Impact of hormone receptor status and tumor subtypes of breast cancer in young BRCA carriers.

BACKGROUND: Hormone receptor expression is a known positive prognostic and predictive factor in breast cancer; however, limited evidence exists on its prognostic impact on prognosis of young patients harboring a pathogenic variant (PV) in the BRCA1 and/or BRCA2 genes. PATIENTS AND METHODS: This international, multicenter, retrospective cohort study included young patients (aged ≤40 years) diagnosed with invasive breast cancer and harboring germline PVs in BRCA genes. We investigated the impact of hormone receptor status on clinical behavior and outcomes of breast cancer. Outcomes of interest [disease-free survival (DFS), breast cancer-specific survival (BCSS), and overall survival (OS)] were first investigated according to hormone receptor expression (positive versus negative), and then according to breast cancer subtype [luminal A-like versus luminal B-like versus triple-negative versus human epidermal growth factor receptor 2 (HER2)-positive breast cancer]. RESULTS: From 78 centers worldwide, 4709 BRCA carriers were included, of whom 2143 (45.5%) had hormone receptor-positive and 2566 (54.5%) hormone receptor-negative breast cancer. Median follow-up was 7.9 years. The rate of distant recurrences was higher in patients with hormone receptor-positive disease (13.1% versus 9.6%, P < 0.001), while the rate of second primary breast cancer was lower (9.1% versus 14.7%, P < 0.001) compared to patients with hormone receptor-negative disease. The 8-year DFS was 65.8% and 63.4% in patients with hormone receptor-positive and negative disease, respectively. The hazard ratio of hormone receptor-positive versus negative disease changed over time for DFS, BCSS, and OS (P < 0.05 for interaction of hormone receptor status and survival time). Patients with luminal A-like breast cancer had the worst long-term prognosis in terms of DFS compared to all the other subgroups (8-year DFS: 60.8% in luminal A-like versus 63.5% in triple-negative versus 65.5% in HER2-positive and 69.7% in luminal B-like subtype). CONCLUSIONS: In young BRCA carriers, differences in recurrence pattern and second primary breast cancer among hormone receptor-positive versus negative disease warrant consideration in counseling patients on treatment, follow-up, and risk-reducing surgery.

[Clinical observation on the treatment of ossification of the posterior longitudinal ligament of the cervical spine using 3D printed self-stable zero-profile artificial vertebral body].

Objective: To observe the clinical efficacy of a 3D printed self-stable zero-profile artificial vertebral body for anterior cervical corpectomy decompression and fusion in the treatment of ossification of the posterior longitudinal ligament (OPLL) of the cervical spine. Methods: In this prospective randomized controlled trial, patients diagnosed with OPLL in Luohe Central Hospital from January to July 2022 were divided into a zero-profile group (3D printed self-stable zero-profile artificial vertebral body was used for internal fixation and fusion after anterior cervical subtotal decompression,) and titanium-mesh group (titanium-mesh and titanium plate were used for internal fixation and fusion after anterior cervical subtotal decompression) according to envelope random method. Operation time, intraoperative blood loss, Japanese Orthopaedic Association (JOA) score and improvement rate, incidence of postoperative prosthesis subsidence, and bone graft fusion were recorded and compared between the two groups. Results: Finally, 21 patients in the zero notch group and 20 patients in the titanium mesh group were included in the study and were followed-up. In the zero-profile group, there were 16 males and 5 females, aged (48.0±12.7) years. In the titanium-mesh group, there were 14 males and 6 females, aged (49.8±10.2) years. All the 41 patients successfully completed the operation. In the zero-profile group, the surgical time was (50.04±8.45) minutes, the blood loss was (95.38±26.07) ml and the hospitalization cost was (42.32±6.12) thousand yuan. In the titanium-mesh group, the surgical time was (59.20±11.95) minutes, the blood loss was (93.10±27.86) ml and the hospitalization cost was (42.10±6.71) thousand yuan. The surgical time in the zero-profile group was shorter than that in the titanium-mesh group (P=0.007), and there was no statistically significant difference in blood loss and hospitalization costs between the two groups (both P>0.05). The 41 patients were followed-up for (14.29±1.45) months. Four cases (20.0%) in the titanium mesh group experienced swallowing difficulties and 0 cases in the zero incision group, the difference between the two groups was statistically significant (P=0.048). No intraoperative hematoma, spinal cord nerve recompression, airway crisis, incision infection complications, and no steel plate or screw breakage or displacement occurred after surgery. At the last follow-up, all cases had bone fusion. At the follow-up of 12 months after surgery, the JOA score of the zero incision group increased from preoperative (10.33±1.71) points to (15.47±0.81) points, with an improvement rate of 76.1%±15.7%; the JOA score of the titanium mesh group increased from (10.30±1.75) points to (15.30±0.92) points, with an improvement rate of 73.2%±16.7%; there was no statistically significant difference in improvement rate between the two groups (P=0.580). At the follow-up of 12 months after surgery, 1 case (4.8%) in the zero incision group and 8 cases (40.0%) in the titanium mesh group experienced implant sinking, and the difference between the two groups was statistically significant (P=0.009). Conclusion: Compared with titanium-mesh, 3D printed self-stable zero-profile artificial vertebral body for the treatment of OPLL of the cervical spine can achieve good surgical efficacy, shorter surgical time, lower incidence of postoperative chronic swallowing discomfort, and can provide a better bone material bonding interface and be less prone to prosthesis settlement.

[To determine the value of iMAX, a new electrodiagnostic method, and its application in the evaluation of peripheral motor nerve excitability].

Objective: To test the new method of iMAX (the minimum stimulus current that elicits the maximum compound muscle action potential amplitude) electrodiagnosis, verify the feasibility of this method in evaluating the excitability of peripheral motor axons, and preliminarily explore the clinical application value. Methods: This study was a cross-sectional study. A total of 50 healthy subjects were recruited from the outpatient department of Peking University Third Hospital from June 2022 to March 2023, including 25 males and 25 females, aged 25-68 (48±8) years. Eleven patients with Charcot-Marie-Pain-1A (CMT1A), 7 males and 4 females, aged 19-55 (41±13) years and 21 patients with diabetic peripheral neuropathy (DPN), 10 males and 11 females, aged 28-79 (53±16) years were enrolled in this study. iMAX of bilateral median nerves, ulnar nerves and peroneal nerves were detected in all patients. Repeatable motor responses with minimum motor threshold and amplitude of at least 0.1 mV and the minimum stimulus current intensity, at which the maximum compound muscle action potential amplitude is elicited, were measured respectively [1 mA increment is called (iUP) and, 0.1 mA adjustment is called (iMAX)].Comparison of the parameters: the parameters of threshold, iUP and iMAX were compared among different age groups, genders and sides, body mass index(BMI) values and detection time , as well as between CMT1A patients, DPN patients and healthy subjects. Results: In healthy subjects, the threshold, iUP value and iMAX value were (1.8±0.7) mA, (4.4±1.2) mA, and (4.2±1.3) mA respectively; ulnar nerve (3.1±1.6) mA, (6.8±3.2) mA, (6.4±3.2) mA; peroneal nerve (3.7±2.0) mA, (7.8±2.8) mA, (7.4±2.9) mA. There were statistically significant differences in threshold, iUP value and iMAX value among different age groups (all P<0.001).With the increase of age, there was a trend of increasing threshold, iUP, and iMAX values in different nerves, and the differences are statistically significant (all P<0.001). There were no significant differences in gender, side and detection time threshold, iUP value and iMAX value (all P>0.05). The parameters of healthy subjects with high BMI value were higher than those of healthy subjects with low BMI value(all P<0.05). Compared with the healthy subjects, the parameters of 11 CMT1A patients were significantly increased (all P<0.05), and the parameters of 21 DPN patients were slightly increased (P<0.05). Conclusion: The new iMAX method reflects the excitability of motor axons and early axonal dysfunction, which is an important supplement to the traditional nerve conduction, and can be used to monitor motor axon excitability disorders.

[Analysis of the status of excess heart age and its risk factors among residents aged 35 to 64 years in China].

Objective: To analyze the status of excess heart age and its risk factors among Chinese residents aged 35 to 64 years. Methods: The study subjects were Chinese residents aged 35 to 64 years who completed the heart age assessment by WeChat official account "Heart Strengthening Action" through the internet from January 2018 to April 2021. Information such as age, gender, body mass index (BMI), blood pressure, total cholesterol (TC), smoking history, and diabetes history was collected. The heart age and excess heart age were calculated according to the characteristics of individual cardiovascular risk factors and the heart aging was defined as excess heart age≥5 years and 10 years respectively. The heart age and standardization rate were calculated respectively based on the population standardization of the 7th census in 2021.CA trend test was used to analyze the changing trend of excess heart age rate and population attributable risk (PAR) was used to calculate the contribution of risk factors. Results: The mean age of 429 047 subjects was (49.25±8.66) years. The male accounted for 51.17% (219 558/429 047) and the excess heart age was 7.00 (0.00, 11.00) years. The excess heart age rate defined by excess heart age≥5 years and ≥10 years was 57.02% (the standardized rate was 56.83%) and 38.02% (the standardized rate was 37.88%) respectively. With the increase of the age and number of risk factors, the excess heart age rate of the two definitions showed an upward trend according to the result of the trend test analysis (P<0.001). The top two risk factors of the PAR for excess heart age were overweight or obese and smoking. Among them, the male was smoking and overweight or obese, while the female was overweight or obese and having hypercholesterolemia. Conclusion: The excess heart age rate is high in Chinese residents aged 35 to 64 years and the contribution of overweight or obese, smoking and having hypercholesterolemia ranks high.

[Advances in clinical application of electrical impedance tomography to evaluate pulmonary perfusion in critically ill patients].

Lung perfusion monitoring can provide accurate information on changes in pulmonary blood flow in critically ill patients, and thus help guide clinical diagnosis and treatment. However, due to inconveniences such as patient transport, conventional imaging techniques are unable to meet the demand for real-time monitoring of lung perfusion, more convenient and reliable real-time functional imaging techniques should be developed to optimise cardiopulmonary management in critically ill patients. Electrical impedance tomography (EIT) is a non-invasive, radiation-free, bedside functional imaging technique that can be used to assess lung perfusion in patients with acute respiratory distress syndrome, pulmonary embolisms and other conditions, thereby assisting with the diagnosis of disease, the adjustment of treatment protocols, and the assessment of treatment outcomes. In this review, we focused on advances in EIT for lung perfusion monitoring in critically ill patients.

[Testosterone levels in patients with varicocele and azoospermia].

OBJECTIVE: Androgen deficiency is common in aging males and may have unfavourable health consequences. Large-scale studies suggested low testosterone level might increse mortality and morbidity in ageing males. However, young men with low testosterone level might be neglected. Recent studies reported young men with infertility may have reduced testosterone level. To investigate the incidence of androgen deficiency in males with infertility and possible factors affecting the low testosterone level. METHODS: Between January 2011 and December 2012, 407 men with infertility caused by varicocele (VC), obstructive azoospermia (OA) and nonobstructive azoospermia (NOA) in our center were included. The number of men in each group of OA, NOA and VC was 141, 97 and 169, respectively. All the eligible patients underwent a serum testosterone assessment by a single morning blood draw (between 8:00 to noon) to test for concentration of the total testosterone. All serum samples were determined by radioimmunoassay in our andrology laboratory. Androgen deficiency was defined as having a total testosterone level less than 300 ng/dL. RESULTS: The mean age was (30.4±5.8) years. The mean testosterone level was (4.18±1.64) ng/dL (range 0.30 to 11.32 ng/dL). The overall incidence of androgen deficiency was 26.5% (108/407). The incidences of androgen deficiency in NOA, OA and VC groups were 40.2% (39/97), 19.1% (27/141) and 24.9% (42/169), respectively, which were significantly higher in the NOA than in the VC and OA groups (P < 0.001). The incidences had no difference between the VC and OA groups (P=0.229). Univariate analysis revealed the cause of infertility, FSH and the mean testis volume as possible affecting factors for androgen deficiency. However, on multivariate analysis the only cause of infertility was an independent predictor. The incidence of androgen deficiency was the highest in the NOA group [OR 0.492 (95% confidence interval 0.288-0.840)]. CONCLUSION: NOA and varicocele might be risk factors of androgen deficiency. Young men with NOA may have a higher possibility of low testosterone level. Testosterone level should be followed up after NOA and varicocele treatment. Androgen deficiency should be assessed in males with infertility in clinical practice.

[Clinicopathological festures and outcome of combined hepatocellular-cholangiocarcinoma].

Objective: To investigate the clinicopathological features and pathological types and outcome of combined hepatocellular-cholangiocarcinoma (cHCC-CC). Methods: 30 cases of cHCC-CC were collected from Jan 2007 to Jun 2021 at General Hospital of Southern Theatre Command, People's Liberation Army of China, and analyzed the clinicopathological features, immunohistochemistry and outcome. The histological morphology was classified according to the Goodman standard and the fifth edition of World Health Organization (WHO) classification of digestive system tumors. Results: According to the Goodman classification, 9 cases belonged to type Ⅰ (i.e., collision tumor), with a coincidental occurrence of hepatocellular carcinoma and cholangiocarcinoma in the same specimen, and 21 cases were Type Ⅱ or "transitional tumors", in which there were areas of intermediate differentiation and an identifiable transition between hepatocellular carcinoma and cholangiocarcinoma. According to the WHO classification of digestive system tumors (5th Edition), 25 cases were classified in cHCC-CC, and 5 cases were cHCC-CC-Intermediate cell carcinoma. There were 28 males and 2 females, with an average age of 50.4 (31-72) years old. 21 cases accompanied liver cirrhosis, with liver flukes in 1 case and HBsAg positive in 23 cases. Immunohistochemical staining showed nestin was positive in 9 cases, 66.7% died (6/9) and 33.3% (3/9) survived only 6 months. The 1-year recurrence rate was 3.9% for liver resection and 50% for orthotopic liver transplantation, and liver resection has longer median survival time than liver transplantation after recurrence. Conclusions: cHCC-CC is a rare type of primary liver malignant tumor. Preoperative diagnosis is difficult. The definite diagnosis depends on histopathological morphology and immunohistochemical markers. Nestin may be as a prognosis factor, and surgical treatment is preferably liver resection.

B Cell-Intrinsic Expression of Interferon Regulatory Factor 1 Supports Chronic Murine Gammaherpesvirus 68 Infection.

Gammaherpesviruses are ubiquitous pathogens that are associated with cancers, including B cell lymphomas. These viruses are unique in that they infect naive B cells and subsequently drive a robust polyclonal germinal center response in order to amplify the latent reservoir and to establish lifelong infection in memory B cells. The gammaherpesvirus-driven germinal center response in combination with robust infection of germinal center B cells is thought to precipitate lymphomagenesis. Importantly, host and viral factors that selectively affect the gammaherpesvirus-driven germinal center response remain poorly understood. Global deficiency of antiviral tumor-suppressive interferon regulatory factor 1 (IRF-1) selectively promotes the murine gammaherpesvirus 68 (MHV68)-driven germinal center response and expansion of the viral latent reservoir. To determine the extent to which antiviral effects of IRF-1 are B cell intrinsic, we generated mice with conditional IRF-1 deficiency. Surprisingly, B cell-specific IRF-1 deficiency attenuated the establishment of chronic infection and the germinal center response, indicating that MHV68 may, in a B cell-intrinsic manner, usurp IRF-1 to promote the germinal center response and expansion of the latent reservoir. Further, we found that B cell-specific IRF-1 deficiency led to reduced levels of active tyrosine phosphatase SHP1, which plays a B cell-intrinsic proviral function during MHV68 infection. Finally, results of this study indicate that the antiviral functions of IRF-1 unveiled in MHV68-infected mice with global IRF-1 deficiency are mediated via IRF-1 expression by non-B cell populations.IMPORTANCE Gammaherpesviruses establish lifelong infection in over 95% of all adults and are associated with B cell lymphomas. The virus's manipulation of the germinal center response and B cell differentiation to establish lifelong infection is thought to also precipitate malignant transformation, through a mechanism that remains poorly understood. The host transcription factor IRF-1, a well-established tumor suppressor, selectively attenuates MHV68-driven germinal center response, a phenotype that we originally hypothesized to occur in a B cell-intrinsic manner. In contrast, in testing, B cell-intrinsic IRF-1 expression promoted the MHV68-driven germinal center response and the establishment of chronic infection. Our report highlights the underappreciated multifaceted role of IRF-1 in MHV68 infection and pathogenesis.

[Identification of serum peptide biomarker for ovarian cancer diagnosis by Clin-TOF-II-MS combined with magnetic beads technology].

Objective: To explore the serum cyclic polypeptide biomarkers for ovarian cancer diagnosis. Methods: A total of 54 patients with epithelial ovarian cancer confirmed by pathology in Cancer Hospital, Chinese Academy of Medical Sciences from March 2018 to September 2018 were selected as the study subjects, and 40 healthy women with normal examination results in the cancer screening center were selected as the control. All of the samples were randomly divided into training set and validation set at the ratio of 1∶1 with a random number. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) combined with magnetic bead technology was used for detecting peptide profiling in serum samples to screen significantly differently expressed peptides between ovarian cancer group and control group of the training set (score>5). Receiver operating characteristic (ROC) curve analysis was used to screen differential peptide peaks with area under curve (AUC) ≥0.8, sensitivity and specificity>90% in the training set and validation set. Liquid chromatography-mass spectrometry (LC-MS/MS) was further used to determine the composition of differentially expressed peptides. Results: By comparing the peptide profiles of the two groups, 102 differential peptide peaks were initially detected in the mass-to-charge ratio range of 1 000 to 10 000. ROC curve analysis showed that there were 42 differential peptide peaks with AUC ≥0.8 in both training set and validation set, 19 of which were highly expressed in ovarian cancer group, and 23 were lowly expressed. There were 15 different peptide peaks in highly expressed ovarian cancer group with sensitivity and specificity over 90%. The mass-to-charge ratios were 7 744.27, 5 913.41, 5 329.87, 4 634.21, 4 202.02, 3 879.26, 3 273.35, 3 253.79, 3 234.34, 2 950.33, 2 664.51, 2 018.38, 1 893.37, 1 498.69 and 1 287.55. There were 15 different peptide peaks in lowly expressed ovarian cancer group with sensitivity and specificity over 90%, the mass-to-charge ratios were 9 288.46, 7 759.77, 5 925.24, 4 652.77, 4 210.42, 3 887.02, 3 279.90, 3 240.82, 2 962.15, 2 932.70, 2 022.42, 1 897.16, 1 501.69, 1 337.38 and 1 290.13. No protein composition was identified in 15 different peptide peaks in lowly expressed ovarian cancer group. The two protein compositions identified in 15 different peptide peaks in highly expressed ovarian cancer group were recombinant serglycin (SRGN) and fibinogen alpha chain (FGA), the mass-to-charge ratios of which were 1 498.696 and 5 913.417, respectively. The sensitivity and specificity of the two proteins for ovarian cancer diagnosis were 100%, 100% and 90.9%, 100%, respectively. Conclusion: SRGN and FGA are highly expressed in the serum of ovarian cancer patients, which may be potential diagnostic markers for ovarian cancer.

[Comparison of acquired immure deficiency syndrome-related knowledge and behavior between persons with and without human immunodeficiency virus- infection among young male students who have sex with men].

OBJECTIVE: To understand the differences between young male students who have sex with men (MSM) with and without human immunodeficiency virus (HIV)-infection in acquired immure deficiency syndrome (AIDS)-related knowledge and behavior, and to provide a scientific reference to make targeted and effective measures in AIDS prevention. METHODS: Using snow balling sampling combined with participants' referral, we conducted a questionnaire survey among 548 young MSM students (in whom there were both HIV-positive and HIV-negative) in Harbin, Tianjin, Xi'an, and Chongqing cities from April 2017 to March 2018. The chi-square test and binary Logistic regression were used to compare the differences in AIDS-related knowledge and behavior between males with and without HIV-infection. RESULTS: A total of 583 questionnaires were obtained, of which 548 were valid, with an effective rate of 94.0%. Having a junior college education or below (P=0.002), a monthly consumption level of less than 2 000 RMB (P=0.021), and living off campus (P=0.004) were associated with being tested positive for HIV. In any period of schooling, receiving AIDS prevention education was a protective factor for HIV infection [Primary school OR=0.203 (0.073－0.561), junior high school OR=0.287 (0.142－0.581), senior high school OR=0.271 (0.142－0.518), and university OR=0.322 (0.168－0.616)]. There was no statistical difference between HIV positive and negative young MSM students in the cognition of "AIDS-related Knowledge for Public"(P=0.907) and "AIDS-related Knowledge for Youth"(P=0.782), with the awareness rate all about 90%. There was a statistical difference in the need for some specific knowledge (For "AIDS prevention and treatment policy", P=0.012, for "Ways to identify and prevent high-risk sexual behavior", P < 0.001). HIV-positive males had a younger age of first sexual activity (P=0.006), had more sexual partners in the early (P < 0.001) and had lower frequency of condom use (P < 0.001). However, there was no statistical difference in the later number of sexual partners (P=0.247) and the frequency of condom use (For regular sex partners, P=0.735, and for casual sex partners, P=0.765), which might be related to the change of sexual behavior characteristics caused by HIV infection (For regular sex partners, P < 0.001, and for casual sex partners, P=0.006). CONCLUSION: There was no statistical difference between HIV positive and negative young MSM students in the cognition of "AIDS-related Knowledge for Public" and "AIDS-related Knowledge for Youth", which were both lower than 95% required by the state. However, the specificity in the knowledge needs was certainly shown. There was no significant difference in the recent sexual behavior between the two groups, but HIV positive students were more likely to have high-risk sexual behaviors in the early stage, so we should strengthen and move forward the sex education and AIDS prevention education with adjusted contents, and prevent high-risk sexual behaviors within young MSM students in the early stage.

[Association studies between postoperative hypoeosinophilia and hospital stay in patients undergoing hip fracture surgery under different anesthesia].

Objective: To investigate early postoperative changes in eosinophils (EOS) and the relationship of postoperative hospital stay and hypoeosinophilia in patients undergoing hip fracture surgery under general or spinal anesthesia. Methods: The clinical data of patients who underwent hip fracture surgery at Beijing Tiantan Hospital between April 2014 and November 2017 were retrospectively analyzed. Patients were classified according to whether they received general anesthesia or spinal anesthesia. Univariate regression analysis was used to examine a number of covariates potentially contributed to postoperative hospital stay. Multivariate linear regression was used to analyze the relationship between postoperative day 1 EOS counts and postoperative hospital stay. Interaction and stratified analyses were conducted according to anesthesia methods. Results: A total of 149 patients were included in this study. Thirty-four of them underwent general anesthesia and 115 of them underwent spinal anesthesia. The postoperative day 1 EOS of the general anesthesia group was 0 (0,1.8)×107/L, which was lower than that of the spinal anesthesia group 1.0(0,6.0)×107/L (Z=3.095, P<0.01). After adjusting the confounders of age, gender, American Society of Anesthesiologists (ASA) grade, intraoperative blood loss, intraoperative red blood cell (RBC) transfusion, postoperative day 1 hematocrit (HCT) and white blood cell (WBC), postoperative complications, and the interaction terms for ASA, intraoperative blood loss, intraoperative RBC transfusion and postoperative complications, the negative correlation between postoperative hospital stay and postoperative day 1 EOS level was significant (ß=-0.39, 95%CI:-0.74--0.05, P<0.05) in the general anesthesia group. Conclusion: EOS were significantly decreased after surgery in general anesthesia group, and the postoperative hospital stay was negatively correlated with postoperative day 1 hypoeosinophilia.

[Treatment of lumbar degenerative diseases with recapping laminoplasty and nerve root canal's decompression preserving the continuity of supraspinous ligament].

Objective: To explore the clinical effect of lumbar discectomy and nerve root canal's enlargement preserving the continuity of supraspinous ligament in the treatment of lumbar degenerative disease. Methods: The data of patients with lumbar degenerative disease who underwent operation from 2016 to 2018 were analyzed retrospectively, and the patients were divided into two groups according to the different operation. The treatment group (17 cases) was treated with recapping laminoplasty, lumbar discectomy and nerve root canal's enlargement, and the control group (28 cases) was treated with total laminectomy, nerve root canal's enlargement, lumbar discectomy, interbody fusion and internal fixation (PLIF). All patients were followed up for 12 to 27 months (mean 17.8 months). Japanese Orthopaedic Association Scores(JOA) and visual analogue scale(VAS) of pain were used to evaluate the clinical effect before and after the operation, lumbar dynamical X-ray and Cobb angle were collecting for imaging evaluation, and the adjacent segment degeneration at the last follow-up was recorded. Results: There was no significant difference in preoperative JOA score, VAS score and Lumbar Cobb angle between the two groups (all P>0.05). The operation time in the treatment group was shorter than that in the control group, and the blood loss during operation in the treatment group was lower than that in the control group, the bed rest time of the treatment group after operation was shorter than that in the control group ((79±14) vs (118±17) min, (151±38) vs (324±70) ml and (3.4±0.7) vs (4.3±1.0) d,respectively; t=-8.508, -10.724, -3.244, all P<0.01). In addition, compared with the control group, the volume of postoperative drainage in the treatment group also decreased significantly (t=-5.637, P<0.01). There was no significant difference in JOA score between the two groups 1 year after the operation (P>0.05), but there was significant difference in VAS score between the two groups, the treatment group was better than the control group (P<0.05). Compared with the control group, the lumbar Cobb angle in the treatment group increased significantly one year after the operation (55.3°±3.2° vs 38.4°±6.2°, t=10.391, P<0.05). During the follow-up, no loosening or fracture of the implants was found in all patients. Conclusion: Treatment of lumbar degenerative diseases with recapping laminoplasty and nerve root canal's decompression preserving the continuity of supraspinous ligament by ultrasound osteotome has the same clinical effect as PLIF. It has the advantages of shortening operation time, less bleeding, better maintenance of lumbar lordosis after operation and reduction of adjacent segment degeneration.

[Association of GMF-ß expression with Ki-67 and its significance in the prognostic evaluation of astrocytoma].

Objective: To clarify the correlation of the expression of glia maturation factor-ß (GMF-ß) with Ki-67 in astrocytoma, and to investigate the prognostic implications of combined detection of GMF-ß and Ki-67. Methods: One hundred and forty human astrocytoma samples (WHO Ⅱ-Ⅳ grade) were collected at Southwest Hospital, Army Medical University (the Third Military Medical University), China from 2006 to 2009. Clinicopathological information and 3-year follow-up data were collected. Expression of GMF-ß and Ki-67 was detected by single and double immunohistochemical staining, then the association of GMF-ß expression with Ki-67 and its significance in prognostic evaluation of astrocytoma were statistically analyzed. Results: GMF-ß expression in astrocytoma cells was correlated to both tumor grade and Ki-67 (both P<0.05); Kaplan-Meier survival analysis showed that GMF-ß and Ki-67 expression were negatively correlated to the 3 year-survival rates, respectively (both P<0.01). Further analysis demonstrated that the two factors were co-influenced on survival, showing a trend of "GMF-ßlow Ki-67low>GMF-ßhigh Ki-67low>GMF-ßlow Ki-67high>GMF-ßhigh Ki-67high" in 3-year survival rate with significant intergroup differences (P<0.05, P<0.01). Conclusions: GMF-ß expression is positively associated with Ki-67 in astrocytoma. Combined detection of GMF-ß and Ki-67 can predict prognosis of patients with glioma.

B Cell-Intrinsic SHP1 Expression Promotes the Gammaherpesvirus-Driven Germinal Center Response and the Establishment of Chronic Infection.

Gammaherpesviruses are ubiquitous pathogens that establish lifelong infections in the majority of adults worldwide. Chronic gammaherpesvirus infection has been implicated in both lymphomagenesis and, somewhat controversially, autoimmune disease development. Pathogenesis is largely associated with the unique ability of gammaherpesviruses to usurp B cell differentiation, specifically, the germinal center response, to establish long-term latency in memory B cells. The host tyrosine phosphatase SHP1 is known as a brake on immune cell activation and is downregulated in several gammaherpesvirus-driven malignancies. However, here we demonstrate that B cell- but not T cell-intrinsic SHP1 expression supports the gammaherpesvirus-driven germinal center response and the establishment of viral latency. Furthermore, B cell-intrinsic SHP1 deficiency cooperated with gammaherpesvirus infection to increase the levels of double-stranded DNA-reactive antibodies at the peak of viral latency. Thus, in spite of decreased SHP1 levels in gammaherpesvirus-driven B cell lymphomas, B cell-intrinsic SHP1 expression plays a proviral role during the establishment of chronic infection, suggesting that the gammaherpesvirus-SHP1 interaction is more nuanced and is modified by the stage of infection and pathogenesis.IMPORTANCE Gammaherpesviruses establish lifelong infection in a majority of adults worldwide and are associated with a number of malignancies, including B cell lymphomas. These viruses infect naive B cells and manipulate B cell differentiation to achieve a lifelong infection of memory B cells. The germinal center stage of B cell differentiation is important as both an amplifier of the viral latent reservoir and the target of malignant transformation. In this study, we demonstrate that expression of tyrosine phosphatase SHP1, a negative regulator that normally limits the activation and proliferation of hematopoietic cells, enhances the gammaherpesvirus-driven germinal center response and the establishment of chronic infection. The results of this study uncover an intriguing beneficial interaction between gammaherpesviruses that are presumed to profit from B cell activation and a cellular phosphatase that is traditionally perceived to be a negative regulator of the same processes.

Direct investigation of current transport in cells by conductive atomic force microscopy.

Currents play critical roles in neurons. Direct observation of current flows in cells at nanometre dimensions and picoampere current resolution is still a daunting task. In this study, we investigated the current flows in hippocampal neurons, PC12 cells and astrocytes in response to voltages applied to the cell membranes using conductive atomic force microscopy (CAFM). The spines in the hippocampal neurons play crucial roles in nerve signal transfer. When the applied voltage was greater than 7.2 V, PC12 cells even show metallic nanowire-like characteristics. Both the cell body and glial filaments of astrocytes yielded CAFM test results that reflect different electrical conductance. To our best knowledge, the electrical characteristics and current transport through components of cells (especially neurons) in response to an applied external voltage have been revealed for the first time at nanometre dimensions and picoampere current levels. We believe that such studies will pave new ways to study and model the electrical characteristics and physiological behaviours in cells and other biological samples.

Oral vaccination with the porcine circovirus type 2 (PCV-2) capsid protein expressed by Lactococcus lactis induces a specific immune response against PCV-2 in mice.

AIMS: Porcine circovirus type 2 (PCV2) can cause postweaning, multisystemic wasting syndrome in pigs, which leads to enormous losses in the swine industry worldwide. Here, a genetically engineered Lactococcus strain expressing the main protective antigen of PCV2, the Cap protein, was developed to act against PCV2 infection as an oral vaccine. METHODS AND RESULTS: Expression of the Cap protein was confirmed via western blot, ELISA and fluorescence microscopy. Over 90% of the recombinant pAMJ399-Cap/MG1363 survived a simulated gastrointestinal transit. It also survived the murine intestinal tract for at least 11 days. Then, the safety and immunogenicity of pAMJ399-Cap/MG1363 in orally immunized mice was evaluated. The levels of the sIgA, IgG and cytokines (IL-4 and IFN-γ) obtained from the mice immunized with pAMJ399-Cap/MG1363 were significantly higher than those in the control groups. CONCLUSIONS: pAMJ399-Cap/MG1363 can survive in the gastrointestinal transit and effectively induce mucosal, cellular and humoral immune response against PCV2 infection via oral administration. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the potential of the genetically engineered Lactococcus lactis as a candidate for an oral vaccine against PCV2.

[A novel trans-muscular approach for microsurgical bilateral decompression via unilateral approach for degenerative lumbar stenosis: a series study].

Objective: To compare clinical outcomes of microsurgical bilateral decompression via unilateral approach (MBDU) between trans longissimus and iliocostalis approach, a novel lateral paraspinal approach(LPA), and classic Wiltseapproach for single level degenerative lumbar stenosis. Methods: LPA approach was researched by imaging measurement and cadaver anatomy. Retrospective analysis of a total 124 cases(male/female 75/49, aged(55±14) years) received single-level MBDU due to degenerative lumbar stenosis from 2016 to 2018 in Beijing Tiantan Hospital. Patients were categorized into two groups according to spinal canal morphology. Group A: trefoil shape (n=48), received MBDU via LPA; Group B: round & oval shape (n=76), received MBDU via Wiltse approach. Clinical parameters, including visual analogue scale (VAS) of pain, OswestryDability Index (ODI), satisfaction rate were assessed before the operation and at the latest follow-up. The internal fixation and intervertebral fusion were evaluated by X-ray and CT.The data were compared with single sample t test between the two groups. Results: The feasibility of trans longissimus and iliocostalis approach was proved by imaging measurement and anatomic study. The tube's maximum tilt angle in LPA group was 65°, and it was 40°in Wiltse group. There was no significantly difference in the operating time, bleeding and drainage volume between the two groups((120±27) min vs (115±32) min, (104±31) ml vs (110±41) ml, (50±15) ml vs (47±18) ml, respectively, t=1.246,-1.917,1.730,all P>0.05). In both groups, VAS and ODI scores significantly improved at the latest follow-up when compared with those before the operation (all P<0.05), but those were all comparable between the two groups(all P>0.05). The excellent rate of subjective satisfaction for the clinical efficacy in group A and B was 89.6%(43/48) and 86.8%(66/76) respectively (χ(2)=0.208,P>0.05). Conclusions: LPA approachfor microsurgical bilateral decompression via unilateral approach (MBDU) in degenerative lumbar stenosis is safe and effective, it has advantages in direct vison of contralateral nerve root canal decompression for trefoil shaped spinal canal.

[Clinical safety and efficiency of ultrasonic bone curette used in anterior cervical discectomy and fusion surgery].

Objective: To investigate the safety and efficiency of ultrasonic bone curette used in anterior cervical discectomy and fusion surgery. Methods: As a retrospective study, we collected and analyzed the clinical data of 47 patients receiving anterior cervical discectomy and fusion surgery in Luohe Central Hospital from January 2014 to January 2017, there were 26 males and 21 females with a mean age of (52±9) years. According to the different surgical tools used in the process of decompression by resecting osteophytes or bone like an inverted Chinese character "" located at the posterior margin of the vertebral body, the patients were divided into two groups: ultrasonic bone curette group (group A) and traditional tools group (group B). The operating time, volume of intraoperative blood losing, complications, Japanese Orthopedic Association (JOA) score before and after the operation and improvement rate were recorded in the two groups. The t test was used to compare the data between the two groups. Results: In group A, the operating time was (47±7) min, blood loss was (49±4) ml, 1 case experienced urinary tract infection and there was no cerebrospinal fluid leakage or spinal cord injury. In group B, the operating time was (54±12) min and the blood loss was (117±16) ml, cerebrospinal fluid leakage occurred in 2 patients and the incision healed one-stage by local compression, hoarseness happened in 1 case and it disappeared after 2 weeks, 2 patients had swallowing discomfort and recovered in one month, no spinal cord injury occurred in this group. The operating time and blood loss in group A were lower than those in group B (t=2.691, 20.704, both P<0.05). And the incidence of complications in group A were lower than that in group B (χ(2)=4.157, P=0.041). The JOA score of group A at 3 days after surgery was improved for 39.0% when compared with that before the surgery, and it was improved for 71.6% at one year after the surgery. The JOA score in group B at 3 days after surgery was elevated for 38.7% from that before the surgery, and it increased for 69.4% at one year after the surgery. There was no significant different in JOA score before the surgery, 3 days and one year after the surgery between the two groups (t=0.611, 1.076, 0.061, all P>0.05). Conclusion: In the process of decompression by resecting osteophytes or bone located at the posterior margin of the vertebral body in the anterior cervical discectomy and fusion surgery, ultrasonic bone curette is safe and effective, and it can effectively shorten the operating time, decrease the blood loss and cut down the incidence of complications.

Construction of a doxycycline inducible lentivirus that expresses stem cell-specific miR-302 cluster.

INTRODUCTION: The polycistronic miR-302 cluster encodes five miRNA genes that have an important role in the regulation of embryonic stem cell function. Studies showed that the miR-302 cluster can reprogram both mouse and human fibroblasts to induced pluripotent stem cells (iPSCs) with high efficiency. The aim of this study was to generate an inducible lentivirus that expresses miR-302 cluster in order to further investigate somatic cell reprogramming by these miRNAs. MATERIALS AND METHODS: The miR-302 cluster was amplified by polymerase chain reaction technique from human genomic DNA and was ligated into pTRIPz, an inducible lentiviral vector. RESULTS: MRC5 fibroblasts and HEK293 (human embryonic kidney) cells were infected with pTRIPz-302 cluster lentivirus and the family of 302 miRNAs were strongly expressed in HEK293 cells but lowly expressed in MRC5 fibroblasts. When cultured in hESC conditions, MRC5 cells expressed only low levels of DNMT3B, Nanog, Oct4 and Lin28 and failed to show stem cell induction. The red fluorescent expression seen in the majority of MRC5 cells, indicated that the rate of infection by lentivirus was efficient. DISCUSSION: The efficiency of reprogramming may be improved perhaps by either using a different cell type or a high expression vector with a different type of promoter.

Examination of Alzheimer's disease by a combination of electrostatic force and mechanical measurement.

Alzheimer's disease (AD) is the most common dementia and neurodegenerative disorder disease in central nervous system, and threatens the people's health seriously. But till now there are no effective ways to diagnose and treat AD. Herein, the differences between charge propagation, surface morphology, and Young's modulus of the hippocampus, in healthy and AD mice have been investigated using advanced modes of Atomic Force Microscopy (AFM). The ability of charge transfer in AD samples has decreased compared with healthy samples. There are compact layered structures in hippocampus from healthy sample, whereas there are no obvious layered structures in hippocampus from AD sample. The Young's modulus of hippocampus from AD sample is about half of that from healthy sample. The results demonstrate that AFM will be helpful in the further study of the functions of hippocampus and pathogeny of AD. LAY DESCRIPTION: Alzheimer's disease is the most common cause of dementia, a neurodegenerative disease which especially affects elderly people, threatens people's health seriously. The pathological features of AD are senile plaques, neurofibrillary tangles, and cerebral atrophy. The main clinical symptoms of AD are progressive memory impairment, cognitive dysfunction, and loss of language skills, as there are disorder in limbic and cortices. The morbidity rate increases with age and is currently at 5% and 20% among people older than 65 and 85, respectively. Thus AD has a heavy socioeconomic burden, especially in an aging society. So far, there are no effective therapeutic clinical treatments, and no effective means and methods to test and diagnose early stage AD. This means that further studies are essential to gain a deeper, fundamental understanding of the pathogenesis of AD. In this work, the surface morphology, electrical and mechanical properties of the hippocampi from healthy and AD mice have been investigated by Atomic Force Microscopy (AFM) at nanometre scale for the first time. The hippocampi from healthy mice are composed of compact layered structures, whereas there are no obvious characteristic features in the hippocampi from AD mice. The electrical properties of hippocampus slices were studied by the Electrostatic Force Microscopy (EFM) mode of AFM. The charge propagation ability of axons from AD mice has decreased dramatically compared to that of healthy mice. After charge injection (injection voltage was +10 V) the charge density was 2.3 × 105 C µm-2 in healthy samples, which was larger than that from the AD samples (1.0 × 105 C µm-2 ). The Young's modulus of hippocampi from healthy and AD mice were 104.0 and 40.1 kPa, respectively, as revealed by the quantitative nanomechanical mapping (QNM) mode of AFM. It indicates that when AD occurs, the Young's modulus will be lower than the healthy sample. These observations and experimental approaches will be useful in future studies of the pathogeny of AD.