BACKGROUND:Human skin-derived precursors can be cultured for a long term in vitro, and differentiated into neurons, glial cel s, smooth muscle cel s, Schwann cel s and cel s with peripheral neurons phenotype.
OBJECTIVE:To investigate the culture conditions and multiple differentiation capacity of multipotential stem cel s from human skin, especial y the potentials of differentiating into neurons and osteoblasts.
METHODS:Human skin-derived precursor cel s were cultured with trypsin digestion method, and identified with immunocytochemistry. Cel s at passages 3-4 were induced to differentiate into neurons and osteoblasts, and underwent von Kossa staining protocol for calcium, chondrocyte induction, toluidine blue staining, immunohistochemical staining and Sudan black staining. The expression of nestin, vimentin,βIII-tubulin, S100 and col agen II in the human skin-derived precursors was detected.
RESULTS AND CONCLUSION:The human skin-derived precursor cel s cultured with trypsin digestion method could proliferate and form suspending spheres, and nestin positive cel s were detected at any time point of the culture. Al the cultured cel s expressed vimentin, and some adherent cel s expressedβIII-tubulin. Human skin-derived precursor cel s were induced with Salvia miltiorrhiza to differentiate into neuron-like cel s, and expressed marker of nerve cel s. Skin-derived precursors could be induced to differentiate into osteoblasts and von Kossa staining displayed black calcified nodules in the culture dish. Skin-derived precursors could also be induced to differentiate into chondrocytes, and toluidine blue staining was strongly positive, and some cel s expressed col agen II, which suggested that, the differentiated cel s contained chondrocytes. Experimental findings indicate that, skin contains multipotential stem cel s that are capable of differentiating into osteoblasts, chondrocytes, Schwann cel s and oligodendrocytes.

BACKGROUND:There are myoblasts in human embryonic skeletal muscle. It remains poorly understand whether myoblasts in vitro can form myotube and what are the corresponding markers for identifying myoblasts and myotubes. OBJECTIVE:To investigate whether in vitro cultured myoblasts from human embryonic skeletal muscle can form myotube and whether they can express neural markers. METHODS:Human embryonic muscle-derived myoblasts were cultured in serum-containing medium. When the primary culture was established, cultured cel s were identified with immunocytochemistry for neural markers, such asβ-tubulin markers (desmin, myogenin, smooth muscle actin and myosin). RESULTS AND CONCLUSION:A population of myoblasts could migrate from human embryonic muscle tissues. They could express the markers for skeletal muscle such as desmin and myogenin, and they could express neuron specific enolase, nestin and neurofilament 200. They could form myotubes in vitro, and myotubes expressedβⅢ-tubulin, neurofilament 200 and glial fibril ary acidic protein. The data support the hypothesis that myoblasts from human embryonic muscle express neural markers and muscle markers, and cultured myoblasts and myotubes expressed neuron specific enolase,β-tubulin Ⅲ, nestin, neurofilament 200 and glial fibrillary acidic protein. This indicates that these markers could not be used for cel identification of trans-differentiation study from muscle origin to nervous system.

Objective To find a ideal method for inguinal herniorrhaphy. Methods One hundred patients with inguinal hernia were randomized to receive respectively PHS and Mesh Plug & Patch tension-free herniorrhaphy. Results In PHS group, the average operative time was 35 min. The postoperative complication rate was 2% (1/50), the average hospital stay was 2 days. Follow-up of 6～10 months has found no recurrence. In Plug group, the average operative time was 40 min,complication rate was 8% (4/50), the average hospital stay were 3 days. Postoperatively, 6 patients complained of temporary foreign body feeling. Follow-up of 6～10 months has found no recurrence in either group. Conclusions Both PHS and Mesh Plug & Patch are ideal materials for herniorrhaphy. PHS is especially suitable for lean inguinal hernia patients with large defect on transversalis fascia. Plug & Patch has an advantage in fat patients with small defect on transversalis fascia.

BACKGROUND:At present,the most frequently agents used for neural induction of bone marrow stromal cells(BMSCs)in vitro are anti-oxidants,such as beta-mercaptoethanol and all trans-retinoids.The majorities of induction from BMSCs are neuron-like cells in these protocols;however,whether it has neuronal function or not should be further studied.OBJECTIVE:TO investigate the differentiated characteristics of inducing human BMSCs into neural cells in serum-free medium.DESIGN:Observational study.SETTING:Chaozhou Central Hospital.MATERIALS:The experiment was carried out in the Chaozhou Central Hospital from April 2004 to December 2005.Adult bone marrows were derived from femoral and tibial bone marrow of three patients with fracture.All patients provided the confirmed consent and were approved by the Ethics Committee of Chaozhou Central Hospital.DMEMIF12 medium(1:1),fetal bovine serum (FBS), glutamine, N2 supplements and B27 Supplements were from GIBCO/BRL Company;recombinant basic fibroblast growth factor(bFGF)and recombinant epidermal growth factor(EGF)from Sigma Company;monoclonal antibody for vimentin(1:100),monoclonal antibody for myelin basic protein(MBP) (1:100),monoclonal antibody for S1 00(1:1 00),monoclonal antibody for neuron specific enolase(NSE)(1:1 00),and monoclonal antibody for neurofilament 200(NF200)(1:1 00)from Beijing Zhongshan Company;monoclonal antibody for glial fibrillary acidic protein (GFAP)(1:200)and polyclonal antibody for nestin(1:100)from Boster Company(Wuhan);mouse monoclonal antibody for beta-tubulin 3(1:1 000)from Sigma Company;SP-9000 kits and quick AEC from Beijing Zhongshan Company; culture dishes and flasks from Coming Company.METHODS:BMSCs from human bone marrow were cultured in serum-containing medium.When the primary culture was established, BMSCs were transferred into serum-free medium containing N2 or B27 supplement with 20 μg/L basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF),and cells were cultured in an incubator containing C02of 0.05 volume fraction at 37℃. Morphological changes of BMSCs in serum-free medium were observed under phase contrast microscope. And two days after culture. Expression of relative markers of BMSCs was detected withimmunocytochemistry.MAIN OUTCOME MEASURES:Morphological changes of BMSCs and expression of relative markers of nerve cells.RESULTS:A population of BMSCs could be isolated from adult human bone marrow,and they were processed to obtain a fibroblast-like population and were expanded as undifferentiated cells in culture for more than 10 passages.indicating their proliferative capacity.They could form spheroid state when they were sub-cultured in serum-free media supplemented with bFGF and EGF.these cells could express the markers for neural stem cells such as vimentin and nestin;they could expressed neuron specific enolase(NSE),beta-tubulin 3,TrkC and neurofilament 200(NF200)when they were plated on dishes with serum-containing medium; some cells exhibited the phenotypes for astrocytes.expressing gilal fibrillary acidic protein(GFAP)and S100 protein.CONCLUSION:The morphology,protein expression and differentiation ability of BMSCs in serum-free medium was similar to those of neural stem cells.The data support the hypothesis that adult bone marrow contains stem cells capable of differentiating into neural cells,the serum-free media make BMSCs overcome their mesenchymal commitment,showing the phenotypes for neural stem cells.

Objective To investigate the effect of dexmedetomidine (Dex)combined with ischemic preconditioning on liver ischemic reperfusion injury in rats in order to explore its possible mechanism. Methods Sixty healthy male SD rats weighing (251 ±18)g,were randomly (random number)divided into five groups (n =12 in each):sham-operation group (Group S:operation without ischemia), ischemia-reperfusion group (Group IR:hepatic pedicle occlusion for 30 min and reperfusion for 6 h), dexmedetomidine preconditioning group (Group Dex: dexmedetomidine 25 μg/kg was given intra-peritoneally at 30 min before operation),ischemic preconditioning group (Group IP:10 min ischemia,10 min reperfusion,followed by hepatic IR)and Dex combined with ischemic preconditioning group (Group Dex +IP:Dex 25μg/kg was given intra-peritoneally at 30 min befor operation,10 min ischemia and 10 min reperfusion was given followed by hepatic IR).The hepatic inflow of blood stream was occluded for 30 min by Pringle maneuver to establish hepatic ischemic reperfusion injury (HIRI)rat model.At the end of reperfusion for 6 h,the concentration of ALT,AST and LDH (lactate dehydrogenase)in serum were measured.The liver histological changes were examined after HE staining.The liver cell apoptosis were examined by TUNEL. The expression of heme oxygenase-1 were examined by Westeren blot and immunohistochemistry.H2 O2 and GSH (r-glutamylcysteinylglycine ) in liver tissue were detected by spectrophotometer.Differences among the groups were analyzed by one-way analysis of variance (ANOVA) and Student-Newman-Keul test by using SPSS version 17.0 software.Differences were considered significant at P <0.05.Results Serum concentrations of ALT,AST and LDH in group IR,Dex,IP and Dex +IP were significantly higher than those in group S (P =0.000).And those biomarkers in group Dex,IP and Dex +IP,were significantly lower than those in group IR (P =0.000).Furthermore,those biomarkers in group Dex +IP,were significantly lower than those in group Dex and IP (P =0.000).There were no significant difference in the serum concentrations of ALT and AST between group Dex and IP (P =0.550, 0.771),and the serum level of LDH in group Dex was significantly lower than that in group IP (P =0.000).The liver histopathological score and apoptosis index were the lowest in group S and the highest in group IR,and those in group Dex and group IP were significantly lower than those in group IR (P =0.000),and those in group Dex +IP were significantly lower than those in group Dex and group IP (P =0.000),and there were no significant difference between group Dex and IP (P =0.704,0.661 ).The expression score of HO-1 was the lowest in group S and the highest in group Dex +IP,and that in group Dex and group IP was significantly lower than that in group Dex +IP (P =0.000,0.002),and that in group IR was significantly lower than that in group Dex and group IP (P =0.000),and there was no significant difference between group Dex and IP (P =0.099).In respect of H2 O2 level and GSH level in liver tissue, compared with group S,the H2 O2 levels in groups IR,Dex,IP and Dex +IP were significantly higher (P=0.000,0.000,0.000,0.001)while the GSH levels were significantly lower (P =0.000).Compared with group IR,the H2 O2 level in group Dex,IP and Dex +IP was significantly lower than that in group IR while the GSH level was significantly higher (P =0.000 ).The H2 O2 level in group Dex +IP was significantly lower than that in group Dex and IP while the GSH level was significantly higher (P =0.000).There were no significant difference between group Dex and IP (P =0.480,0.667).Conclusions Both of dexmedetomidine and ischemic preconditioning can protect liver from ischemia reperfusion injury in rats to some extent,and the combined application of two gives better effects,which is attributed to the increasing expression of HO-1 to a certain extent.

OBJECTIVE: To construct and obtain ideal protein delivery vectors by researching the delivery efficiency and cytotoxicity to Hela cells using mPEG-PLGA-BSA-FITC-NPs. METHOD: The mPEG-PLGA nanoparticle was obtained through surface modification of PLGA with PEG, and deliver BSA-FITC into Hela cells in vitro. The positive cells were counted by Laser scanning confocal microscopy and the survival rate of Hela cells was calculated by MTT assay at different time points. RESULT: mPEG-PLGA-BSA-FITC-NPs shows the classic nanometer size, and the encapsulation efficiency reached 51. 2%. At the same time, the nanoparticles possess characteristics of slow release. By optimizing the delivery conditions, the highest efficiency of mPEG-PLGA-BSA-FITC-NPs was above 65.2%, and the cellular viability was about 85.7%. CONCLUSION mPEG-PLGA-BSA-FITC-NPs nanoparticles can successfully carry the target protein into cells as safe and effective as novel delivery materials of protein in vitro, and has shown slow release characteristics. The mPEG-PLGA-BSA-FITC-NPs provide ideal delivery vector for future application in clinical treatment of disease using nano-materials.
Drug Carriers ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; HeLa Cells ; Humans ; Nanoparticles ; Particle Size ; Polyesters ; Polyethylene Glycols ; Serum Albumin, Bovine

Objective: To explore the effect of mindfulness traits on sleep quality of college students, and to investigate the mediating mechanism of cognitive emotion regulation and anxiety for mindfulness trait factors, and sleep quality in college students. Methods: Totally 397 college students from five colleges were investigated using Five Facet Mindfulness Questionnaire, Cognitive Emotion Regulation Questionnaire, Self-Rating Anxiety Scale and Pittsburgh Sleep Quality Index.SPSS 22.0 version and Bootstrap method were used to analysis data and test intermediate effect. Results: (1)The total score of mindfulness trait and the score of describe, act with awareness and non-judge factors in mindfulness traits were(118.44±9.38), (24.82±3.87), (27.41±5.53) and (25.07±4.60), respectively.The total score of sleep quality was (5.57±2.62), and it was negatively related to the above three factors (r=-0.13--0.37, all P<0.05). (2)Anxiety played a complete mediating role between describe factor and sleep quality.Negative cognitive emotion regulation and anxiety played multiple mediating roles between the total score of mindfulness trait, the act with awareness factor and sleep quality.Negative cognitive emotion regulation and anxiety played a complete chain mediated role between non-judge factor and sleep quality. Conclusion The describe, act with awareness and non-judge factors in mindfulness traits can affect sleep quality through mediation of negative cognitive emotion regulation and / or anxiety.

Objective To observe the efficacy of microwire penetration combined with ozone intervention for fallopian tube obstructive infertility.Methods Data of 149 patients with fallopian tube obstructive infertility were retrospectively analyzed.Patients in group A(n=44)underwent conventional fallopian tube recanalization(FTR),in group B(n=51)underwent microwire penetration FTR,while those in group C(n=54)received microwire penetration FTR combined with ozone.The recalculation rate 1 month after treatment and natural pregnancy rate within 1 year after treatment were compared among groups.Results The recalculation rate 1 month after treatment of group A,B,C was 63.38%(45/71),80.22%(73/91)and 92.78%(90/97),respectively,increased in order among groups(all P＜0.05).The natural pregnancy rate within 1 year after treatment of group A,B,C was 20.45%(9/44),27.45%(14/51)and 48.15%(26/54),respectively,of group C was higher than of group A and B(both P＜0.05),not significantly different between group A and group B(P=0.427).Conclusion Microwire penetration combined with ozone intervention had better efficacy than conventional FTR and microwire penetration FTR alone for treating fallopian tube obstructive infertility.

Objective:To explore the ventilation strategy of using pressure controlled ventilation-volume guaranteed (PCV-VG) mode combined with personalized positive end expiratory pressure (PEEP) during surgery, and its impact on oxygenation function and incidence of postoperative pulmonary complications (PPCs) in patients undergoing brain tumor surgery.Methods:Sixty patients who underwent elective brain tumor surgery at the Linyi People′s Hospital from January 2023 to June 2023 were selected. The patients were randomly divided into PCV-VG ventilation mode group (T group) and volume controlled ventilation (VCV) mode group (C group) using a random number table method, with 30 patients in each group. One patient was excluded from group T due to changes in the condition, and 29 patients were actually included. Group T adopted PCV-VG ventilation mode and searched for the maximum dynamic lung compliance (Cdyn) by titrating PEEP. The PEEP corresponding to the maximum Cdyn value was the optimal PEEP, which was maintained until the end of mechanical ventilation. Group C adopted a ventilation mode of VCV plus 5 cmH 2O fixed value PEEP. All patients underwent arterial blood gas analysis before anesthesia induction (T 1), 15 minutes after setting ventilation mode (T 2), and 15 minutes after extubation (T 3), recording arterial oxygen partial pressure (PaO 2), carbon dioxide partial pressure (PaCO 2), lactate (Lac), and blood glucose (Glu), and calculating oxygenation index (PaO 2/FiO 2). The levels of white blood cells (WBC), C-reactive protein (CRP), and procalcitonin (PCT) were recorded before and 72 hours after surgery. The incidence of postoperative PPCs at 72 hours was observed. Results:There was a statistically significant difference in the grouping effect and time effect of PaO 2 and PaCO 2 between group C and group T (all P<0.05), while there was no statistically significant difference in the interaction effect (all P>0.05); The differences in grouping effects, time effects, and interaction effects of PaO 2/FiO 2 between group C and group T were statistically significant (all P<0.05); The average value of the optimized PEEP obtained by T-group titration was 7.48 cmH 2O, corresponding to an average platform pressure (Pplat) of 14.90 cmH 2O, and an average value of 53.37 ml/cmH 2O corresponding to the maximum Cdyn value. Compared with the Pplat and Cdyn corresponding to the fixed value of 5 cmH 2O PEEP in Group C, the Pplat corresponding to the optimized PEEP in Group T was lower than that in Group C ( P<0.05), and the Cdyn was higher than that in group C ( P<0.05). The CRP level and incidence of PPCs in group T after 72 hours of surgery were significantly lower than those in group C (all P<0.05). Conclusions:Compared with VCV, the ventilation mode of PCV-VG combined with personalized PEEP can improve intraoperative lung ventilation and oxygenation function in patients with brain tumors, reduce the occurrence of pulmonary inflammation 72 hours after surgery, and lower the incidence of PPCs.

Objective: To investigate the efficacy and safety of Rivaroxaban for elderly patients with thrombotic diseases. Methods: This was a retrospective study.A total of 301 elderly patients taking Rivaroxaban from October 2012 to November 2017 at the Second Medical Center of the Chinese PLA General Hospital were consecutively selected.The ages ranged from 60 to 102 years, with an average age of(86.5±8.4)years.Anticoagulation regimens were developed based on comprehensive evaluation of indications, creatinine clearance, ischemia and bleeding risk.Patients were divided into a Rivaroxaban 2.5-5.0 mg/d group(n=72), a 10.0 mg/d group(n=205), and a 15.0-20.0 mg/d group(n=24). Hepatic function, renal function, and coagulation indexes were measured before and after the administration of Rivaroxaban.Fatal bleeding, cardiovascular deaths, all-cause deaths, non-fatal bleeding and thromboembolic events were recorded during the follow-up period. Results: The average dose of Rivaroxaban was(9.3±3.0)mg/d, and the minimum dose was 2.5 mg/d.The average follow-up time was(14.9± 13.9)months and the longest follow-up time was 48 months.One patient had intracranial bleeding.Twenty patients(6.6%)died with a cumulative incidence of 25.2%, three(1.0%)died of cardiac events, and 55.0% died of pneumonia and multiple organ failure.Forty patients(13.3%)had non-fatal hemorrhagic events with a cumulative incidence of 42.4%.Seven patients(2.3%)had thromboembolic events with a cumulative incidence of 16.0%, including 2 cases of non-fatal myocardial infarction, 3 cases of cerebral infarction and 2 cases of deep vein thrombosis.After treatment, levels of prothrombin time and fibrinogen significantly increased while levels of D-dimer significantly deceased(P<0.05). Conclusions Compared with previous reports, low-dose Rivaroxaban is safe and effective for elderly patients with thrombotic diseases.However, the risk of bleeding and ischemia should be comprehensively evaluated, and appropriate doses of Rivaroxaban should be selected individually.