rats with PAH. Furthermore, RCE was observed to reduce autophagy, confirmed by significantly inhibited PPARγ, LC3B, ATG7 and upregulated p62, and inactivated LKB1-AMPK signal pathway. Notably, we accurately identified the constituents in RCE, and delineated the therapeutic mechansim that RCE ameliorated PAH through inhibition of fatty acid oxidation and autophagy. Altogether, RCE might be a potential therapeutic medicine with multi-targets characteristics to prevent the progression of PAH. This novel findings pave a critical foundation for the use of RCE in the treatment of PAH.

Morphological and Molecular Phylogenetic Data of the Chinese Medicinal Fungus Cordyceps liangshanensis Reveal Its New Systematic Position in the Family Ophiocordycipitaceae

A cordycipitoid fungus infecting Hepialidae sp. in Nepal was supposed to be identical to Cordyceps liangshanensis, originally described from southwestern China, and thus, transferred to the genus Metacordyceps or Papiliomyces in previous studies. However, our multi-gene (nrSSU-nrLSU-tef-1α-rpb1-rpb2) phylogenetic and morphological studies based on the type specimen and additional collections of C. liangshanensis revealed that the fungus belongs to the genus Ophiocordyceps (Ophiocordycipitaceae). Therefore, a new combination O. liangshanensis was made, and a detailed description of this species was provided.