Objective To establish the model of amyotrophic lateral sclerosis (ALS) with selective motor neuron disorder by organotypic spinal cord cultures, and analyze the role of astrocyte in the pathagenisis of ALS.Methods Organotypic spinal cord cultures were prepared using lumbar spinal cord slices from 8-day-old SD rat pups. The threohydroxyaspartate (THA) was applied into culture medium to establish ALS organotypic spinal cord cultures model. Motor neurons survival was evaluated by monoclonal SMI-32 immunohistochemical staining and glial fibrillary acidic protein staining to show astrocyte survival.Results Compared with the control group, there was significantly astrogliosis in the anterior horn and surrounding white matter in THA 100 μmol/L group, and the level of gliosis was increased followed the elongation of THA interference time. With the increasing of the number of astrocyte, the morphology of astrocyte was changed.Conclusion There is significantly astrogliosis in the anterior horn and the time of astrogliosis is markedly earlier than the time of motor neuron loss in the ALS model intervented with THA.

BACKGROUND: Mesenchymal stem cells (MSCs) can differentiate into myocardial cells in vitro and in vivo, but the amount is small and directed differentiation rate is low.OBJECTIVE: To explore the effect of stem cell factor (SCF) on promotion of MSCs differentiating into myocardial cells.DESIGN: Opening experiment.SETTING: Institute of Cardiovascular Disease, Xianning College and Department of Cardiology, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was performed at the Experimental Center of Institute of Cardiovascular Disease, Xianning College from October 2003to August 2004. A total of 20 infant SD rats aged 1-2 days were selected for culture of myocardial cells. Another 25 clean adult SD rats were selected and randomly divided into SCF group, blank control group with 12 rats in each group. The left one rat was used for MSCs extraction, culture and purification.METHODS: ①MSCs were labeled with 4', 6-diamidino-2-phenylindole,dihydrochloride (DAPI) before co-culture. SCF was given into rats of the SCF group by subcutaneous injection successively for 5 days, 20 μg/kg per day. Isolated MSCs were co-cultured with myocardial cells that had been cultured for 3 days. Saline of the same volume was given in the blank control group by subcutaneous injection. MSCs without any intervention were co-cultured with myocardial cells that had been cultured for 3 days.②Expressions of MHC α/β and troponin T were recorded and measured with digital micro-camera shot and immunofluorescence technique, respectively. Percentage of DAPI labeled MSCs differentiating into myocardial cells was measured.MAIN OUTCOME MEASURES: ①Growth of MSCs, ②detection of labeling rate of DAPI on MSCs, ③analysis of activity and purity of co-cultured myocardial cells, and ④differentiation of MSCs into myocardial cells after co-culture.RESULTS: ①Bone marrow cell suspension was inoculated in plastic petri dish. Round cells scattered at the bottom of bottle. At hour 24 a few long fusiform shape adhered cells appeared following liquor change, and at day 4 many fusiform shape adhered cells appeared, which entered logarithm increased period. At days 8-12 80% were confluence. The proliferation of passage cells was more rapid. With liquor change and passage, the MSCs were purified gradually. ②Labeling rate of DAPI on MSCs was 100%. ③At 3-day in vitro culture of myocardial cells the percentage of beat cells was 63%, with beat frequency of 40-60 times per minute. Percentage of positive cells of troponin T expression was 75% examined with immunocytochemical technique. ④At co-cultured days 2 and 3, positive percentage of DAPI labeled MSCs expressing MHC α/β was significantly higher in the SCF group than in the blank control group (P ＜ 0.01). At the co-cultured days 3, 4 and 5, positive percentage of DAPI labeled MSCs expressing troponin T was obviously higher in the SCF group than in the blank control group (P ＜ 0.01).CONCLUSION: SCF has markedly accelerating effect on proliferation and differentiation of MSCs.

Objective To evaluate operative management of complex hypospadias.Methods Twenty-one cases with complex hypospadias were reviewed.Thiresch procedure at 12 cases were taken.Island scrotal septal flap urethroplasty at 7 cases.Snodgrass procedure at 1 case.Mathieu procedure at 1 case.Results Fifteen cases were satisfied without fistula and stricture.Four cases with fistula,1 case with chordee,urethral meatus stricture in 1 case.Overall,the complication rate was 28.5%.Conclusions Selection of surgical procedure should according to the different case circumstance.It′s essential that reduce separative procedure,protect blood supply and carefully manipulation to improve successful rate.

AIM:To investigate the effect of pretreatment of stem cell factor(SCF) and granulocyte colony-stimulating factor(G-CSF) on the proliferation and the differentiation of mesenchymal stem cells(MSCs) into cardiomyogenic cells.METHODS:The MSCs,isolated primarily from bone marrow,and purified by passage culture,were obtained from the adult rats of four groups:the rats were pretreated by 5 daily injections of SCF;the rats were pretreated with G-CSF;the rats were pretreated with SCF and G-CSF;the rats were treated without any intervention.The 4th passage of MSCs was labeled by DAPI and cellular cycle analysis was conducted by flow cytometry before co-culture.The neonatal rat cardiomyocytes cultured for 3 days were co-cultured with DAPI-MSCs.The percentage of the differentiation of MSCs into cardiomyogenic cells during the five co-culture days was analyzed.The morphologic changes of MSCs and the proteins expression of cardiac myosin heavy chain(MHC) and troponin T(TnT) were recorded respectively with digital microscope camera system and immunofluorescence technique.The percentage of the differentiation of MSCs into cardiomyogenic cells was also calculated.RESULTS:The percentage of MSCs in G0/G1 phase in SCF/G-CSF group was significantly lower than that in SCF group,G-CSF group and the control group.The percentage of MHC protein-positive MSCs in SCF/G-CSF group was markedly higher than that in SCF group,G-CSF group and the control group,and that in SCF group and G-CSF group was significantly higher than control group.The percentage of TnT protein-positive MSCs in SCF/G-CSF group,SCF group and G-CSF group was significantly higher than that in control group.CONCLUSION:SCF and G-CSF show the ability to stimulate the proliferation of MSCs and induce MSCs to differentiate into cardiomyocytes.The combination of using SCF and G-CSF is more effective than using only SCF or G-CSF.

Spermiogenesis is a complex process of differentiation and morphologic alteration, in which sperm acrosome formation is an important stage. Acrosome is an essential component of the sperm head, which develops in four distinct phases: Golgi, cap, acro- somal, and maturation, each supported by precise and orderly regulation of various genes. The regulatory genes which act on Golgi ap- paratus include GOPC, Hrb, SPATA16, PICK1, and CK2α', those involved in the cap phase are Fads2, syntaxin 2, Kdm3a, and UBR7, and participating in acrosomal and maturation phases are KIFC1, Rnf19a, and DPY19L2. The abnormalities of these genes may affect male fertility by influencing the connection of the nuclear dense lamina and acroplaxome with the nuclear membrane and then the fusion and transportation of vesicles. This review focuses on the genes involved in different phases of acrosome formation.
Acrosome ; physiology ; Animals ; Golgi Apparatus ; Male ; Mice ; Sperm Head ; physiology ; Spermatids ; growth & development ; Spermatogenesis ; genetics ; Spermatozoa ; growth & development

Objective To investigate the effects of penehyclidine hydrochloride combined with ulinastatin on postoperative cognitive function in the patients undergoing thoracic surgery.Methods One hundred and twenty patients undergoing lung cancer thoracotomic radical resection were randomly divided into hydrochloride penehyclidine composite ulinastatin group(HU group),hydrochloride penehyclidine group(H group),ulinastatin group(U group)and control group(C group).The arterial blood was collected for detecting OI,TNF-α,IL-6 and IL-8.The serum levels of S-100β and NSE were detected.The MMSE scores were evaluated.Results Compared with the H group and U group,the levels of TNF-α at T3-4 in the HU group were decreased,and the levels of IL-6 and IL-8 at T2-4 were decreased,while OI was increased(P＜0.05).Compared with the H group and U group,the serum levels of S-100β and NSE at T5-8 in the HU group were decreased(P＜0.05);compared with the H group and U group,the MMSE scores at T6-7 in the HU group were increased(P＜0.05).Conclusion Penehyclidine hydrochloride combined with ulinastatin could reduce the inflammation reaction during one lung ventilation in thoractomy and improves the postoperative cognitive function.

OBJECTIVETo establish sleeve gastrectomy(SG) rats model of obese type 2 diabetes mellitus(T2DM) for the research of hypoglycemic mechanism.

METHODSNine male Sprague-Dawley (6-week-old) rats were fed with high-sucrose and high-fat diet for 4 weeks, developing diet-induced obesity (DIO) rats model. The rats were then randomly divided into two groups. Six rats of them underwent sleeve gastrectomy(SG) as the sleeve gastrectomy group[SGG, body weight (471.8±17.9) g] and the other three rats underwent a laparotomy and stomach manipulation as the sham operative group[SOG, body weight (467.0±8.4) g]. The body weight, caloric intake and peripheral blood concentration of total ghrelin of rats were recorded after operation.

RESULTSThe weight of all the rats declined progressively after operation. The weight of the rats in SOG began to rise on the 5th postoperative day(POD) and regain their preoperative levels on the average 22nd POD. However, the weight of the rats in SGG began to rise slowly from the 9th POD, but was still lower than that of SOG[(487.4±10.1) g] and preoperative levels[(471.8±17.9) g] on the 28th POD[(420.1±18.6) g](P=0.001). Average caloric intake of rats in SGG was significantly lower than that of SOG after operation, but there was no significant difference between the two groups(P=0.121). The ghrelin level of SGG showed a continuous decreasing trend after intervention, decreased by 17.4% compared with the preoperative level (1595.1±14.4 ng/L) on the 28th POD[(1316.8±14.8) ng/L]. The ghrelin level of SOG did not change obviously before and after operation and both groups differ statistically(P=0.000).

CONCLUSIONSA SG rat model is successfully established. This model can be used for the further study of mechanism analysis of T2DM resolution after surgery.

Animals ; Diabetes Mellitus, Type 2 ; pathology ; Disease Models, Animal ; Gastrectomy ; methods ; Ghrelin ; blood ; Male ; Obesity ; blood ; surgery ; Rats ; Rats, Sprague-Dawley

Objective To compare the efficacy of different target concentrations of etomidate in combination with midazolam,fentanyl and rocuronium used to induce anesthesia for tracheal intubation.Methods Eighty ASA Ⅰ or Ⅱ and Mallampati Ⅰ or Ⅱ patients of both sexes,aged 25-50 yr,weighing 57-76 kg,scheduled for elective non-cardiac surgery under general anesthesia,were randomly allocated into 4 groups according to the target effect-site concentration of etomidate (n =20 each) ∶ 0.5 μg/ml group (group E0.5),0.7 μg/ml group (group E0.7),0.9μg/ml group (group E0.9) and 1.1 μg/ml group (group E1.1).The patients were unpremedicated.Anesthesia was induced with midazolam 0.05 mg/kg,fentanyl 3 μg/kg,rocuronium 0.6 mg/kg and etomidate given by target-controlled infusion.When the effect-site concentration of etomidate reached 0.5,0.7,0.9 or 1.1 μg/ml,endotracheal intubation was performed.Auditory evoked potential index was recorded before induction of anesthesia (baseline),immediately before intubation,during insertion of the laryngoscope,and at 1,3 and 5 min after intubation.Myoclonus,injection pain,the requirement for vasoactive agents and burst suppression (BS) were recorded during induction of anesthesia.Results Compared with group E0.5,the requirement for urapidil was significantly decreased in group E0.7,the requirement for esmolol and urapidil was significantly decreased and the incidence of BS was increased in group E0.9,the requirement for esmolol and urapidil was significantly decreased,and the requirement for atropine and ephedrine and incidence of BS were increased in group E1.1 (P ＜ 0.05).The incidence of BS was significantly higher in group E0.9,and the requirement for atropine and incidence of BS were significantly higher in group E1.1 than in group E0.7 (P ＜ 0.05).The incidence of BS was significantly higher in group E1.1 than in group E0.9 (P ＜ 0.05).There was no significant difference in auditory evoked potential index and incidences of myoclonus and injection pain among the four groups (P ＞ 0.05).Conclusion The optimum target concentration of etomidate is 0.7μg/ml when combined with midazolam,fentanyl and rocuronium used to induce anesthesia.

Forty five patients with acute cerebral infarction were randomized to two groups: in treatment group patients received local subhypothermia and conventional therapy, in control group patients received conventional therapy only. Clinical outcome was assessed by the National Institutes of Health Stroke Scale (NIHSS) on admission and at 7, 14 and 30 d after treatment. Serum neuron specific enolase (NSE), nitrogen monoxide ( NO ) , superoxide dismutase (SOD), interleukin-6 (IL-6 ) and intercellular adhesion molecule-1 (ICAM-1) were detected on admission and at 7,14 d after treatment The study showed that NIHSS scores of treatment group on 14, 30 d were lower than those of control group ( P ＜ 0. 05 ). Serum NSE, NO, IL-6 and ICAM-1 levels significantly decrease; while serum SOD levels increased (P ＜ 0. 05). In conclusion, local subhypothermia therapy can inhibit inflammatory reaction, reduce oxygen free radical formation and improve neurological function in patients with acute cerebral infarction.

Objective To investigate the effects of monocytes on phenotypic changes of human proximal tubular HK-2 cells and the mechanism.Methods Monocytes were co-cultured with HK-2 cells.Morphological changes of HK-2 cells were detected by inverted phase contrast microscope.Expressions of E-cadherin,α-SMA and fibronectin were assessed by RT-PCR,Western blotting and immunocytochemical staining.Flow cytometry techniques was applied to evaluate intercellular cell adhesion molecule-1 (ICAM-1) expression on HK-2 cells.The intracellular signal was investigated by gene microarr ay.Results The typical epithelial cell morphology of HK-2 cells disappeared after co-culture with monocytes,accompanied by decreased E-cadherin expression and increased α-SMA and fibronectin expression (all P＜0.05).The expression of ICAM-1 on HK-2 cells was increased by monocytes stimulation.Interestingly,administration of CD18 antibody directly inhibited the phenotypic change of HK-2 cells.Furthermore,NF-κB signaling might be critical in mediating this process,and blockade of this signaling pathway could inhibit 1CAM-1 expression and epithelial mesenchymal transition (EMT) formation.Conclusion Monocytes can directly induce EMT of HK-2 cells via up-regulating ICAM-1 through NF-κB signaling pathway.