Objective To study in vitro the effects of low-intensity pulsed ultrasound (LIPUS) on the proliferation and differentiation of cultured myoblasts,and to explore the cellular and molecular mechanisms behind any therapeutic effect of LIPUS.Methods Myoblasts were isolated from the skeletal muscles of mice and cultured in vitro.Treatment and control groups of proliferating and differentiating myoblasts were defined.The treatment groups were exposed to LIPUS at 1.5 MHz and a spatial and temporal average intensity of 30 mW/cm2,for 20 min daily,the proliferation group for 6 consecutive days and the differentiation group for 4 consecutive days.The cell proliferation kinetics of the proliferation group were analyzed using flow cytometry.The expression of myogenic regulation factor MyoD and heme oxygenase-1 (HO-1) in the proliferation group,and of myosin heavy chain (MHC) in the differentiation group were examined by immunofluorescent staining.Myoblast fusion indexes were analyzed.Results In the LIPUS treatment groups the proliferating myoblasts had a higher ratio of active cells in the G2 and S phases (19.30％ ±5.14％,37.00％ ±8.72％),compared with the controls (10.33％ ± 1.53％,25.00％ ±4.36％),and the proliferation index increased significantly.The expression of HO-1 was up-regulated,while MyoD staining was unchanged.During the induction of differentiation,the myoblasts of the treatment group fused into smaller myotubes and the myoblast fusion index (18.73％ ± 6.81％) was significantly lower than that of the control group (37.52％ ± 11.23％),while MHC expression did not change markedly.Conclusion LIPUS can promote myoblast proliferation while inhibiting their differentiation,but it does not affect the cells' myogenic properties.HO-1 may be involved in the regulation process.

Objective:To observe the effect of transdermal fentanyl combined with gabapentin for the treatment of malignant neu-ropathic pain (MNP). Methods:A total of 60 patients with MNP were randomly divided into two groups. A total of 30 cases in the con-trol group received transdermal fentanyl according to the dosages of opioid medicine that patients used. Such dosages were gradually in-creased until the pain relief visual analogue scale (VAS) fell below 3 or until the times of breakthrough pain became less than 3. For the combined group, gabapentin was co-administered with transdermal fentanyl, similar to the control group. Initially, 100 mg of gabapen-tin was administered thrice a day. This dosage was gradually increased until pain relief. However, gabapentin dosages were kept below 2,400 mg a day. VAS, quality of life (QOL), degree of pain relief, dosages of fentanyl and morphine, and side effects were evaluated be-fore treatment and at one, two, three, and four weeks after the treatment. Results:Both groups exhibited lower VAS after treatment (P<0.05), but the difference was observed to be more significant in the combined group than that in the control group (P<0.05). Both groups exhibited improved QOL (P<0.05), which was observed to be more significant in the combined group than in the control group (P<0.05). The effective rate was 96.7%in the combined group and 83.3%in the control group. The dosage of opioid medicine and the side effects in the combined group were less than those in the control group. Conclusion:Transdermal fentanyl combined with gabapen-tin is effective for the treatment of malignant neuropathic pain.

Objective To study the changes of hippocampal caspase-3 and PSD-95 expression levels in the mice exposed to ketamine 30 mg/(kg·d)for three months. Methods Forty C57BL/6 mice were randomly divided into two groups,and the chronic ketamine addiction model was established by giving mice a three month course of daily intraperitoneal injections of ketamine. Immunohistochemical study and Western blot-ting were applied to observe the expression of caspase-3 and PSD-95 protein. Results There were more expression of caspase-3 and less of PSD-95 in ketamine group as detected by immuohistochemistry. Western blotting results showed caspase-3 active fragment level significantly increased com-pared to saline group,but PSD-95 protein level was decreased. Conclusion The increased level of caspase-3 protein and reduced expression of PSD-95 are observed after long-term ketamine administration. These findings may provide an evidence for the neurotoxicity in mouse hippocampus of chronic ketamine addition as a recreational drug.

AIM:To explore the therapeutic effect of a novel Rho kinase inhibitor WAR 5 on the experimental autoimmune encephalomyelitis (EAE) and its possible mechanism.METHODS: Female C57BL/6 mice were randomly divided into EAE group and WAR5 group.EAE model was induced by the application of MOG 35-55 peptide.WAR5 was in-jected intraperitoneally every other day from post-immunization (PI) day 3 to PI day 27.The clinical score and body weight were recorded every other day .On PI day 28, the animals were sacrificed and spinal cords were obtained for HE and mye-lin staining .The splenocytes were isolated and the expression of CD 16/32 and CD206 were analyzed by flow cytometry . The protein extracts from the brains and spinal cords were collected for the measurement of inducible nitric oxide synthase ( iNOS) by Western blotting .RESULTS:The administration of WAR 5 delayed the onset of EAE and attenuated the clini-cal symptoms .The results of the pathological examination revealed that WAR 5 inhibited the infiltration of inflammatory cells and improved myelination in spinal cords , accompanied with the poralization of M 1 macrophages to M2 phenotype in the spleen.WAR5 inhibited the expression of iNOS in the central nervous system , especially in the spinal cords .CON-CLUSION:The therapeutic effect of WAR5 on EAE may be related to the shift of M1 macrophages to M2 phenotype and inhibition of inflammation in the central nerve system .

Objective To explore the clinic application value of ultrasonography examination of optic nerve sheath diameter(ONSD) in brain injury.Methods From July 2008-June 2011,90 cases of brain injured patients were chosen as experimental group including light (A group),medium (B group),and heavy (C group) brain injured patients according to the admission GCS score ;50 cases of conventional physical examination and 90 cases of volunteers 50 in neurosurgical outpatient were chosen as control group.The ONSD of both groups were measured 3 mm behind the globe through orbital using color sonographic with different time after admission.3 times measurements were carried out for every optic nerve sheath.All client's ONSD mean and standard deviation were calculated.In 0.5 h after color dopplar ultrasound examination,lumbar vertebra puncturing measured intracranial pressure in different groups.Results After admission (1d,3 d,7 d,14 d),the ONSD of A group was (4.54 ±0.32)mm,(4.42 ±0.30)mm,(4.44 ±0.32) m,and (4.43 ± 0.25) mm,respectively; The ONSD of B groups was (4.48 ± 0.28) mm,(4.52 ± 0.24) mm,(4.46 ±0.28)mm,and (4.38 ±0.22)mm,respectively; The ONSD of C group was (5.67 ±0.35)mm,(6.36 ± 0.42) mm,(5.65 ± 0.23) mm,and (4.76 ± 0.35) mm,respectively.After admission (1 d,3 d,7 d,14 d),the intracranial pressure (IP) of A group was (82 ± 11) mmH2O,(79 ± 12) mmH2O,(90 ±15) mmH2O,and (86 ± 14) mmH2O,respectively; The IP of B group was (78 ± 15) mmH2O,(85 ± 10)mmH2O,(78 ± 16) mmH2O,(80 ± 11) mmH2O,The IP of C group was (225 ± 26) mmH2 O,(288 ± 23)mmH2O,(256 ± 23) mmH2O,(122 ± 18) mmH2O,respectively.Group D had the ONSD average of (4.58± 0.41)mm and IP of (88 ± 10)mmH2O after eyeball 3-mm place.No difference was found between A and B,A and D,or B and D (P＞0.05) ; A difference was found between A and C,B and C,or D and C (t =12.24～24.67,P＜0.01).Conclusions The ONSD and IP in light medium brain injured patients had no change.In patients with severe brain injury,IP changed with the time after injury,the ONSD increased with the IP,the ultrasonography examination of ONSD with the important value in the diagnosis and treatment can respond the IP increase,which is a non-invasion,convenient,fast,and feasible method for evaluation of cranial high pressure.

Objective: To investigate the expression and clinical significance of long-chain non-coding RNA brain cytoplasmic RNA 1 (BCYRN1) in serum of patients with non-small cell lung cancer. Methods: 74 patients with non-small cell lung cancer (NSCLC) diagnosed and treated in our hospital were selected as the experimental group, and 50 healthy subjects were selected as the control group. The expression of BCYRN1 in serum of NSCLC and control group was detected by real-time quantitative polymerase chain reaction (qRT-PCR). The patients′ clinical pathology data were collected and followed up. To analyze the relationship between the expression of serum BCYRN1 and clinicopathological parameters of NSCLC patients, and the relationship between BCYRN1 expression in the diagnosis and prediction of NSCLC prognosis. Results: The relative expression of BCYRN1 in serum of NSCLC patients was 2.84±0.95, which was significantly higher than that of healthy controls (1.16±0.50) (P<0.05). The level of serum BCYRN1 in patients with NSCLC with tumor size >3 cm was significantly higher than that in patients with tumor size ≤3 cm (P<0.05). The level of BCYRN1 in patients with tumor node metastasis (TNM) Ⅲ+ Ⅳ was significantly higher than that in patients with stage Ⅰ+ Ⅱ (P<0.05). Patients with low differentiation and positive lymph node metastasis had higher serum BCYRN1 than those with moderate to high grade and lymph node metastasis (P<0.05). The receiver operating characteristic (ROC) curve analysis showed that the area under the curve was 0.847 (95% CI: 0.772-0.922, P=0.000), and the sensitivity and specificity were 68.9% and 88.0%, respectively. The median progression free survival (PFS) (15 vs 21 months) and overall survival (OS) (19 vs 28 months) of patients with high and low expression of BCYRN1 were statistically significant (P<0.05). High expression of BCYRN1 was an independent risk factor for overall survival in patients with NSCLC. Conclusions BCYRN1 is highly expressed in the serum of NSCLC patients, and high expression of BCYRN1 is closely related to the poor prognosis of patients with NSCLC. It can be used as a novel biomarker and diagnostic target for NSCLC.

Objective:To detect the expression of long non-coding RNA (LncRNA) ARAP1-AS1 in pancreatic cancer, and to preliminarily explore its effects on the biological behaviors of proliferation, apoptosis, migration and invasion of pancreatic cancer cell.Methods:The pancreatic cancer tissue specimens and corresponding paracancerous tissue specimens of 25 patients were collected, and the expression of ARAP1-AS1 was detected by qPCR. Human pancreatic cancer cell line PANC-1 was cultured in vitro and divided into control group, siRNA-control group (transfected with siRNA control sequence), knockout group (transfected with ARAP1-AS1 siRNA), pcDNA3.1-control group (transfected with pcDNA3.1) and overexpression group (transfected with pcDNA3.1-ARAP1-AS1), qPCR method was used to detect the transfection efficiency, CCK-8 method was used to detect the cell proliferation ability, flow cytometry was used to detect the cell apoptosis, scratch test was used to detect the cell migration ability, Transwell method was used to detect the cell invasion ability, Western blot (WB) method was used to detect the expression of proliferating cell nuclear antigen (PCNA), B lymphoma-2 protein (Bcl-2), Bcl-2 related X protein (Bax), matrix metalloproteinase-9 (MMP-9) proteins.Results:The expression level of ARAP1-AS1 in pancreatic cancer tissues was significantly higher than that in adjacent tissues (2.26±0.13 vs 1.00±0.00) ( P<0.05). Compared with the siRNA-control group, the ARAP1-AS1 level (1.01±0.02 vs 0.29±0.03), PCNA, Bcl-2, MMP-9 protein levels, cell OD value (0.57±0.05 vs 0.23±0.03), scratch healing rate (78.53±7.02 vs 48.60±5.26), and number of invasions (229.63±22.59 vs 104.25±15.04) in PANC-1 cells of the knockout group were significantly reduced ( P<0.05), the Bax protein level and the apoptosis rate (4.52±0.42 vs 32.40±1.84) were significantly increased ( P<0.05). Compared with the pcDNA3.1-control group, the ARAP1-AS1 level (1.02±0.03 vs 2.06±0.08), PCNA, Bcl-2, MMP-9 protein levels, cell OD value (0.57±0.05 vs 0.90±0.08), scratch healing rate (77.65±6.67 vs 91.22±7.34), and number of invasions (225.34±19.65 vs 327.50±25.40) in PANC-1 cells of the overexpression group were significantly increased ( P<0.05), the Bax protein level and the apoptosis rate (4.58±0.48 vs 2.29±0.24) were significantly reduced ( P<0.05) . Conclusion:LncRNA ARAP1-AS1 is highly expressed in pancreatic cancer, which can promote the proliferation, migration and invasion of pancreatic cancer cells PANC-1, and reduce cell apoptosis.

Objective To investigate the association between genetic variants in microRNA biosynthesis genes and the risk of head and neck squamous cell carcinoma (HNSCC).Methods A case-control study was conducted with 576 HNSCC patients and 1 552 healthy controls matched by factors as age-(± 5 years) and sex.Eight potentially functional single nucleotide polymorphism loci in microRNA biosynthesis genes (DICER1,GEMIN3,and PIWIL1) were genotyped using the Illumina Infinium BeadChip platform.Univariate and multivariate logistic regression models were performed to assess the association between genotypes and HNSCC risk.Results The allele frequencies of rs1106042 (G＞A) in PIWIL1 were significantly different between the cases and controls (P=0.011).After controlling for factors as age,sex,smoking and alcohol intake,the A allele of rs 1106042 showed a decreased risk of HNSCC (additive model:adjusted OR=0.73,95％CI:0.57-0.93,P=0.011).Results from the stratification analysis by age,sex,smoking,alcohol intake and tumor sites showed that the effect of rs1106042 A allele on HNSCC risk was significant in older age groups (≥60),females,nonsmokers,non-alcohol drinkers,and subjects with oral cavity cancer (P＜0.05).Conclusion Potentially,functional single nucleotide polymorphism in PIWIL1 might modify the risk of HNSCC in China.

Objective:To analyze the factors affecting the disappearance time of airway necrosis and repair time of airway scar stenosis in patients with ulceration necrosis tracheobronchial tuberculosis (TBTB Ⅱ) after standardized chemotherapy and bronchoscopic intervention.Methods:The clinical data of 222 TBTB Ⅱ patients admitted to Hunan Chest Hospital from January 2015 to December 2018 were collected, bronchoscopic interventional treatment was performed on time. The texture, blockage of lumen, granulation proliferation, airway stenosis of TBTB patients before treatment, the disappearance time of airway dead objects, scar repair time and stenosis degree after treatment were followed up. The disappearance time of airway necrosis and repair time of airway scar stenosis and its influencing factors were recorded and analyzed.Results:In 222 patients, 508 ulceration necrosis airway lesions were found under bronchoscopy, with a median of 2(1-6); 170(76.6%) cases of airway lesions had different degrees of stenosis before treatment. 79(35.6%) patients had tough necrosis, and 86(38.7%) patients had necrosis blocking the lumen; 132(59.5%) patients had granulomatosis. The disappearance time of airway necrosis after treatment was 1 to 32 weeks, and M( Q1, Q3) was 6(3, 9) weeks; the repair time of airway scar stenosis was 2 to 73 weeks, and M( Q1, Q3) was 14(10, 19) weeks; after treatment, there were 90.5%(201/222) patients with different degrees of scarring in the airways. Cox multiple analysis showed that the risk factor for the disappearance time of airway necrosis was tough tough necrosis ( HR=1.52, 95% CI: 1.10-2.10); the risk factor for the repair time of airway scar stenosis was the disappearance time of airway necrosis 6-9 weeks ( HR=2.73, 95% CI: 1.84-4.05). Conclusions:90.5% of patients with type Ⅱ TBTB developed airway scar stenosis after treatment. The median time for the disappearance of airway necrosis was 6 weeks, and the median time for the repair time of airway scar stenosis was 14 weeks. In the interventional process, attention should be paid to the removal of tough necrosis and the efficiency of necrosis removal to reduce the risk of airway scar stenosis.

BACKGROUND:Cerebral ischemic stroke is one of the main fatal and disabling diseases in the clinic,but only a few patients benefit from vascular recanalization in time,so it is urgent to explore new and effective therapy.As one of the critical pathological changes of ischemic stroke,the glial scar formed mainly by astrocytes is one major cause that hinders axonal regeneration and neurological recovery at the late stage of stroke. OBJECTIVE:To elucidate the pathological process and crucial signal regulatory mechanism of astrocytes in the formation of glial scar after ischemic stroke,as well as the potential therapeutic targets,to provide a theoretical reference for intervening astrocytic scar formation against ischemic stroke effectively,and novel strategies for promoting post-stroke rehabilitation. METHODS:The relevant articles published in CNKI,PubMed and Web of Science databases from 2010 to 2022 were retrieved.The search terms were"Ischemic stroke,Brain ischemi*,Cerebral ischemi*,Astrocyt*,Astroglia*,Glial scar,Gliosis,Astrogliosis"in Chinese and English.Finally,78 articles were included after screening and summarized. RESULTS AND CONCLUSION:(1)Astrocytes play an important role in the maintenance of central nervous system homeostasis.After ischemic stroke,astrocytes change from a resting state to an active state.According to the different severities of cerebral ischemic injury,astrocyte activation changes dynamically from swelling and proliferation to glial scar formation.(2)Mature astrocytes are stimulated to restart the cell cycle,then proliferate and migrate to lesions,which is the main source of the glial scar.Neural stem cells in the subventricular zone,neuron-glial antigen 2 precursor cells and ependymal precursor cells in the brain parenchyma can also differentiate into astrocytes.Endothelin-1,aquaporin 4,ciliary neurotrophic factor and connexins are involved in this process.In addition,chondroitin sulfate proteoglycan,as the main component of the extracellular matrix,forms the dense glial scar barrier with proliferated astrocytes,which hinders the polarization and extension of axons.(3)Activation or inhibition of crucial signal molecules involved in astrocyte activation,proliferation,migration and pro-inflammation functions regulate the glial scar formation.Transforming growth factor beta 1/Smad and Janus kinase/signal transducer and activator of transcription 3 are classical pathways related to astrogliosis,while receptor-interacting protein 1 kinase and glycogen synthase kinase 3β are significant molecules regulating the inflammatory response.However,there are relatively few studies on Smad ubiquitination regulatory factor 2 and Interleukin-17 and their downstream signaling pathways in glial scar formation,which are worthy of further exploration.(4)Drugs targeting astrogliosis-related signaling pathways,cell proliferation regulatory proteins and inflammatory factors effectively inhibit the formation of glial scar after cerebral ischemic stroke.Among them,the role of commonly used clinical drugs such as melatonin and valproic acid in regulating glial scar formation has been verified,which makes it possible to use drugs that inhibit glial scar formation to promote the recovery of neurological function in patients with stroke.(5)Considering the protective effects of glial scar in the acute phase,how to choose the appropriate intervention chance of drugs to maintain the protective effect of the glial scar while promoting nerve regeneration and repair in the local microenvironment is the direction of future efforts.