[Ferroptosis inducer IKE ameliorate pulmonary fibrosis in collagen-induced arthritis (CIA) mice via decreasing the expression of IL-6, CCL5 and CXCL9].

Objective To investigate the impact of imidazole ketone erastin (IKE), a ferroptosis inducer, on pulmonary fibrosis progression in mice with collagen-induced arthritis (CIA), and to understand its potential mechanism. Methods Chick type II collagen emulsified in complete Freund's adjuvant (CFA) was injected into DBA/1 mice, aged 8 to 10 weeks, to induce CIA. Fourteen days later, type II collagen emulsified in incomplete Freund's adjuvant (IFA) was administered to the mice. The mice were randomly divided into a control group, a CIA group and a CIA combined IKE group. The development of arthritis was monitored by evaluating the arthritis scores every two days until day 39 and then the mice were sacrificed for organ collection. The histopathological changes of joints were evaluated by HE staining, Safranin O-fast green staining and toluidine blue staining. The histopathological changes of organs including heart, liver, spleen, lung, and kidney were evaluated by HE staining, and Masson's trichrome staining was used to assess pulmonary fibrosis. The expression levels of smooth muscle actin α (α-SMA), fibroblast activating protein α (FAPα), transforming growth factor ß (TGF-ß), type I collagen (Col1), interleukin 1(IL-1), IL-6, IL-17 and tumor necrosis factor α (TNF-α) were detected by immunohistochemical staining. The expression levels of serum cytokines including IL-17α, IL-17F, TGF-ß1, ITG-ß6, TNF receptor superfamily menber 11B(TNFRSF11B), TNFRSF12A, IL-6, IL-1α, IL-1ß, IL-10, TNF-α, CCL5, CCL2, CXCL9, CXCL1, NADK, EPO, CSF2, TGF-α, CCL20 and CCL3 in serum were detected by Olink mouse exploratory panel. Results Histological staining in the CIA mice administered with IKE model demonstrated that IKE treatment reduced bone absorption and the degree of synovial inflammation when active inflammation was present. CIA mice administered with IKE showed lower expression levels of α-SMA, FAPα, TGF-ß, Col1, IL-1, IL-6, IL-17 and TNF-α, according to the immunohistochemical staining of the lung. In addition, the expression levels of CCL5, CXCL9 and IL-6 were also decreased in serum of CIA mice treated with IKE. Conclusion IKE not only ameliorates joint inflammation and bone damage, but also alleviates the inflammation and the progression of pulmonary fibrosis in CIA mice.

Co-immobilized multi-enzyme biocatalytic system on reversible and soluble carrier for saccharification of corn straw cellulose.

Herein, three enzymes (cellulase, ß-glucosidase, and pectinase) with synergistic effects were co-immobilized on the Eudragit L-100, and the recovery of co-immobilized enzymes from solid substrates were achieved through the reversible and soluble property of the carrier. The optimization of enzyme ratio overcomed the problem of inappropriate enzyme activity ratio caused by different immobilization efficiencies among enzymes during the preparation process of co-immobilized enzymes. The co-immobilized enzymes were utilized to catalytically hydrolyze cellulose from corn straw into glucose, achieving a cellulose conversion rate of 74.45% under conditions optimized for their enzymatic characteristics and hydrolytic reaction conditions. As a result of the reversibility and solubility of the carrier, the co-immobilized enzymes were recovered from the solid substrate after five cycles, retaining 54.67% of the enzyme activity. The aim of this study is to investigate the potential of co-immobilizing multiple enzymes onto the Eudragit L-100 carrier for the synergistic degradation of straw cellulose.

Enhanced recovery after surgery (ERAS) in sacral tumour surgery and comprehensive description of a multidisciplinary program: a prospective study in a specialized hospital in China.

PURPOSE: There were fewer data to guide the application of enhanced recovery after surgery (ERAS) theory into sacral tumour surgery. In the present study, we were aiming to describe a multidisciplinary program of ERAS and evaluate the availability in sacral tumour surgery. METHODS: This was a prospective study of patients with sacral tumour between March 2021 and September 2021 at a single centre. We proposed a multidisciplinary program of ERAS for pre-admission, preoperative, intraoperative, postoperative, and post-discharge clinical care which positively influenced outcomes of patients with sacral tumour. All patients were prospectively assigned into two groups, ERAS group in which patients received ERAS protocols (n = 63), No-ERAS group in which patients had conventional clinical pathways (n = 62). Patient data were collected which included demographics, preoperative preparation, detailed information of surgical procedure, 60-day reoperation rate, 60-day readmission, postoperative length of stay (PLOS), time to first ambulation and flatus after surgery, time to removal of last drainage tube, and visual analogue scale (VAS) score at first ambulation and discharge. Complications referred to ones that occurred within 60 days after surgery. The above parameters were compared between ERAS group and No-ERAS group. RESULTS: Time to first ambulation after surgery in ERAS group (mean 20.9 h) was significantly shorter than that in No-ERAS group (mean 104.3 ho). Meanwhile, time to first flatus after surgery in ERAS group (mean 26.7 h) was also significantly shorter than that in No-ERAS group (mean 37.3 h). Patients in ERAS group had statistically shorter PLOS (10.7 days) as compared to that in No-ERAS group (13.8 days). In ERAS group, 19 of 63 patients (30.2%) were discharged within seven days after surgery as compared to seven of 62 patients (11.3%) in No-ERAS group. VAS score at first ambulation in ERAS group was not obviously higher than that in No-ERAS group though the time of first ambulation in ERAS group was statistically earlier than one in No-ERAS group. Furthermore, VAS score at discharge in ERAS group was significantly lower than that in No-ERAS group. The rate of postoperative incision necrosis was 6.3% (4/63) in ERAS group and 8.1% (5/62) in No-ERAS group and all of these nine patients underwent reoperation before discharge. The difference was not statistically significant in the wound complication of incision necrosis and 60-day reoperation rate. Only one readmission occurred in No-ERAS group due to the surgical site infection and also there was no significant difference of 60-day readmission rate between these two groups. Furthermore, there was no statistical difference of complications of femoral artery thrombosis and rectal rupture between ERAS group and No-ERAS group. CONCLUSIONS: Our proposed ERAS pathway for sacral tumour surgery and early walking facilitate safe and prompt discharge. ERAS protocols of sacral tumour surgery could decrease PLOS without significantly increasing postoperative complications, 60-day readmission rate and 60-day reoperation rate. The application of ERAS pathway in the field of sacral tumour surgery should have personalized feature with regard to resection type.

An optimized "sTOP" strategy-based awake fiberoptic intubation for a patient with severe scoliosis after halo-pelvic traction.

Difficult Airway Society launched the new guideline for awake tracheal intubation (ATI) in adults with the goal of standardizing and promoting ATI techniques to protect the airway in 2020 (Anaesthesia, 2020;75:509). Specifically, the guideline highlighted that the key components of ATI are sedation, topicalization, oxygenation, and performance, coined "sTOP." To the best of our knowledge, anticipated difficult airway is the best indication for ATI. Patients with severe scoliosis undergoing halo-pelvic traction (HPT) are often with head and neck fixation, thereby contributing to the anticipated difficult airways. HPT was first used to fix unstable cervical vertebra segments in 1959, and gradually applied in the treatment of scoliosis (scoliosis or kyphosis Angle greater than 90 degrees is usually considered as severe scoliosis), with favorable efficacy and safety profile, and thus widely used in clinical practice (Clin Orthop Relat Res, 1973;93:179). To date, the improved HPT device usually consists of a head ring composed of 6 ~ 8 cranial nails, a pelvic ring composed of 6 ~ 8 iliac bone nails and 4 telescopic connecting rods, which can achieve all-day continuous traction. Usually, the average traction time was about 8 weeks (Chin Med J (Engt), 2012;125:1297). Our case described a planned awake fiberoptic intubation (AFOI) for a patient with severe scoliosis undergoing HPT via an optimized "sTOP" strategy.

CD147 contributes to SARS-CoV-2-induced pulmonary fibrosis.

COVID-19 patients can develop clinical and histopathological features associated with fibrosis, but the pathogenesis of fibrosis remains poorly understood. CD147 has been identified as a universal receptor for SARS-CoV-2 and its variants, which could initiate COVID-19-related cytokine storm. Here, we systemically analyzed lung pathogenesis in SARS-CoV-2- and its delta variant-infected humanized CD147 transgenic mice. Histopathology and Transmission Electron Microscopy revealed inflammation, fibroblast expansion and pronounced fibrotic remodeling in SARS-CoV-2-infected lungs. Consistently, RNA-sequencing identified a set of fibrosis signature genes. Furthermore, we identified CD147 as a crucial regulator for fibroblast activation induced by SARS-CoV-2. We found conditional knockout of CD147 in fibroblast suppressed activation of fibroblasts, decreasing susceptibility to bleomycin-induced pulmonary fibrosis. Meplazumab, a CD147 antibody, was able to inhibit the accumulation of activated fibroblasts and the production of ECM proteins, thus alleviating the progression of pulmonary fibrosis caused by SARS-CoV-2. In conclusion, we demonstrated that CD147 contributed to SARS-CoV-2-triggered progressive pulmonary fibrosis and identified CD147 as a potential therapeutic target for treating patients with post-COVID-19 pulmonary fibrosis.

Inhibition of ferroptosis and iron accumulation alleviates pulmonary fibrosis in a bleomycin model.

Idiopathic pulmonary fibrosis (IPF) is a chronic progressive disease characterized by excessive proliferation of fibroblasts and excessive accumulation of extracellular matrix (ECM). Ferroptosis is a novel form of cell death characterized by the lethal accumulation of iron and lipid peroxidation, which is associated with many diseases. Our study addressed the potential role played by ferroptosis and iron accumulation in the progression of pulmonary fibrosis. We found that the inducers of pulmonary fibrosis and injury, namely, bleomycin (BLM) and lipopolysaccharide (LPS), induced ferroptosis of lung epithelial cells. Both the ferroptosis inhibitor liproxstatin-1 (Lip-1) and the iron chelator deferoxamine (DFO) alleviated the symptoms of pulmonary fibrosis induced by bleomycin or LPS. TGF-ß stimulation upregulated the expression of transferrin receptor protein 1 (TFRC) in the human lung fibroblast cell line (MRC-5) and mouse primary lung fibroblasts, resulting in increased intracellular Fe2+, which promoted the transformation of fibroblasts into myofibroblasts. Mechanistically, TGF-ß enhanced the expression and nuclear localization of the transcriptional coactivator tafazzin (TAZ), which combined with the transcription factor TEA domain protein (TEAD)-4 to promote the transcription of TFRC. In addition, elevated Fe2+ failed to induce the ferroptosis of fibroblasts, which might be related to the regulation of iron export and lipid metabolism. Finally, we specifically knocked out TFRC expression in fibroblasts in mice, and compared with those in the control mice, the symptoms of pulmonary fibrosis were reduced in the knockout mice after bleomycin induction. Collectively, these findings suggest the therapeutic potential of ferroptosis inhibitors and iron chelators in treating pulmonary fibrosis.

Risk Factors for Generalization in Patients with Ocular Myasthenia Gravis: A Multicenter Retrospective Cohort Study.

INTRODUCTION: Many patients with ocular myasthenia gravis (OMG) progress to generalized disease within the first 2 years of the onset of ocular symptoms. Several retrospective studies have identified risk factors associated with generalization, however these studies included patients on immunosuppression therapy or those undergoing thymectomy, which may reduce the generalization risk. In this study we explored the risk factors for generalization in non-immunosuppressed and non-thymectomized patients with OMG. METHODS: Data from patients with OMG treated at seven tertiary hospitals in China were retrospectively reviewed. Clinical characteristics, including sex, age at onset, symptoms at onset, comorbid autoimmune diseases, neostigmine test response, repetitive nerve stimulation (RNS) findings, presence of serum anti-acetylcholine receptor antibody (AChR-Ab), and thymic status based on radiological and pathological studies, were collected. The main outcome measure was disease generalization. The follow-up period was defined as the date of ocular symptom onset to the date of confirmation of generalization or immunotherapy initiation, or last follow-up (defined as 60 months). The Cox proportional hazards model was used to assess the risk factors for generalization. RESULTS: Overall, 572 patients (269 women) were eligible for inclusion in the analysis, of whom 144 developed generalization. The mean (standard deviation) onset age was 45.5 (19.8) years, and the median (interquartile range) follow-up period was 14.5 (7.0-47.3) months. Multivariable Cox regression analysis demonstrated that both early-onset (adjusted hazard ratio [aHR] 5.34; 95% confidence interval [CI] 1.64-17.36; p = 0.005) and late-onset (aHR 7.18; 95% CI 2.22-23.27; p = 0.001) in adulthood, abnormal RNS findings (aHR 3.01; 95% CI 1.97-4.61; p < 0.001), seropositivity for AChR-Ab (aHR 2.58; 95% CI 1.26-5.26; p = 0.01), and thymoma (aHR 1.62; 95% CI 1.05-2.49; p = 0.03) were independently associated with increased risk of generalization. CONCLUSION: The risk of generalization increased significantly in patients with adult-onset OMG, abnormal RNS findings, seropositivity for AChR-Ab, and thymoma, suggesting that these risk factors may predict OMG generalization.

NMN recruits GSH to enhance GPX4-mediated ferroptosis defense in UV irradiation induced skin injury.

Oxidative stress and lipid peroxidation are major causes of skin injury induced by ultraviolet (UV) irradiation. Ferroptosis is a form of regulated necrosis driven by iron-dependent peroxidation of phospholipids and contributes to kinds of tissue injuries. However, it remains unclear whether the accumulation of lipid peroxides in UV irradiation-induced skin injury could lead to ferroptosis. We generated UV irradiation-induced skin injury mice model to examine the accumulation of the lipid peroxides and iron. Lipid peroxides 4-HNE, the oxidative enzyme COX2, the oxidative DNA damage biomarker 8-OHdG, and the iron level were increased in UV irradiation-induced skin. The accumulation of iron and lipid peroxidation was also observed in UVB-irradiated epidermal keratinocytes without actual ongoing ferroptotic cell death. Ferroptosis was triggered in UV-irradiated keratinocytes stimulated with ferric ammonium citrate (FAC) to mimic the iron overload. Although GPX4 protected UVB-injured keratinocytes against ferroptotic cell death resulted from dysregulation of iron metabolism and the subsequent increase of lipid ROS, keratinocytes enduring constant UVB treatment were markedly sensitized to ferroptosis. Nicotinamide mononucleotide (NMN) which is a direct and potent NAD+ precursor supplement, rescued the imbalanced NAD+/NADH ratio, recruited the production of GSH and promoted resistance to lipid peroxidation in a GPX4-dependent manner. Taken together, our data suggest that NMN recruits GSH to enhance GPX4-mediated ferroptosis defense in UV irradiation-induced skin injury and inhibits oxidative skin damage. NMN or ferroptosis inhibitor might become promising therapeutic approaches for treating oxidative stress-induced skin diseases or disorders.

CD147 antibody specifically and effectively inhibits infection and cytokine storm of SARS-CoV-2 and its variants delta, alpha, beta, and gamma.

SARS-CoV-2 mutations contribute to increased viral transmissibility and immune escape, compromising the effectiveness of existing vaccines and neutralizing antibodies. An in-depth investigation on COVID-19 pathogenesis is urgently needed to develop a strategy against SARS-CoV-2 variants. Here, we identified CD147 as a universal receptor for SARS-CoV-2 and its variants. Meanwhile, Meplazeumab, a humanized anti-CD147 antibody, could block cellular entry of SARS-CoV-2 and its variants-alpha, beta, gamma, and delta, with inhibition rates of 68.7, 75.7, 52.1, 52.1, and 62.3% at 60 µg/ml, respectively. Furthermore, humanized CD147 transgenic mice were susceptible to SARS-CoV-2 and its two variants, alpha and beta. When infected, these mice developed exudative alveolar pneumonia, featured by immune responses involving alveoli-infiltrated macrophages, neutrophils, and lymphocytes and activation of IL-17 signaling pathway. Mechanistically, we proposed that severe COVID-19-related cytokine storm is induced by a "spike protein-CD147-CyPA signaling axis": Infection of SARS-CoV-2 through CD147 initiated the JAK-STAT pathway, which further induced expression of cyclophilin A (CyPA); CyPA reciprocally bound to CD147 and triggered MAPK pathway. Consequently, the MAPK pathway regulated the expression of cytokines and chemokines, which promoted the development of cytokine storm. Importantly, Meplazumab could effectively inhibit viral entry and inflammation caused by SARS-CoV-2 and its variants. Therefore, our findings provided a new perspective for severe COVID-19-related pathogenesis. Furthermore, the validated universal receptor for SARS-CoV-2 and its variants can be targeted for COVID-19 treatment.

Safety and efficacy of meplazumab in healthy volunteers and COVID-19 patients: a randomized phase 1 and an exploratory phase 2 trial.

Recent evidence suggests that CD147 serves as a novel receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Blocking CD147 via anti-CD147 antibody could suppress the in vitro SARS-CoV-2 replication. Meplazumab is a humanized anti-CD147 IgG2 monoclonal antibody, which may effectively prevent SARS-CoV-2 infection in coronavirus disease 2019 (COVID-19) patients. Here, we conducted a randomized, double-blinded, placebo-controlled phase 1 trial to evaluate the safety, tolerability, and pharmacokinetics of meplazumab in healthy subjects, and an open-labeled, concurrent controlled add-on exploratory phase 2 study to determine the efficacy in COVID-19 patients. In phase 1 study, 59 subjects were enrolled and assigned to eight cohorts, and no serious treatment-emergent adverse event (TEAE) or TEAE grade ≥3 was observed. The serum and peripheral blood Cmax and area under the curve showed non-linear pharmacokinetic characteristics. No obvious relation between the incidence or titer of positive anti-drug antibody and dosage was observed in each cohort. The biodistribution study indicated that meplazumab reached lung tissue and maintained >14 days stable with the lung tissue/cardiac blood-pool ratio ranging from 0.41 to 0.32. In the exploratory phase 2 study, 17 COVID-19 patients were enrolled, and 11 hospitalized patients were involved as concurrent control. The meplazumab treatment significantly improved the discharged (P = 0.005) and case severity (P = 0.021), and reduced the time to virus negative (P = 0.045) in comparison to the control group. These results show a sound safety and tolerance of meplazumab in healthy volunteers and suggest that meplazumab could accelerate the recovery of patients from COVID-19 pneumonia with a favorable safety profile.

Prophylactic temporary abdominal aortic balloon occlusion for patients with pernicious placenta previa: a retrospective study.

BACKGROUND: Pernicious placenta previa (PPP) can increase the risk of perioperative complications. During caesarean section in patients with adherent placenta, intraoperative blood loss, hysterectomy rate and transfusion could be reduced by interventional methods. Our study aimed to investigate the influence of maternal hemodynamics control and neonatal outcomes of prophylactic temporary abdominal aortic balloon (PTAAB) occlusion for patients with pernicious placenta previa. METHODS: This was a retrospective study using data from the Peking University People's Hospital from January 2014 through January 2020. Clinical records of pregnant women undergoing cesarean section were collected. Patients were divided into two groups: treatment with PTAAB placement (group A) and no balloon placement (group B). Group A was further broken down into two groups: prophylactic placement (Group C) and balloon occlusion (group D). RESULTS: Clinical records of 33 cases from 5205 pregnant women underwent cesarean section were collected. The number of groups A, B, C, and D were 17, 16, 5 and 12.We found that a significant difference in the post-operative uterine artery embolism rates between group A and group B (0% vs.31.3%, p = 0.018). There was a significant difference in the Apgar scores at first minute between group A and group B (8.94 ± 1.43 vs 9.81 ± 0.75,p = 0.037),and the same significant difference between two groups in the pre-operative central placenta previa (29.4% vs. 0%,p = 0.044), complete placenta previa (58.8% vs 18.8%, p = 0.032),placenta implantation (76.5% vs 31.3%, p = 0.015). We could also observe the significant difference in the amount of blood cell (2.80 ± 2.68vs.10.66 ± 11.97, p = 0.038) and blood plasma transfusion (280.00 ± 268.32 vs. 1033.33 ± 1098.20, p = 0.044) between group C and group D. The significant differences in the preoperative vaginal bleeding conditions (0% vs 75%, p = 0.009), the intraoperative application rates of vasopressors (0% vs. 58.3%, p = 0.044) and the postoperative ICU (intensive care unit) admission rates (0% vs. 58.3%, p = 0.044) were also kept. CONCLUSIONS: PTAAB occlusion could be useful in reducing the rate of post-operative uterine artery embolism and the amount of transfusion, and be useful in coping with patients with preoperative vaginal bleeding conditions, so as to reduce the rate of intraoperative applications of vasopressors and the postoperative ICU (intensive care unit) admission. In PPP patients with placenta implantation, central placenta previa and complete placenta previa, we advocate the utilization of prophylactic temporary abdominal aortic balloon placement.

Effect of Self-Controlled Exercise on Antioxidant Activity of Red Blood Cells and Functional Recovery of Limbs in Patients with Breast Cancer after Rehabilitation.

BACKGROUND: We aimed to investigate the effect of self-controlled exercise on the antioxidant activity of red blood cells and the recovery of limb function in patients with breast cancer after rehabilitation. METHODS: Overall 130 breast cancer patients admitted to the First Affiliated Hospital of Zhengzhou University, China from Feb 2018 to Jan 2019 were divided into intervention group and control group. The control group received perioperative care and chemotherapy, the intervention group implemented a self-controlled exercise program. Indexes were compared between the two groups before intervention, 3 months and 6 months after intervention. RESULTS: The activity of erythrocyte superoxide dismutase (SOD) in the intervention group was significantly increased in the first 3 months (P=0.030), and decreased from 3rd to 6th month (P=0.033). The glutathione peroxidase (GSH-Px) activity in the intervention group increased through the whole intervention period. The plasma malondialdehyde (MDA) in the intervention group was significantly decreased (P=0.029, 0.012). After intervention for 3 months and 6 months, the 6MND distances in the intervention group were significantly longer (P=0.001, 0.045). The average exercise time in the intervention group were significantly increased (P=0.004, 0.000). CONCLUSION: Self-controlled exercise can effectively improve the antioxidant ability of red blood cells in patients with breast cancer, improve the mobility of shoulder joints of the affected side and increase their exercise capacity, with good sustainability. It has positive effect on postoperative rehabilitation, could be used in long-term regular clinical work.

Oncogenic Activation of YAP Signaling Sensitizes Ferroptosis of Hepatocellular Carcinoma via ALOXE3-Mediated Lipid Peroxidation Accumulation.

Ferroptosis, a form of programmed cell death process driven by iron-dependent lipid peroxidation, plays an important role in tumor suppression. Although previous study showed that intracellular Merlin-Hippo signaling suppresses ferroptosis of epithelial tumor cells through the inactivation of YAP signaling, it remains elusive if the proto-oncogenic transcriptional co-activator YAP could serve as a potential biomarker to predict cancer cell response to ferroptosis-inducing therapies. In this study, we show that both total YAP staining and nuclear YAP staining were more prevalent in HCC tissues than in nontumorous regions. Compared to low-density HCC cells, high-density cells showed decreased nuclear localization of YAP and conferred significant resistance to ferroptosis. Oncogenic activation of YAP signaling by overexpression of YAP(S127A) mutant sensitized ferroptosis of HCC cells cultured in confluent density or in the 3D tumor spheroid model. Furthermore, we validated the lipoxygenase ALOXE3 as a YAP-TEAD target gene that contributed to YAP-promoted ferroptosis. Overexpression of ALOXE3 effectively increased the vulnerability of HCC cells to ferroptotic cell death. In an orthotopic mouse model of HCC, genetic activation of YAP rendered HCC cells more susceptible to ferroptosis. Finally, an overall survival assay further revealed that both a high expression of YAP and a low expression of GPX4 were correlated with increased survival of HCC patients with sorafenib treatment, which had been proven to be an inducer for ferroptosis by inhibition of the xc-amino acid antiporter. Together, this study unveils the critical role of intracellular YAP signaling in dictating ferroptotic cell death; it also suggests that pathogenic alterations of YAP signaling can serve as biomarkers to predict cancer cell responsiveness to future ferroptosis-inducing therapies.

Research Advances in Chemokine-like Factor Super Family Member 8.

Chemokine-like factor super family member (CMTM) is a novel generic family firstly reported by Peking University Center for Human Disease Genomics. CMTM8 is one member of this family and has exhibited tumor-inhibiting activities. It can encode proteins approaching to the transmembrane 4 superfamily. CMTM8 is down-regulated in most carcinoma cell lines and tissues. Over-expression of CMTM8 may inhibit the proliferation,migration,and invasion of carcinoma cells. However,the exact mechanism of its anti-tumor activity remains unclear. CMTM8 may be involved in various signaling pathways governing the occurrence and development of tumors. CMTM8 may be a new target in the gene therapies for tumors,while further studies on CMTM8 and its anti-tumor mechanisms are warranted.

Cryopreservation of stallion spermatozoa using different cryoprotectants and combinations of cryoprotectants.

The present study investigates the effects of five cryoprotectants (CPAs) and cryoprotectant combinations on the post-thaw total motility, progressive motility, viability, mitochondrial membrane potential and acrosome integrity in stallion spermatozoa. In Experiment 1, the objective was to compare the impact of different concentrations (2.5%, 3.5% and 5%) of a single CPA, including glycerol (Gly), ethylene glycol (EG), dimethyl sulphoxide (DMSO), methyl formamide (MF), and dimethylformamide (DMF) for stallion spermatozoa cryopreservation. In Experiment 2, two or more CPAs were used to assess whether this improved post-thaw spermatozoa quality. Gly, MF and DMF, were used to prepare seven combinations of freezing extender with different mixtures of cryoprotectant, and the 3.5% Gly, MF and DMF were used as a control group. The results show that post-thaw total motility, progressive motility, viability, and mitochondrial membrane potential for all concentrations of EG and DMSO were less than the 3.5% and 5% Gly and MF and 2.5% and 3.5% DMF (P<0.05). Use of the 3.5% concentration resulted in the greater post-thaw total motility and progressive motility than the 2.5% and 5% concentrations for all CPAs. The results for the use of different combinations of cryoprotectant indicate there are differences in progressive motility and viability. The viability with the use of Gly(2/3)+MF(1/3) was 44.65% and was greater than the Gly(1/3)+MF(1/3)+DMF(1/3) (30.96%), MF(2/3)+DMF(1/3) (35.05%), Gly (32.21%) and MF(33.76%) (P<0.05). The progressive motility with the use of the MF(2/3)+Gly(1/3) combination was 36.0% and was greater than in the DMF (25.0%) and MF(2/3)+DMF(1/3) (22.7%) (P<0.05). These results suggest that using the appropriate cryoprotectant combination instead of a single cryoprotectant can improve horse spermatozoa cryopreservation.

[Expression of brainderived neurotrophic factor in urine of patients with benign prostatic hyperplasia complicated overactive bladder symptoms].

OBJECTIVE: To investigate the expression of urinary brainderived neurotrophic factor (BDNF) in benign prostatic hyperplasia patients with overactive bladder (OAB) symptoms and its correlation with the severity of OAB symptoms. METHODS: According to the inclusion and exclusion criteria, a total of 178 patients with benign prostatic hyperplasia who were to undergo transurethral resection of prostate (TURP) were enrolled in this study. All the patients had accepted basic preoperative evaluations, as well as an assessment of their International Prostate Symptom Score (IPSS) and Overactive Bladder Symptom Score (OABSS). The patients who had been scheduled for surgery had to take the urodynamic assessment. Urinary BDNF levels were measured by the enzyme-linked immunosorbent assay (ELISA) and the results were further normalized to the concentration of urinary creatinine (BDNF/Cr, mg/mol). RESULTS: The urinary BDNF/Cr levels of the patients with moderate and severe lower urinary tract symptoms were (1.189 ± 0.753) mg/mol and (1.817 ± 1.110) mg/mol (P < 0.001). The urinary BDNF/Cr levels of the patients with grades III-VI obstruction were (1.382 ± 0.945) mg/mol, (1.435 ± 0.938) mg/mol, (1.640 ± 1.104) mg/mol, and (1.653 ± 1.019) mg/mol, respectively (P > 0.05). There was no correlation between the urinary BDNF/Cr levels and the severity of obstruction (r = 0.103, P = 0.173). The urinary BDNF/Cr levels in the patients with and without OAB symptoms were (1.913 ± 0.843) mg/mol and (0.297 ± 0.183) mg/mol (P < 0.001). The urinary BDNF/Cr levels in the patients with mild, moderate and severe OAB symptoms were (1.501 ± 0.543) mg/mol, (1.806 ± 0.703) mg/mol and (2.560 ± 0.979) mg/mol, respectively (P < 0.05). There was a correlation between the urinary BDNF/Cr levels and the severity of OAB symptoms (r = 0.743, P < 0.001). The urinary BDNF/Cr levels in the patients with urodynamic detrusor overativity were significantly higher than those without detrusor overativity [(1.917 ± 0.866) mg/mol and (1.194 ± 1.013) mg/mol, P < 0.001]. CONCLUSION: There is no correlation between urinary BDNF and severity of obstruction in benign prostatic hyperplasia patients with moderate and severe lower urinary tract symptoms. The urinary BDNF levels in patients with OAB symptoms are elevated compared with patients without OAB symptoms, and are correlated with the severity of OAB symptoms.

Nanoparticle-to-vesicle and nanoparticle-to-toroid transitions of pH-sensitive ABC triblock copolymers by in-to-out switch.

An efficient way to achieve nanoparticle-to-vesicle transition of ABC triblock copolymers by in-to-out switch of the pH-sensitive core-forming C block is described.

[Study on cytogenetic changes with relation to FAB classification in 397 patients with acute leukemias].

The purpose of study was to investigate the cytogenetic abnormality of acute leukemias (AL), to analyze the relationship in the chromosomal abnormality and the AL FAB types, and to explore the impact of the chromosomal abnormalities on the prognostic factors of AL. The chromosome karyotypes of 397 patients with AL were analyzed by means of bone marrow short-term culture and G banding technique. The results showed that in 319 out of 397 patients, the chromosome karyotypes could be analyzed, and the chromosomal abnormality occurred in 175 patients (54.9%). In the patients with acute lymphocytic leukemia (ALL), acute myelogenous leukemia (AML) and acute mixed-lineage leukemia (AMLL), the chromosomal abnormality occurred respectively in 33 of 120 patients (27.5%), 129 of 252 patients (51.2%) and 13 of 25 patients (52.0%). Hyper-diploids, hypo-diploids and diploids occurred in 41 of 175 patients (23.4%), 22 of 175 patients (12.5%), and 112 of 175 patients (64.0%) respectively. In patients with AML the FAB type-associated chromosomal abnormality occurred in 69 of 129 patients (53.5%). It is concluded that chromosomal abnormalities exist in about 55% AL patients. Some special chromosomal abnormalities are cytogenetic characteristics of AL, and obviously correlated with AL FAB types, the combination of chromosomal detection with cytogenetics is useful for the diagnosis of AL, and the evaluation of therapeutic effects and prognosis.

Preparation and characterization of nanoaggregates self-assembled by lithocholic acid and N-trimethyl modified chitosan derivatives.

A novel kind of chitosan derivatives (CS-LA-TM) were synthesized by grafting hydrophobic molecules of lithocholic acid (LA) and quaternization. CS-LA-TM and micelle-like self-aggregates were characterized by FTIR, 1H NMR, fluorescence spectroscopy, dynamic light scattering, and transmission electron microscopy (TEM). The critical micelles concentration (CMC) ranging from 0.009 mg/mL to 0.030 mg/mL decreased with increasing of the degree of substitution (DS) of LA, pH of medium but with decreasing of the degree of quaternization (DQ) of amino groups. The TEM images demonstrated that spherical CS-LA-TM nanoaggregates with uniform size were formed by self-assembly. The sizes (100-200 nm) of CS-LAs-TMs nanoaggregates increased with increasing of DS, DQ, and can be easily controlled by pH of medium, which may confer the nanoaggregates potential as delivery systems for anticancer drugs, or DNA and siRNA.

[Sensitivity of interferon-gamma release assay T-SPOT. TB in diagnosing extrapulmonary tuberculosis].

OBJECTIVE: To evaluate the sensitivity of an interferon-gamma release assay T-SPOT. TB in the diagnosis of bacteriologically or histologically confirmed extrapulmonary tuberculosis. METHOD: Totally 31 patients with bacteriologically or histologically confirmed extrapulmonary tuberculosis in Peking Union Medical College Hospital received T-SPOT. TB assay to detect early secreting antigen target 6 or culture filtrate protein 10 peptides-specific T cells in the peripheral blood mononuclear cells (PBMCs). RESULTS: T-SPOT. TB assay showed positive results in 29 patients with extrapulmonary tuberculosis and the sensitivity was 93.5% (95% CI 84.8% - 100%). The median of spot forming cells (SFCs) in response to early secreting antigen target 6 peptides was 196/10(6) PBMCs (interquartile range, 72-532/10(6) PBMCs), the median of SFCs in response to culture filtrate protein 10 peptides was 276 SFCs/10(6) PBMCs (interquartile range, 72-568/10(6) PBMCs), and the median of the incorporate SFCs was 612/10(6) PBMCs (interquartile range, 192-1 152/10(6) PBMCs). CONCLUSION: T-SPOT. TB is highly sensitive in diagnosing extrapulmonary tuberculosis.