Objective To investigate the development of rehabilitation medicine, the knowledge, ability and quality requirements of stu-dents in different professional positions, and the demand for rehabilitation industry personnel in Hunan, China, and to discuss the strategies for improving the quality of vocational education. Methods 87 directors of personnel management or rehabilitation department in all levels of hospitals and 100 rehabilitation therapists were included from May to Semptember 2014. The rehabilitation medical service, education background of employees, professional positions, and the demand for rehabilitation knowledge, ability, and industry personnel were sur-veyed. Results and Conclusion Rehabilitation medical service was not sufficiently carried out for insufficient number of employees, unbal-ance structure and other deficiencies. Rehabilitation personnel training schools and enterprises need to strengthen the depth of integration, and improve the professional quality and clinical ability.

Objective To determine the relationship between early postoperative delirium (EPD) and prognosis in patients undergoing non-cardiac surgery. Methods This was a prospective cohort study consisted of 698 patients admitted to postanesthesia care unit, undergoing non-cardiac surgery under general anesthesia, between June and December 2009. The risk factors affecting prognosis were collected. All the patients were assessed for the development of delirium by experienced research staff using Confusion Assessment Method for Intensive Care Unit. The patients were divided into 2 groups according to the occurrence of EPD: EPD group and no EPD (NEPD) group. The postoperative hospital length of stay was made as a major prognostic indicator. Cox proportional hazard regression model was used to analyze the risk factors affecting prognosis. Results Of the 698 patients, 197 (28.2%) developed EPD. The postoperative hospital length of stay was prolonged in group EPD compared with group NEPD. The Cox proportional hazard regression model analysis indicated that EPD was an independent risk factor affecting prognosis. Conclusion EPD is an independent risk factor affecting prognosis in patients undergoing non-cardiac surgery.

[ ABSTRACT] AIM:To construct a lentiviral vector for stable delivery of the ER-α36 gene and to detect its effect on SGC7901 cell growth.METHODS: The efficient RNAi targeting sequences identified for the ER-α36 gene were screened.The Oligo DNA was synthesized with target sequences and annealed to form double-stranded DNA.Then it was digested by Xho I and EcoR I and connected with GV307 vector to produce LV-ER-α36-RNAi lentiviral vector.PCR was used to screen the positive clones and sequence.The LV-ER-α36-RNAi, pHelper 1.0 and pHelper 2.0 plasmids were co-transfected into 293T cells for producing lentiviral vector and infecting SGC7901 cell line.Fluorescence microscopy, real-time PCR and Western blotting were used to detect the transfection efficiency and gene silencing effect.17β-estrodial at concentration of 1 ×10 -10 mol/L was used to stimulate the recombinant cell line, and the action on the growth of gastric cancer cells and the expression of Src, ERK1/2 and cyclin D1 were determined.RESULTS: DNA sequencing analysis confirmed the identity of recombinant shRNA expression vectors.Immunofluorescence assay demonstrated that transfection efficiency was above 80%.Transfection of LV-ER-α36-RNAi significantly knocked down the expression of ER-α36 at mR-NA and protein levels with tetracycline ( TeT) simulating as revealed by real-time PCR and Western blotting.Compared with control group, the growth of the recombinant cell line declined and the expression of Src, ERK1/2 and cyclin D1 and the activation of Src decreased (P<0.05).CONCLUSION: Lentiviral vectors that silence ER-α36 expression are con-structed successfully and can be used to study the role of ER-α36 in gastric cancer.The ER-α36 is related with many kinds of cancer cell growth, including gastric cancer cells.

Objective:To investigate the potential mechanism of Shenfu injection in regulating stress response via the neuro-endocrine-immune system by network pharmacology and molecular docking. Methods:The main active ingredients and related targets of Shenfu injection were screened using the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform.PharmMapper, Swiss Target Prediction platform and Uniprot database were used to predict the target and unify the gene names.GeneCards, OMIM, TTD, CTD, Drugbank, Disgenet and Pharmgkb databases were searched to screen the related targets regulated by stress responses.Venny 2.1 tool was used to obtain the potential effect targets of the intersection between Shenfu injection and stress response regulation, and the STRING database was imported to construct the interaction PPI network and screen the key targets.Potential effect targets were uploaded to Metascape database online analysis for study on the mechanism through GO and KEGG enrichment analysis.Autoduck and Pymol were used for molecular docking and visualization.Results:Forty-three main active ingredients and 257 related targets for Shenfu injection were obtained by component screening and target prediction.A total of 4 811 targets related to stress response regulation were retrieved from the database, 188 potential effect targets were obtained by intersection with Shenfu injection component-related targets, and 14 key targets were obtained by PPI network screening.Eighteen samples were screened by GO enrichment analysis, which mainly involved the circulatory system and humoral regulation, responses to external stimuli and trauma, MAPK cascade reaction, postsynaptic membrane, receptor complex and ion channel complex and neurotransmitter receptor activity, etc.KEGG enrichment analysis showed 20 highly correlated pathways, mainly covering neuroactive ligand-receptor interaction, calcium signaling pathway, adrenergic signaling, steroid hormone biosynthesis, IL-17, TNF, MAPK, cGMP-PKG, PI3K-Akt, NF-κB, Toll-like receptor signaling pathway and cell apoptosis, etc.The results of molecular docking indicated that the main active components had good binding force with the key target.Conclusions:The components of Shenfu injection such as kaempferol, β-sitosterol, Demethyldelavaine, Stigmasterol, ginsenoside, Carnosifloside, hypaconitine may act on targets such as AKT1, TNF, IL1B, PTGS2, HSP90AA1, MAPK1, NFKBIA, NR3C1 and ADRB2 and regulate the stress response through the mechanisms such as regulation of the functional state of the neuro-endocrino-immune system, inhibition of inflammatory responses, anti-oxidative stress and reduction of cell apoptosis.

Objective To evaluate the effect of acupuncture(manual acupuncture,electroacu-puncture)on prevention and treatment of nausea and vomiting after general anesthesia.Methods By searching CNKI,VIP,China Biomedical Literature Database,Wanfang Medical,Embase,PubMed,Co-chrane Clinical Trial Registry,randomized controlled trial(RCT)on acupuncture combined with antiemetics for the prevention and treatment of nausea and vomiting after general anesthesia were performed.The retrieval period was from the establishment of the database to September 2022.RevMan 5.3 was used for statistical analysis.Results Fifteen studies involving 1 493 patients were included.There were 741 patients in the antiemetic group and 752 patients in the acupuncture combined antiemetic group.The incidence of postoperative nausea in the acupuncture combined with antiemetics group was significantly lower than that in the simple antiemetics group(OR=0.43,95％CI 0.34-0.54,P＜0.001).The incidence of postopera-tive vomiting in the acupuncture combined with antiemetics group was significantly lower than that in the simple antiemetics group(OR=0.55,95％CI 0.45-0.66,P＜0.001).Conclusion The incidence of postoperative nausea and vomiting was lower in patients treated with acupuncture(manual acupuncture,electroacupuncture)combined with antiemetics than with antiemetics alone.

Objective To explore the effect of calcitonin gene-related peptide (CGRP) on expressions of aquaporin (AQP)1 and AQP 5 in young rats with acute lung injury (ALI) caused by lipopolysaccharide (LPS).Methods Eighty-four young rats were randomly divided into control group,ALI model group and CGRP group.The rats in ALI model group were given intraperitoneal injection of LPS (5 mg/kg)for 2,6,12,24 hours;while the rats in CGRP group were given intraperitoneal CGRP (1 mg/kg) after 1 h injection of LPS.At 2 h,6 h,12 h and 24 h,all rats were sacrificed and lung tissues were obtained.The histopathological changes in lung tissues were evaluated by adopting hematoxylin-eosin staining,and wet/dry(W/D) was measured.The mRNA and protein levels of AQP1 and AQP5 in lung tissues were detected by adopting fluorogenic quantitative PCR (qPCR) and Western blot.Results Pathological stain showed that rats in control group had a normal lung tissue structure,and LPS made lung tissue edema,narrowing the alveolar cavity and inflammatory cell infiltration.CGRP attenuated the effect of LPS on rat's lung.The W/D ratio of lung tissue was significantly higher than that in the control group,and CGRP reduced the W/D ratio of lung tissue.qPCR showed that the mRNA levels of AQP1 and AQP5 from rats in ALI group (0.009 ±0.001 and 0.055 ±0.006)decreased compared with those in the control group (0.035±0.002 and 0.167 ±0.006) and CGRP group (0.024 ± 0.002 and 0.134 ± 0.012) (all P ＜ 0.001).Western blot results showed after 24 h injection of LPS,both AQP1 and AQP5 levels from ALI group (0.397 ± 0.041 and 0.215 ± 0.029) were significantly lower than those in the control group (0.850 ± 0.020 and 0.741 ± 0.032) (all P ＜ 0.001),and their levels in CGRP group (0.593-± 0.065 and 0.461 ± 0.039) were also lower than those in the control group,but higher than those in ALI group (all P ＜ 0.001).Conclusion CGRP can enhance AQP1 and AQP5 levels and reduce pulmonary edema,and it has a protective effect on rats with acute lung injury.

Objective To construct engineered corneal epithelium from embryonic stem cells (ESCs) using Rock inhibitor combined with hypoxia-normoxia culture condition. Methods Human ESC line H1 was induced to differentiate into epithelial-like cells by addition of retinoic acid (RA) and bone morphogenetic protein 4(BMP4) in the differentiation medium under the adherent culture condition. The ESCs derived epithelial-like cells were expanded in the mixed medium of SHEM and KSFM with the mixture ratio of 1 : 2 with or without Rock inhibitor Y27632. The H1 derived epithelial-like cells were seeded on the denuded ammonic membrane to construct engineered corneal epithelium under hypoxia,normoxia and hypoxia-normoxia culture conditions,respectively. The inducted effect of ESCs into epithelial-like cells,the expansion ability of the epithelial-like cells and the characteristics of the constructed engineered corneal epithelium were evaluated by morphological observation, real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and immunofluorescence technology. Results Compared with the control group,the relative expressions of ESCs marker Oct4 mRNA, Notch signaling pathway related factors Notch1 and Jagged1 mRNA,and Wnt signaling pathways related factors c-myc and Cyclin D1 mRNA were significantly reduced, and the relative expressions of cutaneous ectoderm markers p63 and K18 mRNA were significantly increased at day 8 after induction in the induced group,with significant differences between them (t =14.63,20.15,93.50,11.60, 19.30,18.44,22.63;all at P<0.05). Compared with the without Y27632 group,the relative expressions of p63 and K14 mRNA,Notch signal pathway receptor Notch1 and Jagged1 mRNA were significantly increased,and Wnt signaling pathways downstream targeted gene c-myc and CylinD1 mRNA were significantly decreased at day 8 after induction in the Y27632 group,with significant differences between them (t =20.29,59.22,2.90,39.59,5.32,10.14;all at P<0.05),and the relative expression of K18 mRNA in the two groups was not significantly changed(t=1.38,P>0.05). The ESCs derived epithelium and constructed under hypoxia-normoxia culture condition showed more obvious stratification and tighter cell arrangement in comparison with those cells cultured in consistent hypoxia culture condition or normoxia culture condition. Epithelial markers Pan-CK and K18 as well as epithelial progenitor cell markers p63 and K14 expressed in the whole cell layers of the ESCs derived epithelium constructed under hypoxia-normoxia culture condition. Conclusions The addition of Y27632 enhances the proliferation ability of H1 derived epithelial cells and actives Notch signaling pathway and inhibits Wnt signaling pathway. The culture and construction in the expansion medium with Y27632 under the hypoxia-normoxia culture condition can promote the stratification of H1 derived engineered corneal epithelium.

BACKGROUND:In recent years,increasing studies have focused on the abnormal proliferation and differentiation of acinous cells in the meibomian gland,suggesting that this process is closely related to the occurrence and development of dry eye.Structural and functional abnormalities such as blockage of the lumen of the meibomian gland and atrophy of the glands can cause or exacerbate dry eye.Therefore,the study of changes in the meibomian glands in dry eyes is important for understanding the pathogenesis of dry eyes in depth and finding new targets for the treatment and prevention of dry eyes. OBJECTIVE:To investigate the changes of the meibomian gland in a mouse model of aqueous deficient dry eyes. METHODS:Thirty-two female C57/B6 mice at 6-8 weeks were selected and randomly divided into experimental and control groups with 16 mice in each group.The mice in the experimental group were constructed by removing both the extra-orbital and intra-orbital lacrimal glands,while those in the control group were not treated.After 2 weeks of normal feeding,the corneal changes of both groups were observed under a slit lamp,and the tear secretion of both groups was measured.The meibomian glands of the two groups of mice were removed after decapitation.The changes in the gross morphology of the meibomian glands were observed and the meibomian glands were made into frozen sections.Hematoxylin-eosin staining was used to observe the structure of the meibomian glands,oil red staining was used to evaluate the function of the meibomian glands,and immunofluorescence staining and RT-qPCR were used to observe the expression of cytokeratin 14,Ki67 and abnormally differentiated small proline-rich protein 1B in the meibomian glands of mice. RESULTS AND CONCLUSION:Two weeks after modeling,lamellar defects were seen in the corneas of the experimental mice,and neovascularization of the limbal corneal was generated and invaded the central cornea.(2)Tear secretion volume was significantly reduced in the experimental group compared with the control group(P<0.05).Microscopic findings showed that the ducts of the meibomian glands in the experimental group were interrupted and atrophied,and their arrangement was disorganized.Hematoxylin-eosin staining results showed a significant increase in lipid vacuoles in the meibomian glands of the experimental mice compared with the control group.Lipid deposition was seen in oil red staining in the experimental group.Immunofluorescence and RT-qPCR results showed a significant increase in the expression of cytokeratin 14,Ki67 and small proline-rich protein 1B in the meibomian glands of mice in the experimental group compared with the control group(P<0.05).To conclude,aqueous deficient dry eye can lead to compensatory hypertrophy,increased proliferation,and abnormal lipid metabolism in the meibomian gland,as well as abnormal differentiation of the meibomian gland.

Objective To observe the effect of Shenfu injection on lung injury caused by hemor-rhagic shock(HS)in rats and explore the related potential mechanism.Methods Thirty-six SPF healthy male SD rats,aged 16-17 weeks,weighing 400-600 g,were randomly divided into three groups:sham op-eration group(group SH),HS group(group HS),and Shenfu injection group(group SF),12 rats in each group.In group SH,only the right femoral vein and femoral artery were separated after anesthesia,and ve-nous catheterization was not performed.HS model was established in groups SF and HS.In group HS,liquid resuscitation was performed through an intravenous catheter,and the resuscitation fluid consisted of the auto-blood lost and the compound sodium chloride injection equivalent to 1.5 times the blood loss and 10 ml/kg normal saline.In group SF,the resuscitation fluid consisted of the lost autoblood and the compound sodium chloride injection equivalent to 1.5 times the blood loss and Shenfu injection 10 ml/kg.The whole perfusion time was about 60 minutes.Six rats in the three groups were randomly anesthetized 24 and 48 hours after op-eration.The wet/dry weight ratio(W/D)of lung tissues was detected.The concentrations of interleukin-6(IL-6),IL-17,IL-10,and transforming growth factor-β(TGF-β)were detected by ELISA,the mRNA ex-pression of retinoic acid-related orphan nuclear receptor γt(RORγt),transcription factor forkhead box pro-tein 3(Foxp3),and hypoxia-inducible factor-1α(HIF-1α)in lung tissues were detected by PCR.The pro-tein contents of RORγt,Foxp3,HIF-1α,aquaporin 1(AQP1),and AQP5 in lung tissue were detected by Western blot.Pathological changesunder HE staining light microscope and lung injury scores were observed.Results Compared with 24 hours after operation,W/D,the concentrations of IL-6 and IL-17,mRNA ex-pression and protein content of RORγt and HIF-1α,and lung injury score were significantly decreased(P＜0.05),the concentrations of IL-10,and TGF-β,Foxp3 mRNA expression and protein content,and AQP1 protein content were significantly increased in group SF 48 hours after operation(P＜0.05).Compared with group SH,W/D,the concentrations of IL-6,IL-17,IL-10,and TGF-β,mRNA expression and protein content of RORγt,Foxp 3,and HIF-1α,and lung injury score were significantly increased(P＜0.05),AQP1 and AQP5 protein contents were significantly decreased in groups HS and SF 24 and 48 hours after operation(P＜0.05),and alveolar structure was damaged under light microscope and alveolar interstitium was filled with a large amount of edematous fluid,during which a large number of inflammatory cells infiltra-ted.Compared with group HS,W/D,the concentrations of IL-6 and IL-17,mRNA expression and protein content of RORγt and HIF-1α,and lung injury score were significantly decreased(P＜0.05),the concen-trations of IL-10 and TGF-β,Foxp3 mRNA expression and protein content,AQP1 and AQP5 protein con-tents were significantly increased in group SF 24 and 48 hours after surgery(P＜0.05),and the alveolar structure was improved under light microscope,and edema was reduced,and the number of inflammatory cells was reduced.Conclusion Shenfu injection can regulate the balance between pro-inflammatory factors IL-6 and IL-17,and anti-inflammatory factors IL-10 and TGF-β,increase the protein content of AQP1 and AQP5 in lung tissue,and decrease the W/D and injury score in lung tissue,thus alleviating lung injury in HS rats.The mechanism may be related to the regulation of HIF-1α-RORγt/Foxp3 balance.