Objective To investigate the effects of postoperative sleep deprivation on the expression of choline acetyltransferase (ChAT) in hippocampi of aged rats.Methods Forty-eight male Wistar rats,aged 20 months,weighing 500-600 g,were randomly divided into 4 groups (n =12 each) using a random number table:control group (group C) ; operation group (group O) ; sleep deprivation group (group S) ; postoperative sleep deprivation group (group OS).Sleep deprivation was induced in the rats by housing them on small platforms over water.They fell into the water if they lost muscle tone.All the rats had free access to food and water.In group OS,splenectomy was performed,and all the rats underwent 24 h sleep deprivation after the rats were awake.All the rats underwent 24 h sleep deprivation at the corresponding time point in group S.Morris water maze test was carried out at 24 h after operation.The number of ChAT positive cells in the hippocampal CA1 region was counted after completion of Morris water maze test.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,and the frequency of crossing the original platform and the number of ChAT positive cells in the hippocampal CA1 region were decreased after operation in O and OS groups,and no significant changes were found in the parameters mentioned above in group S.Compared with group O,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,the frequency of crossing the original platform was decreased,and there was no significant difference in the numberof ChAT positive cells in the hippocampal CA1 region in group OS,and no significant changes were found in the parameters mentioned above in group S.Compared with group S,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,and the frequency of crossing the original platform and the number of ChAT positive cells in the hippocampal CA1 region were decreased after operation in group OS.Conclusion The mechanism by which postoperative sleep deprivation induces cognitive decline is not related to the expression of ChAT in hippocampi of aged rats.

Background Retinal neovascularization (RNV) is one of the major causes of blindness worldwide,but the pathogenic mechanism of this disease remains unclear, and therapeutic modalities need to be improved.Therefore,it is necessary to identify ocular protein markers with significant expression changes during RNV, thereby providing novel therapeutic targets for neovascular retinopathies.Objective This study was to investigate retinal vessel morphological characteristics and protein expression profiling in a mouse model of oxygen-induced retinopathy (OIR).Methods Forty four C57BL/6J mouse pups were randomly divided into normal control group and OIR group at postnatal day 7 (P7).The mice in the normal control group were raised under the normal air for 10 days.The mice of the OIR group were exposed to (75±2)％ oxygen for 5 days.The mother mice were alternated between the two groups every day.The mice of the OIR group returned to normal air at P12.Fluorescein isothiocyanate-dextran (FITC-dextran) was retrobulbarly injected at P17 mice from both groups, and the retinal flatmounts were prepared after fixation.The FITC-dextran-labeled retinal vessels were observed and quantified;the paraffin sections of eyeballs were processed for hematoxylin and eosin staining,and the number of pre-retinal vascular cell nuclei was quantified.The total proteins were extracted from the eyes, and the expression profiling was analyzed by a customized protein array and verified by Western blot and ELISA.Results The retinal flatmounts labeled with FITC-dextran showed that the peripheral retinal microvessels in the OIR group were tortuous, disorganized with neovascular buds,and the vascular obliteration was prominent in the center of retina.Contrastly,the vessels were smooth,organized, and evenly distributed in the normal control group.The percentage of vascular obliteration area in the OIR group was (25.53±2.16)％ ,which was significantly higher than (0.66±0.36)％ in the normal control group (t=-27.61 ,P＜ 0.01).The number of pre-retinal vascular cell nuclei,as revealed by hematoxylin and eosin staining, was (28.41 ± 3.97)/slide in the OIR group, which was substantially higher than (0.16±0.31)/slide in the normal control group (t =-54.42,P＜0.001).Protein array showed that 10 out of the 62 examined pro-inflammatory, pro-angiogenic cytokines exhibited more than 1.5-fold expression changes,including 3 up-regulated cytokines and 7 down-regulated cytokines;4 cytokines showed more than 2-fold expression changes,in which 3 cytokines were down-regulated and 1 cytokine was up-regulated.The differential expressions were verified by Western blot and ELISA.The expression trends of platelet factor 4 (PF-4), vascular endothelial growth factor A (VEGF-A), selectin P (SELP), vascular cell adhesion molecule 1 (VCAM-1) ,soluble tumor necrosis factor receptor Ⅱ (sTNF-RⅡ) and chemokine C-X-C motif ligand 16 (CXCL16) were consistent with those revealed by protein array.PF-4,VEGF-A and SELP were up-regulated,and the other 3 were significantly down-regulated (all at P＜0.05).Conclusions Differential expression patterns of the cytokines,including PF-4,VEGF-A, SELP, VCAM-1, sTNF-RⅡ and CXCL16, are identified between normal and OIR mouse eyes.These differential expression patterns suggest that under the condition of OIR,the platelet system is activated,and proinflammatory factors are down-regulated.PF-4 might become a new target for VEGF-independent therapeutic strategy against RNV.

BACKGROUND:Previous studies have documented that,the increase of anti-cardiolipin(aCL) antibody titer has an obvious positive relaltionship with the vascular thrombosis,thrombocytopenia and repeated abortion in the patients with systemic lupus erythematosus and antiphospholipid syndrome,but there is little information on the aCL antibodies in lupus nephritis(LN).OBJECTIVE:To ascertain the preyalence and significance of aCL antibodies in Chinese patients with LN.DESIGN:Prospective follow-up study of one sample.SETTING:Department of Rheumatology in Xiangmihu Branch of Shenzhen Fourth People's Hospital,Shenzhen Institute of Rheumatology in Guangdong Medical College.PARTlCIPANTS:The study was performed in 97 LN Patients consecutively recruited in the Department of Rheumatology in Xiangmihu Branch of Shenzhen Fourth People's Hospital between March 2001 and October 2003.All the included patients met the revised criteria of American College of Rheumatology for the diagnosis and classification of LN.And they all knew the fact saying yes.METHODS:The clinical data and auxiliary examination result were recorded when hospitalizalion.The aCL antibodies were measured by the enzyme-linked immunosorbent assay,and Were considered as positive if over 100 U/mL.High-dose oral administration of prednisonc combined with cyclophosphamide intravenous pulse therapy were applied for inducing release.The curative effect was remained by using azathioprine and prednisone at a decreasing dose.Meanwhile the complications such as hypertension,hyperlipemia and arthralgia were prevented by drugs.All the patients had routine visits at six-month intervals for a total of 3 years,Clinical and seroIogic manifestations of Patients with LN were tested and recorded regularly.MAIN OUTCOME MEASURES:Gender,age,systemic lupus erythematosus disease activity index,clinical manifestations,vascular thrombosis,pregnancy outcome and renal function.RESULTS:All the 97 LN Patients were included in the study.and 83 of them entered the result analysis while the other 14 cases were lost.The overall prevalence of aCL antibodies in 97 subjects was 39%(38 cases).Hypertension,thrombocytopenia and Raynaud's phenomenon were more frequent in LN Patients with aCL antibodies.The aCL IgG antibody-positive Patients showed a greater risk for the occurrence of vascular thrombosis;Pregnancy morbidity of miscarriages,premature birth,fetal death and the probabmty of developing irreversible renal function deterioration occurred at a greater frequency in aCL antibody-positive patients.CoNCLUSIoN:The prevalence of aCL antibodies in LN Patients is 39%.A higher incidence of hypertension.thrombocytopenia and Raynaud's phenomenon is found in patients with aCL antibodies.Detection of aCL antibodies in Patients with LN may be usefol to predict the development of vascular thrombosis,pregnancy morbidity and irreversible chronic renal function deterioration.

ObjectiveTo investigate the treatment of Mirizzi syndrome (MS) by ERCP、laparoscopy and choledochoscopy. MethodsIn this study 12 cases were confirmed intraoperatively as with MS from July 2005 to June 2009. Patients were treated by ERCP,laparoscopy and choledochoscopy according to the Csendes Classification. ResultThere were 7 MS patients complicating common bile duct stones among all 12 MS cases.There were 8 cases of Type Ⅰ Csendes MS,3 cases of Type Ⅱ and 1case of Type Ⅲ.11cases were treated by the ERCP、laparoscopy and choledochoscope.4 cases was treated by primary closure of common bile duct in laparoscopy,all the patients were cured.The case of type Ⅲ with T tube placed for stone caused bile duct injury had no stricture of the common bile duct as demonstrated by postoperative follow-up cholangiography.ConclusionsERCP,laparoscopy and choledochoscopy are effective for the treatment of Mirizzi syndrome.

Objective To investigate the inhibitory effect of alantolactone on glioma C6 cells and its possible mechanism. Methods Rat glioma cell line C6 cells were cultured in vitro. The effect of AL on the viability of C6 cells was detected by MTT assay. The effect of AL on migration and invasion of C6 cells was assessed by scratch test and Transwell chamber assay. Flow cytometry was used to detect the effect of AL on the induction of apoptosis in C6 cells, and the effects of AL on mitochondrial membrane potential in C6 cells was detected using JC-1 fluorescent probe. Western blot was used to detect the expression of related proteins in C6 cells after AL treatment. Results AL significantly inhibited the proliferation of C6 cells in a time-and dose-dependent manner. After AL treatment for 24 hours, the migration and invasion of C6 cells were significantly inhibited, N-cadherin protein was significantly decreased and E-cadherin protein was significantly increased, the number of apoptotic cells increased obviously while the mitochondrial membrane potential decreased significantly. The protein expressions of PARP/ cleaved caspase-3/cleaved caspase-9 were significantly up-regulated. Conclusions AL inhibits the proliferation of glioma C6 cells by inhibiting their migration and invasion through regulating the protein expression of cadherin and inducing apoptosis in C6 cells by regulating the cytochrome C/caspase signaling pathway.

OBJECTIVETo explore clinical effects of suturing-assisted locking plate in treating elderly proximal humeral fractures.

METHODSFrom January 2005 to January 2013, 55 elderly patients with three- and four-part fractures of proximal humeral fractures were divided into treatment group and control group. In treatment group, there were 31 patients including 12 males, and 19 females aged from 65 to 85 with an average of (74.00±5.42) years old, and treated with suturing-assisted locking plates; 19 patients were Neer 3-part fractures, and 12 patients were Neer 4-part fractures of proximal humerus; 23 patients were suffered from low-energy injuries and 8 patients were caused by high-energy injuries. In control group, there were 24 patients including 7 males, and 17 females aged from 65 to 85 with an average of (72.79±5.34) years old, and treated with locking plates; 16 patients were Neer 3-part fractures, and 8 patients were Neer 4-part fractures of proximal humerus; 17 patients were suffered from low-energy injuries and 7 patients were caused by high-energy injuries. Operative time, blood loss during operation, and bone healing time between two groups were observed and compared. Postoperative Neer scoring were used to evaluate recovery of shoulder joint function.

RESULTSAll patients were followed up from 6 to 24 months with an average of 16.1 months. In treatment group, blood loss was (495.806±143.150) ml, function of Neer scoring was 22.645±2.443, range of action was 18.194±2.613, anatomy was 7.935±1.504 and total score of Neer scoring was 77.161±8.335; while in control group, blood loss was (641.667±169.851) ml, function of Neer scoring was 13.958±1.989, range of action was 13.083±2.165, anatomy was 5.500±1.978 and total score of Neer scoring was 58.792±7.313. There were sigificant difference between two groups in these indexes.

CONCLUSIONSuturing-assisted locking plate for the treatment of proximal humerus fractures in elderly, has advantages of less blood loss, simple fracture reduction and rapid recovery of shoulder joint, and is a effective method.

Aged ; Aged, 80 and over ; Bone Plates ; Case-Control Studies ; Female ; Humans ; Male ; Recovery of Function ; Shoulder Fractures ; physiopathology ; surgery ; Shoulder Joint ; physiopathology ; Sutures

Objective To observe the effect of lentiviral vector-mediated basic fibroblast growth factor (bFGF) gene transfection on the biological characteristics of rabbit bone marrow stromal cells(BMSCs)under in vitro culture conditions. Methods BMSCs were obtained by density gradient centrifugation and adherence screening. The bFGF gene was transfected into BMSCs by lentiviral vector and divided into bFGF transfection group,empty virus group and untransfected group according to the transfection conditions.After transfection,the morphology,expressions of bFGF mRNA and protein, cell proliferation,cell cycle and alkaline phosphatase(ALP)activity were observed in three groups of cells. Results High density BMSCs were successfully obtained by density gradient centrifugation and adherence screening.After transfection of BMSCs with bFGF gene, the cell morphology showed no significant changes, while the expressions of bFGF mRNA and protein were significantly increased, the cell proliferation curve shifted upward, the proportion of proliferating cells increased,and the activity of ALP was significantly enhanced.There were significant differences between three groups(P<0.05).Conclusion The rabbit bFGF gene is successfully introduced into the BMSCs cultured in vitro by lentiviral vector, and the target gene is stably expressed.The expression of bFGF can promote the proliferation and osteogenic differentiation of BMSCs.

BACKGROUNDThe treatment for long bone defects has been a hot topic in the field of regenerative medicine. This study aimed to evaluate the therapeutic effects of calcium sulfate (CS) combined with platelet-rich plasma (PRP) on long bone defect restoration.

METHODSA radial bone defect model was constructed through an osteotomy using New Zealand rabbits. The rabbits were randomly divided into four groups (n = 10 in each group): a CS combined with PRP (CS-PRP) group, a CS group, a PRP group, and a positive (recombinant human bone morphogenetic protein-2) control group. PRP was prepared from autologous blood using a two-step centrifugation process. CS-PRP was obtained by mixing hemihydrate CS with PRP. Radiographs and histologic micrographs were generated. The percentage of bone regenerated bone area in each rabbit was calculated at 10 weeks. One-way analysis of variance was performed in this study.

RESULTSThe radiographs and histologic micrographs showed bone restoration in the CS-PRP and positive control groups, while nonunion was observed in the CS and PRP groups. The percentages of bone regenerated bone area in the CS-PRP (84.60 ± 2.87%) and positive control (52.21 ± 4.53%) groups were significantly greater than those in the CS group (12.34 ± 2.17%) and PRP group (16.52 ± 4.22%) (P < 0.001). In addition, the bone strength of CS-PRP group (43.10 ± 4.10%) was significantly greater than that of the CS group (20.10 ± 3.70%) or PRP group (25.10 ± 2.10%) (P < 0.001).

CONCLUSIONCS-PRP functions as an effective treatment for long bone defects through stimulating bone regeneration and enhancing new bone strength.

Animals ; Bone Regeneration ; drug effects ; Calcium Sulfate ; pharmacology ; Male ; Platelet-Rich Plasma ; Rabbits

OBJECTIVETo describe the rule of the schwann cell proliferation after peripheral nerve injury in detail and to discover the effect of neuroanastomosis.

METHODSThe model of rat sciatic injury was made, with neuroanastomosis on the left side and right side untreated. Then draw the materials 24, 48 h, 4, 7, 14, 21 d after surgery. And immunohistological stain counted the schwann cell per view with Qwin software of Leica Ltd.

RESULTSThe number of schwann cell increased obviously 4 d after surgery and reached the peak in 7 d. Then it fell down and the neuroanastomosis group changed slower and fibroblast hyperplasia in the untreated.

CONCLUSIONSThe axon support is essential for the schwann cell. The precise rule is help for study on neurotrophic factor.

Anastomosis, Surgical ; Animals ; Cell Proliferation ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Schwann Cells ; cytology ; Sciatic Nerve ; injuries ; pathology ; surgery ; Time Factors

OBJECTIVETo rudiment analyze the different intervention factors affecting the spinal cord injury functional recovery initially through prospective study project.

METHODSDesigned prospective study project without clinical treatment intervention and admitted 62 spinal cord injury cases into observation group from Dec 2006 to Dec 2007. The internalized standard included acute spinal cord injury within 1 week, aged from 18 to 65 years old without sexuality difference and diagnosed as type A injury: total spinal cord injury or type B injury: non-total spinal cord injury (without movement function below the injury plane) according to clinical physical examination and combined MRI or CT examination. All the 62 cases were followed up. The spinal cord function were estimated at the time of arriving hospital, 1, 3, 6 months after injury during the following up time according to the American Spine Injury Association standard (Revised at 2000) and functional independence measure (FIM) score. The intervention factors affecting the spinal cord injury functional recovery were analyzed.

RESULTSAmong the 62 cases, male 60 cases and female 2 cases, aged from 18 to 41 years old, mean age 24 years old, 29 cases can be classified into type A injury and 33 cases can be classified into type B injury. There was no significant difference (Index including sense ASIA, motor ASIA and FIM) between operation group and non-operation group, namely conservatively composite treatment at all observation point in all type A injury cases (P > 0.05). There was significant difference (Index including sense ASIA, motor ASIA and FIM) between operation group and non-operation group at all observation point in all type B injury cases (P < 0.05); And there was significant difference (Index including sense ASIA, motor ASIA and FIM) between operated in 8 hours (< or = 8 h) group and beyond 8 hours (> 8 h) group at all observation point in all type B operated cases (P < 0.05).

CONCLUSIONSOperation and operation time were of no significant value for type A injury (total spinal cord injury), but considering the nursing convenience and the need for spinal stabilities, operation decompression and internal fixation can be chosen; Operation decompression should be performed as soon as quickly for type B injury (non-total spinal cord injury) in order to get better functional recovery.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Injury Severity Score ; Male ; Middle Aged ; Prospective Studies ; Recovery of Function ; Spinal Cord Injuries ; physiopathology ; surgery ; Treatment Outcome ; Young Adult