Effects of miR-143 on the migration and invasion of osteosarcoma cells by regulating MMP-13 expression / 中国骨伤

OBJECTIVE@#To explore the effect of miR-143 regulating matrix metalloproteinase(MMP)-13 expression on migration and invasion of osteosarcoma cells.@*METHODS@#The mouse osteosarcoma cell line 143B cells were cultured in 96-well plates, and blank group, negative group, positive group, and intervention group were set up. Then, the blank group did no treatment 50 μg miR-143 mimic was added to positive group, negative group added equal mimic NC (control sequence of miR-143 mimic), the intervention group was added 50 μg miR-143 mimic and 10 μg MMP-13 protein, all groups continued to culture for 3 to 6 hours, and finally the serum was aspirated to treat for half an hour. The protein expressions of miR-143 and MMP-13 in each group were measured by fluorescence quantitative PCR experiment and Western blot experiment, respectively, and the invasion and migration abilities of cells were measured by Transwell and scratch experiments.@*RESULTS@#The expression of MMP-13 protein in the positive group and the intervention group was significantly lower than that in the blank group, and the positive group was lower than the intervention group (P<0.05);The mean numbers of invasive cells in blank group, negative group, positive group and intervention group were (1 000.01±44.77), (959.25±46.32), (245.04±4.33), (634.06±33.78) cells/field, respectively;the scratch healing rate of the positive group and the intervention group was significantly lower than that of the blank group, and the positive group was lower than the intervention group (P<0.05).@*CONCLUSION@#MMP-13 is a target of miR-143, which can reduce the migration and invasion ability of osteosarcoma cells by inhibiting the expression of MMP-13.

Short-term clinical outcomes of adult cardiac surgery in patients with prior COVID-19 in a single center / 中国胸心血管外科临床杂志

@#Objective     To provide experience for clinical diagnosis and treatment through exploring the perioperative characteristics and short-term treatment outcomes of adult cardiac surgery in patients with prior coronavirus disease-2019 (COVID-19). Methods    A retrospective analysis was performed on patients undergoing coronary artery bypass grafting (CABG) or valve surgery in the Department of Cardiac Surgery of Beijing Anzhen Hospital from December 26, 2022 to December 31, 2022, and previously diagnosed with COVID-19 before surgery. Results    Finally 108 patients were collected, including 81 males and 27 females, with an average age of 60.73±8.66 years. Two (1.9%) patients received emergency surgery, and the others received elective surgery. The 86.1% of patients had been vaccinated, and the duration of COVID-19 was 5.0 (4.0, 7.0) days. The time from COVID-19 to operation was 15.0 (12.0, 17.8) days. Eighty-nine patients received CABG, of which off-pump CABG was dominant (92.1%). Nineteen patients received valve surgery. The rate of delayed extubation of ventilator was 17.6%. The ICU stay was 21.0 (17.3, 24.0) hours, and the postoperative hospital stay was 7.0 (6.0, 8.0) days. Three (2.8%) patients were treated with intra-aortic balloon  pump (IABP), one (0.9%) patient was treated with extracorporeal membrane oxygenation (ECMO), one (0.9%) patient was treated with continuous renal replacement therapy (CRRT) due to acute renal insufficiency, three (2.8%) patients were treated with temporary pacemaker, and one (0.9%) patient underwent rethoracotomy. In terms of postoperative complications, the incidence of cerebrovascular accident, acute renal insufficiency, gastrointestinal bleeding and septicemia was 0.9%, respectively, and the incidence of acute heart failure, lung infection, and liver insufficiency was 1.9%, respectively. All patients recovered and were discharged from hospital, and no in-hospital death occurred. Conclusion     The utilization rate of postoperative IABP, ECMO, CRRT, temporary pacemaker and the incidence of serious complications in patients with prior COVID-19 are not higher than those of normal patients, and the short-term treatment outcome is good.

Application of ventricular assist devices after heart failure: history and prospect / 中华胸心血管外科杂志

The treatment of heart failure is a common clinical problem faced by the medical community all over the world. It is also the only cardiovascular disease in the world that is on the rise. Although drug therapy for heart failure has made great progress in recent years, there are still many problems. Therefore, reversing ventricular remodeling, assisting or even replacing the function of the damaged heart and improving the prognosis and quality of life of patients with heart failure through surgical treatment will become the main battlefield for the future treatment of heart failure. As a transitional support therapy or permanent replacement therapy before heart transplantation, ventricular assist device can significantly improve the survival and quality of life of patients with heart failure, and is becoming an important treatment method for patients with end-stage heart failure. This paper aim to review the history and prospect of ventricular assist devices after heart failure.

Short-term and long-term outcomes of tricuspid valve replacement with mechanical and bioprosthetic valves / 中华胸心血管外科杂志

Objective:To investigate the difference between the short-term and long-term survival rates of patients undergoing tricuspid valve replacement with biological and mechanical valves.Methods:273 patients who received tricuspid valve replacement were selected from our Hospital from November 1993 to August 2018. The mean follow-up time was(8.2±5.6) years. The total follow-up rate was 95%. Kaplan-Meier method was used to make survival curves of the two groups and log rank test was used to compare the differences between the two groups. In addition, this study made the comparison of preoperative, intraoperative and postoperative information and long-term survival rate between these two groups.Results:There was no significant difference in demographic characteristics and baseline between mechanical valve group and biological valve group. 16 patients died in the mechanical valve group and 22 in the biological valve group. In the mechanical valve group, 14 cases died of postoperative low cardiac output syndrome and 2 cases died of gastrointestinal hemorrhage. 22 patients were died of low cardiac output syndrome. The auxiliary time in the mechanical valve group was longer than that in the biological valve group[(151.76±70.30)min vs.(131.62±60.25)min, P=0.013)]. There was no significant difference in long-term survival rate between the two groups in Kaplan- Meier survival curve( P=0.234). Conclusion:There is no difference in short-term and long-term survival rate between mechanical valve and biological valve in tricuspid valve replacement.

Alkali extraction of Ganoderma lucidum β-glucan and its anti-tumor immune regulation / 药学学报

In this study, an immunostimulating particulate β-glucan was isolated from a hot alkaline extract of the fruiting bodies of Ganoderma lucidum. The optimum conditions of 8 hours treatment time, 1∶20 solid - liquid ratio and 55 ℃ for the alkaline extract process were obtained after investigating by single-factor experiments and Box-Benhnken design in terms of the Ganoderma lucidum particulate β-glucan (GLG) increment, and these conditions resulted in a GLG yield of 8.57%. The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Jiangsu Provincial Academy of Chinese Medicine. The result showed that resident macrophages were effectively activated by GLG, such as with the up-regulation of co-stimulatory molecules, the secretion of cytokines and phagocytic uptake. GLG could also promote the proliferation of spleen lymphocytes in mice. In addition, IFN-γ production of spleen CD4+ T cells and cytotoxic T lymphocyte (CTL) responses were significantly enhanced on GLG orally treatment, which ultimately resulted in significantly decreased tumor burden. Taken together, these data suggest that GLG might act as an immune stimulator to exert antitumor effects.

Short-term and long-term outcomes of off-pump coronary artery bypass grafting in patients aged 80 years and over / 中华老年医学杂志

Objective:To investigate short-term and long-term outcomes of off-pump coronary artery bypass grafting(OPCABG)in patients aged 80 years and over.Methods:In this retrospective study, clinical data were collected, from June 2005 to July 2014, of 31 patients aged 80 and over years with left ventricular dysfunction who had received OPCABG in Beijing Anzhen Hospital and a follow-up of 3-12 years after discharge.Results:Ages of the patients ranged from 80 to 88 years with a mean age of(82.0±2.1)years.There were 25 males(80.6%). The mean left ventricular ejection fraction(LVEF)of patients was(42.0±6.5)% before operation, and 15 patients had LVEF less than 35%(48.4%). The average operation time was 4 hours.Three patients died in hospital(9.7%), of whom, one died from acute respiratory failure and the two others from low cardiac output syndrome.Perioperative complications included myocardial infarction(1 case, 3.2%), acute cerebral infarction(2 cases, 6.5%), tracheal intubation(3 cases, 9.7%), secondary thoracotomy and hemostasis(1 case, 3.2%), continuous renal replacement therapy(3 cases, 9.7%)and perioperative intra-aortic balloon pump(IABP)(8 cases, 25.8%). Twenty-eight patients were followed up for 3-12 years(median, 8.0 years), and no one had withdrawn.There were 10 all-cause deaths(35.7%), including 5 cardiogenic deaths(17.9%), 2 lung cancer deaths(7.1%), 2 deaths due to sepsis from systemic infection(7.1%), and 1 death from unknown causes.Besides, 2 patients had re-admission(7.1%), 2 patients suffered angina(7.1%)and 1 patient(3.6%)had recurrent myocardial infarction(3.6%). No patient received re-revascularization.Coronary computed tomography angiography(CTA)was performed on all surviving patients 1 year after surgery, and the patency rate of vascular bridges was 100%.Conclusions:Both short-term and long-term clinical outcomes of OPCABG are fair in patients aged over 80 years with left ventricular dysfunction, but the mortality and risk of complications are still high, and comprehensive preoperative evaluation is needed for these patients.

UHPLC-HRMS metabonomics to study effect of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats / 中国中药杂志

The project was launched to analyze the effects of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats by metabonomics. The preparation method of sulfur-fumigated Ophiopogonis Radix in laboratory was established. Then the blood samples of SD rats in blank group,Ophiopogonis Radix extract group and sulfur-fumigated Ophiopogonis Radix extract group were investigated by UHPLC-Q-Exactive. The differential metabolites were screened and identified by PCA(principal component analysis),OPLSDA(orthogonal partial least squares discriminant analysis) and variable importance projection(VIP),and the metabolic pathways were analyzed. Finally,a total of 15 potential biomarkers were identified. Compared with the samples of Ophiopogonis Radix extract group,sulfur-fumigated Ophiopogonis Radix mainly affected the biosynthesis and metabolism of amino acids in normal rats. Its mechanism may be related to the biosynthesis of phenylalanine,tyrosine,tryptophan and aminoacyl-tRNA as well as the metabolism of phenylalanine and tryptophan. Based on UHPLC-HRMS metabonomics,this paper discussed the effects of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats,which provided an idea for the metabolic study of other sulfur-fumigated traditional Chinese medicines.

Danshen: a phytochemical and pharmacological overview / 中国天然药物

Danshen, the dried root or rhizome of Salvia miltiorrhiza Bge., is a traditional and folk medicine in Asian countries, especially in China and Japan. In this review, we summarized the recent researches of Danshen in traditional uses and preparations, chemical constituents, pharmacological activities and side effects. A total of 201 compounds from Danshen have been reported, including lipophilic diterpenoids, water-soluble phenolic acids, and other constituents, which have showed various pharmacological activities, such as anti-inflammation, anti-oxidation, anti-tumor, anti-atherogenesis, and anti-diabetes. This article intends to provide novel insight information for further development of Danshen, which could be of great value to its improvement of utilization.

Inductive effect of solamargine on the apoptosis of human esophageal cancer KYSE150 cells and its action mechanism / 医学研究生学报

Objective Solamargine (SM), with its anti-inflammatory and anti-tumor effects, inhibits the proliferation and promotes the apoptosis of various tumor cells. This study was to investigate the effects of SM on the proliferation and apoptosis of human esophageal cancer KYSE150 cells and its action mechanism. Methods We treated KYSE150 cells with SM at the concentrations of 0 (the blank control group), 2, 4, 6 and 8 μmol/L for 24 hours. Then, we observed the morphological changes of the cells under the inverted microscope, detected their proliferation and apoptosis by MTT assay and flow cytometry respectively, and determined the expressions of the classical NF-κB signaling pathway-related proteins NF-κB, p-NF-κB, IKKα, IKKβ, IkBα and p-IkBα) and apoptosis-related proteins Bax, caspase-3, cleaved caspase-3 and Bcl-2 in different groups of the cells by Western blot. Results Compared with the blank control, the inhibition rate of the proliferation of the KYSE150 cells in the 2, 4, 6 and 8 μmol/L SM groups was increased significantly in a concentration-dependent manner (0 vs [15.03 ± 0.15]%, [47.94 ± 1.74]%, [68.72 ± 0.47]% and [77.51 ± 1.70]%, P<0.05), and so was the apoptosis rate ([8.17 ± 0.51]% vs [14.50 ± 0.73]%, [18.57 ± 2.08]%, [65.10 ± 10.88]% and [81.55 ± 5.48]%, P<0.05). The expression of the apoptosis-related protein Bax in the SM treated cells was up-regulated, those of Bcl-2, IKKα, IKKβ and p-IkBα down-regulated, and the activity of caspase-3 and cleaved caspase-3 promoted, all in a concentration-dependent manner, with statistically significant differences between the blank control and the 4, 6 and 8 μmol/L SM groups (P<0.05). Statistically significant differences were also found in the expressions of NF-κB, p-NF-κB and IkBα between the blank control and the 6 and 8 μmol/L SM groups (P<0.05). Conclusion Solamargine can significantly inhibit the proliferation and promote the apoptosis of KYSE150 cells, probably by suppressing the classical NF-κB signaling pathway.

The inhibition effects of allicin on TGF-β1 induced epithelial⁃mesenchymal transition of human cholangiocarcinoma cell and its mechanism / 医学研究生学报

Objective　The metastasis mechanism of cholangiocarcinoma is complex, which may be related to epithelial-mesenchymal transition(EMT). This study focused on investigating the inhibition effects of allicin on TGF-β1 induced epithelium mesenchymal transition of human cholangiocarcinoma cells and its related mechanism, and providing theoretical basis for the application of allicin in the treatment of cholangiocarcinoma.　Methods　MTT assay were used to detect the inhibition effects of different concentrations of allicin on the human cholangiocarcinoma RBE cell proliferation, and the drug concentration of allicin was determined by IC50 of 24 h. The RBE cells were cultured and divided into control group, allicin group(130.7μmol/L), TGF-β1 group(10ng/mL) and allicin+ TGF-β1 group(130.7μmol/L+10ng/mL). Wound scratch and transwell invasion assay were performed to detect the migration and invasion ability of RBE cells after 24 hours. Western blots were applied to detect expression of EMT-related proteins (E-Cadherin, N-Cadherin, Vimentin, Snail) and NF-κB signaling pathways.　Results　The migration rates in allicin group and allicin+ TGF-β1 group were both decreased compared with that in the control group ( 9.25% ± 0.36% vs 28.19 %±0.66%, P<0.05) and TGF⁃β1 group(13.91%±0.75% vs 49.22%±0.27%, P<0.05). The invasion rates in allicin group and allicin+ TGF-β1 group were also decreased compared with that in the control group (6.59%±0.06% vs 33.48%±0.04%, P<0.05) and TGF⁃β1 group(9.4%± 0.05% vs 40.21%±0.12%, P<0.05). Compared with the control group, E-Cadherin expression was significantly increased, and N-Cadherin, Vimentin, Snail, NF-κB and p-NF-κB expression were significantly decreased in the allicin group (P<0.05). Compared with TGF-β1 group, E-Cadherin expression was significantly up-regulated, and N-Cadherin, Vimentin, Snail, NF-κB and p-NF-κB expression were significantly down-regulated in the allicin+ TGF-β1 group (P<0.05)．　Conclusion　These results indicate that allicin can inhibit the EMT induced by TGF-β1 on the human cholangiocarcinoma cell by blocking NF-κB signaling pathway, which may have potential value to be the drug candidate for the treatment of human cholangiocarcinoma in future.

Intracranial hemorrhage from metastatic CNS lymphoma: A case report and literature review

@#Metastatic brain lymphomas, which belong to secondary central nervous system lymphomas, usually originate from primary tumors of the bone marrow, testis, or orbit. Gastrointestinal lymphomas commonly metastasize to the lung or heart. We report here a case of brain hemorrhage due to metastasis from primary gastrointestinal diffuse large B-cell lymphoma (DLBCL). A 30-year-old male presented with headache. He was diagnosed to have gastrointestinal lymphoma 6 months earlier, and treated with gastrointestinal surgery. Pathological diagnosis was DLBCL. A PET-CT scan immediately after gastrointestinal surgery demonstrated no brain metastasis. On admission to the ward, imaging of the brain showed right temporoparietal hematoma. In the ward, the patient deteriorated with impaired consciousness. Repeat brain imaging showed enlargement of the hematoma. He underwent right temporoparietal craniotomy for the removal of a hematoma, and tumor nodules adherent to the cortex was found. Pathology confirmed a metastatic DLBCL in the brain. Literature review showed that this was the first reported case of brain hemorrhage from metastatic lymphoma. Metastatic central nervous system lymphoma should be considered as a differential diagnosis in patients with a history of gastrointestinal lymphoma presenting with neurological symptoms.

Piloerection as the sole symptom of epilepsy: A case report and review of literature

Piloerection is an involuntary erection of body hairs that usually has physiological correlates such as cold or a strong emotional experience. Piloerection may also be a rare manifestation of seizure.Here, we report a case of 54-year-old man who experienced pilomotor seizures from temporal lobe epilepsy. The patient presented with sudden piloerection and no loss of consciousness many times a day. Magnetic resonance imagingof the brain showed threelesions in the right hemisphere, with the largest lesion in the right temporal lobe. A video-EEG showed an ictal discharge in the delta range with right temporal onset. Digital subtraction angiography excluded arteriovenous malformation. The lesion in the right temporal lobe was resected. Immunohistochemistry confirmed a cerebral cavernous malformation. There was no further seizure. A review of the published literature revealed that ictal piloerection as a lone manifestation is rare. Most cases of pilomotor seizure originate in the temporal lobe. Close to four fifth of the cases has a structural lesion. EEG was able to confirm the diagnosis of ictal piloerection in the majority of cases.

Rapid screening and identification of sesquiterpene lactones in Kudiezi injection based on high-performance liquid chromatography coupled with linear ion trap-orbitrap mass spectrometry / 中国天然药物

Sesquiterpene lactones are considered as the major active compounds in Kudiezi injection in virtue of their special structures and activities. Herein, an analytical method was developed for rapid screening and identification of sesquiterpene lactones in Kudiezi injection using high-performance liquid chromatography coupled with linear ion trap-orbitrap mass spectrometry (HPLC-LTQ-Orbitrap) in negative ion mode. First, two sesquiterpene lactone reference standards were analyzed to obtain their characteristic ESI-MS/MS fragmentation patterns. Second, based on extracted ion chromatography (EIC) data-mining method and characteristic fragmentation pathways analysis, sesquiterpene lactones in Kudiezi injection were rapidly screened and identified. Finally, an important parameter Clog P was adopted to discriminate the isomers of sesquiterpene lactones. As a result, 50 sesquiterpene lactones were characterized, including 9 sesquiterpene lactone aglycones, 39 sesquiterpene lactone glycosides, and 2 amino acid-sesquiterpene lactone conjugates. Among them, 13 compounds were tentatively identified as new compounds. The results demonstrated that the established method would be a rapid, effective analytical tool for screening and identification of sesquiterpene lactones in the complex system of natural medicines.

Metabolites of tanshinone Ⅰ and tanshinone ⅡA in rats / 中国中药杂志

An efficient method of ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap (UHPLC-LTQ-Orbitrap) mass spectrometer was established to elucidate the metabolites of tanshinone Ⅰ and tanshinone ⅡA in rats. Urine and plasma samples were collected after oral gavage. After processing biological sample by solid phase extraction, Waters ACQUITY HPLC BEH C₁₈ column (2.1 mm×100 mm, 1.7 μm) was used with 0.1% formic acid (A) - acetonitrile (B) solution as the mobile phase for gradient elution. The plasma, urine and the blank samples were then analyzed by ESI-LTQ-Orbitrap equipped with an ESI ion source under positive ion mode. On the basis of the accurate mass measurements, multiple mass spectra and comparison of data with published literature, a total of 26 metabolites were tentatively identified and characterized in the rat samples. Among them, 7 metabolites were derived from tanshinone Ⅰ through metabolic pathways of glucuronide conjugation, hydroxylation, reduction reaction, demethylation reaction, methylation, sulfate conjugation and their composite reactions. Nineteen metabolites were derived from tanshinone ⅡA through metabolic pathways of hydroxylation, reduction reaction, methylation, sulfate conjugation, glucuronidation, glucosylation and their complicated reactions. The results showed that the metabolism of tanshinone Ⅰ and tanshinone ⅡA in rats could be comprehensively clarified by using UHPLC-LTQ-Orbitrap mass spectrometer, providing material basis for the further research in terms of pharmacodynamics, toxicology, and secondary development of Chinese medicine.

Identification of metabolites of Danshensu in rats / 中国中药杂志

To identify the metabolites of Danshensu in plasma and urine in rats by using UHPLC-LTQ-Orbitrap method. After oral gavage of Danshensu CMC-Na suspension in SD rats, urine and plasma samples were collected and processed by solid phase extraction. ACQUITY UPLC BEH C₁₈ column (2.1 mm×100 mm, 1.7 μm) was utilized, with 0.1% formic acid (A)-acetonitrile (B) solution as the mobile phase for gradient elution. Negative electrospray ion mode based data-acquisition method was established to collect the mass spectrometry data of biological samples. As a result, Danshensu and 21 Danshensu Ⅰ phase and Ⅱ phase metabolites were finally identified according to the accurate mass measurements, mass fragmentation behaviors and comparing with the reference standards. The main metabolic pathways included dehydration, methylation, glucuronide conjugation, sulfate conjugation and their composite reactions. Consequently, our study expounded metabolites of Danshensu in rats based on UHPLC-LTQ-Orbitrap method and provided a reference for further researches on therapeutic material basis and mechanism of Danshensu.

Rapid screening and identification of sesquiterpene lactones in Kudiezi injection based on high-performance liquid chromatography coupled with linear ion trap-orbitrap mass spectrometry / 中国天然药物

Sesquiterpene lactones are considered as the major active compounds in Kudiezi injection in virtue of their special structures and activities. Herein, an analytical method was developed for rapid screening and identification of sesquiterpene lactones in Kudiezi injection using high-performance liquid chromatography coupled with linear ion trap-orbitrap mass spectrometry (HPLC-LTQ-Orbitrap) in negative ion mode. First, two sesquiterpene lactone reference standards were analyzed to obtain their characteristic ESI-MS/MS fragmentation patterns. Second, based on extracted ion chromatography (EIC) data-mining method and characteristic fragmentation pathways analysis, sesquiterpene lactones in Kudiezi injection were rapidly screened and identified. Finally, an important parameter Clog P was adopted to discriminate the isomers of sesquiterpene lactones. As a result, 50 sesquiterpene lactones were characterized, including 9 sesquiterpene lactone aglycones, 39 sesquiterpene lactone glycosides, and 2 amino acid-sesquiterpene lactone conjugates. Among them, 13 compounds were tentatively identified as new compounds. The results demonstrated that the established method would be a rapid, effective analytical tool for screening and identification of sesquiterpene lactones in the complex system of natural medicines.

Injury of human gastric epithelial GES-1 cells by MNNG and its effects on Wnt/β-catenin signaling pathway / 生理学报

The aim of this study was to investigate the mechanisms of mono-functional alkylating agent MNNG to damage human gastric epithelial GES-1 cells and roles of Wnt/β-catenin signaling pathway in the process. The GES-1 cells were treated with MNNG (2 × 10 mol/L) for 24 h. The morphological changes of the GES-1 cells were observed under inverted microscope 2 d after treatment. The cell viability was measured by MTT assay. The apoptosis and cell cycle distribution of the GES-1 cells were analyzed by flow cytometry. The mRNA expressions of β-catenin, GSK-3β, c-Met and MMP7 in the GES-1 cells were detected by qPCR. The protein expressions of β-catenin, GSK-3β, p-GSK-3β and c-Met were determined by Western blot. The results showed that MNNG induced the injury of GES-1 cells and changed the normal cell morphology to irregular long spindle shape. MNNG induced the apoptosis of GES-1 cells and blocked the cell cycle progression obviously. MNNG up-regulated the mRNA expressions of β-catenin, GSK-3β, c-Met and MMP7, and increased the protein expressions of β-catenin, GSK-3β and p-GSK-3β. These results suggest that the damage of GES-1 cells induced by MNNG may be related to the activation of Wnt/β-catenin signaling pathway, which will provide the basis for the study of cell model of gastric mucosal cell injury.

Neural stem cell-derived exosomes inhibit apoptosis of neurons induced by hypoxia neural cells / 中国病理生理杂志

AIM: To investigate whether neural stem cell-derived exosomes promote the viability and inhibit the apoptosis of neurons under cobalt chloride(CoCl2)-induced hypoxia in vitro.METHODS:The exosomes were isolated based on ultracentrifugation.The exosomal markers,ALG-2-interacting protein X(Alix)and tumor susceptibility gene 101 (TSG101)were identified by Western blot.The shape of exosomes was observed under transmission electron microscope (TEM).The size distributions of exosomes were analyzed by nanoparticle analysis(qNano).The neurons were exposed in CoCl2at different doses(200~600 μmol/L)for 24 h.The exosomes were co-cultured with the neurons pre-treated with CoCl2.The viability and apoptosis of the neurons were measured by CCK-8 assay and TUNEL method.RESULTS: The exosomes released from the neural stem cells expressed exosomal markers Alix and TSG 101.They also displayed a cup-shaped appearance observed under TEM and their sizes were(95.0 ±23.5)nm(n=370).The neuronal viability was sig-nificantly inhibited by CoCl 2in a dose-dependent manner(P<0.05).After treatment with exosomes,the viability of the neuron pre-treated with CoCl2was increased and the apoptotic rate was decreased(P<0.05).CONCLUSION: Neural stem cell-derived exosomes promote the viability and inhibit the apoptosis of rat neurons uneder hypoxia.

Effects of baicalein on proliferation and apoptosis of nasopharyngeal carcinoma CNE2 cells / 中草药

Objective To investigate the anti-proliferation effect of baicalein in nasopharyngeal carcinoma CNE2 cells and to clarify its mechanisms of cell cycle arrest and apoptosis. Methods The proliferative activity was detected by CCK-8 assay, and the changes of cell morphology was observed under fluorescence microscope. The rate of apoptosis and the cycle arrest were detected by flow cytometry, and caspase-3 activity was detected by caspase-3 activity kit. Expression of related genes (P53, P21, CDK2, and Caspase-3) at the RNA level were detected by using qRT-PCR. Results Baicalein had a significant inhibitory proliferation effect on CNE2 cells, which was the dose and time-dependent. After 48 h treatment with different concentrations of baicalein, the cells showed obvious apoptotic characteristics, with the increase of apoptotic cells, the activity of caspase-3 was increased, and S phase cells increase. Compared with the control group, the mRNA expression of P53, P21 and Caspase-3 increased, and mRNA expression of CDK2 decreased. Conclusion Baicalein could inhibit the proliferation of nasopharyngeal carcinoma CNE2 cells, and may induce apoptosis and cell cycle arrest in S phase by up regulating the expression of P53.

Delayed hemorrhage after microvascular decompression in a hemifacial spasm patient associated with neurosyphilis

Microvascular decompression is an effective treatment for hemifacial spasm. Hemorrhage is oneof the complications of microvascular decompression. However, delayed hemorrhage is very rare.Here, we report a case of ipsilateral cerebellar hemorrhage at day seven following microvasculardecompression. A 45-year-old woman presented with left HFS for the previous two years. Diagnostictesting demonstrated the presence of neurosyphilis. Brain magnetic resonance image was unremarkableon presentation. She received microvascular decompression and her hemifacial spasm completelyresolved after surgery. At day seven post-operatively, the patient presented with a sudden onsetheadache. Emergency computed tomography scan showed a cerebellar hemorrhage. A suboccipitalcraniotomy was performed and a cerebellar hematoma was evacuated. The delayed hemorrhage wasattributed to possible microaneurysm from syphilis.