OBJECTIVETo search for an optimal protocol and freezing conditions for the cryopreservation of microamount round spermatids of the mouse.

METHODSWe compared the survival rates of frozen-thawed microamount round spermatids of the mouse achieved by vitrification or standard slow freezing with different concentrations of glycerol (5, 7, or 9%) and different lengths of equilibrium time (0, 15, 30, 45, or 60 min).

RESULTSUnder the conditions of 7% glycerol and 30 min equilibrium, both vitrification and standard slow freezing achieved high survival rates of spermatids, and the former obtained an even higher rate than the latter ([72.9 ± 15.4]% vs [58.2 ± 17.7]%, P < 0.05).

CONCLUSIONA high rate of frozen-thawed microamount round spermatids of the mouse can be achieved by vitrification under the conditions of 7% glycerol and 30 min equilibrium.

Animals ; Cell Survival ; Cryopreservation ; methods ; Cryoprotective Agents ; administration & dosage ; pharmacology ; Glycerol ; administration & dosage ; pharmacology ; Male ; Mice ; Spermatids ; Time Factors ; Vitrification ; drug effects

Leiomyomas are benign soft tissue swellings of smooth muscle origin, most commonly found in the uterus. Extra uterine leiomyomas presenting as an abdominal mass is often a diagnostic challenge as such occurrence is rare. We present a rare case of primary abdominal wall leiomyoma, and highlight the importance of laparoscopic approach in the diagnosis and treatment of such tumour.
Leiomyoma

OBJECTIVETo study the promoting effects of the epidermal growth factor (EGF) and stem cell factor (SCF) on the proliferation and differentiation of spermatogenic cells in mice.

METHODSWe cocultured in vitro spermatogenic cells of male mice aged 7 - 8 days in the medium with EGF and/or SCF at the concentrations of 5, 10, 20, 40 and 100 ng/ml, respectively. Then we observed the survival rate and morphological changes of the spermatogenic cells, detected the expressions of the pachytene-specific phosphoprotein gene (P19) and haploid sperm cell-specific transition protein gene (TP1), and analyzed the ploidy of the cells.

RESULTSAfter cocultured with EGF or SCF for 2 - 4 days, the spermatogenic cells began to proliferate in masses or chains in all concentration groups, most obviously in the 20 ng/ml EGF and 40 ng/ml SCF groups. At 7 days, both the number and survival rate of spermatogenic cells were significantly higher in the 20 ng/ml EGF and 40 ng/ml SCF groups than in the others (P < 0.05), and meanwhile, the P19/TP1 ratio was obviously decreased and the rate of haploid sperm markedly increased in the 40 ng/ml SCF group (P < 0.05). Combination of EGF and SCF remarkably promoted the proliferation of the spermatogenic cells (P < 0.05).

CONCLUSIONBoth EGF and SCF could increase the number and survival rate of spermatogenic cells. SCF could promote the formation of haploid sperm, and the combination of the two cytokines could enhance the effect on the proliferation of spermatogenic cells.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chromosomal Proteins, Non-Histone ; metabolism ; Epidermal Growth Factor ; pharmacology ; Male ; Mice ; Spermatocytes ; cytology ; drug effects ; Stem Cell Factor ; pharmacology

Background Recently,there were many studies on corneal innervations during mammalian development.However,there were fewer studies on discussing corneal innervations before and after mouse eye openings.Objective The present study was to investigate the change in the regulation of corneal nerve fiber length and density before and after mouse eye openings to offer a basis for clinical research in human.Methods Thirty SPF C57BL/6 mice were divided into postnatal 1 day(P1 d),P7 d,P13 d(1 day before eye opening),P14 d(eye halfopened),P17 d(1 day after eye opening)and P23 d(7 day after eye opening)groups,with 5 mice and 10 eyes for each group.Entire corneal stretches were prepared and immunostaining with an anti-neuron-specific β-Ⅲ tubulin antibody was performed to label the corneal nerve fibers.Confocal microscopic pictures from the corneal dorsal-nasal region (DN),dorsal-temporal(DT),ventral-nasal region(VN)and ventral-temporal(VT)were taken using Delta Vision Core.From these pictures,the mouse corneal area,total length and density of nerve fibers in the 4 regions were calculated.The use of the animals complied with Statement of ARVO.Results Corneal areas of P1 d,P7d,P13 d,P14 d,P17 d and P23 d mice were(0.404±0.007),(1.362±0.154),(1.573±0.080),(1.603±0.046),(1.847±0.052),(2.445±0.798)mm2,respectively ; the total lengths of nerve fibers were(3.718±1.044),(19.065±3.350),(23.687±0.907),(27.309±2.477),(31.989±3.976),(41.214±1.573)mm,respectively ; the densities of nerve fibers were(9.592±1.138),(14.506±1.908),(15.088±1.241),(16.772±1.897),(16.821±2.102),(17.660±1.216)mm/mm2,respectively,all showing significant increases with age(F =22.906,P =0.000 ; F =0.424,P =0.000 ; F =2.375,P=0.000).A positive correlation of the increasing corneal areas and increasing lengths of nerve fibers was found(r=0.983,P＜0.01).Nerve fiber densities in the four corneal regions significantly increased with age(DN region:F =0.159,P =0.000 ; DT region:F =2.1 72,P =0.001 ; VN region:F =1.998,P =0.000 ; VT region:F=2.352,P=0.000).From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference(t =0.589,P =0.572); and the corneal nerve fiber densities in the DT region,VN region and VT region decreased by 4.6％,5.5％ and 0.1％,respectively,without significant difference from P14 d to P17 d(t=0.549,P=0.596;t=0.701,P=0.501 ;t=-0.100,P=0.919).Conclusions The development of nerve fibers in the whole cornea or the four corneal regions is influenced by eye opening in mouse to various extents.From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference.From P14 d to P17 d,the corneal nerve fiber densities in the DT region,VN region and VT region decrease by 4.6％,5.5％ and 0.1％,respectively,without significant difference.Afterwards,the growth of nerve fibers increased in pace and the growth rate is recovered.

The three immediate-early proteins of HSV-1, ICPO, ICP22, and ICP27, have specific and pivotal functions in transcriptional activation and inhibition, multiple regulatory and control processes of viral genes. In this paper, the expression and localization of these three proteins were studied in neuroblastoma cells using biochemical assays, and their possible and potential interactive functions are discussed. The data show that the three proteins are localized in different structures, specifically in the PML-NB-associated structure, which is a specific nuclear structure composed of many protein molecules and bound tightly to the nuclear matrix in neuroblastoma cells. The results suggest that the activating and suppressive functions of ICPs are mostly dependent on their transcriptional and regulatory roles, including the PML-NB-associated structure.

Objective To study the prognostic value of circumferential resection margin(CRM)status. Methods Specimens of 62 patients with rectal cancer,who underwent total mesorectal excision from December,2001 to June,2002,were examined using whole-mount sections.All patients were followed up for 4～60 months. Results Circumferential resection margin involvement(CRMI)was identified in 12 specimens.while only 4 of them were cancer positive in the resection margin.Analyzed by logistic regression test,the occurrence of CRMI was related to tumor differentiation [ P=0.015,Exp(B)=0.131]and lymph node metastases[P=0.013,Exp(B)=7.488 ],but not tumor distance to anal verge(P=0.246).The difference in local recurrence rate,overall recurrence rate,disease-free survival rate and overall survival rate between CRMI and that of CRM negative,were significant(P=0.002,0.000,0.000 and 0.003,respectively). Conclusions CRMI is an accurate prognostic factor of recurrence and postoperative survival,and its occurrence is mainly determined by biologic characteristics of the tumor.TME could decrease the incidence of CRMI.

BACKGROUND:Extreme lateral lumbar disc herniation is a rare type of lumbar disc herniation, there are a variety of treatment methods, but the therapeutic efficacy and recurrence rate are controversial.
OBJECTIVE:To investigate the availability of lumbar pedicle screw fixation combined with interbody fusion cage for treating extreme lateral lumbar disc herniation.
METHODWe retrospectively analyzed 19 patients with extreme lateral lumbar disc herniation after treatment with lumbar pedicle screw fixation combining with interbody fusion cage from March 2006 to January 2009. The outcomes were evaluated depending on VAS scoring standard and Macnab scoring standard, lumbar stability were observed postoperatively. We analyzed the spinal stability in recurrent lumbar disc herniation patients after lumbar pedicle screw fixation combined with interbody fusion cage depending on literature search.
RESULTS AND CONCLUSION:Al the 19 patients were fol owed up for 13 months to 3 years, the leg and lumbar pain of al the patients were relieved to varying degrees. Preoperative VAS score was 7.3±1.28 points and postoperative VAS score was 2.1±0.8 points, showing significant difference between two groups (P<0.05). The excellent and good rate was up to 95%with 15 excellent results, 3 good results and 1 acceptable result depending on Macnab evaluation standard. There was no pedicle screw loosening, broken, non-fusion phenomenon. Al the lumbar interbody fusions were good. No one occurred secondary lumbar spinal stenosis. Experimental findings indicate that, lumbar pedicle screw fixation combined with interbody fusion cage for extremely lateral lumbar disc herniation, is characterized as fast symptom relief, strong fixation and good lumbar stability.

OBJECTIVETo investigate the impact of adding folic acid, vitamin B(12) and probucol to standard antihypertensive medication on plasma homocysteine (Hcy) and asymmetric dimethylarginine (ADMA), serum NO and eNOS of essential hypertensive patients.

METHODA total of 120 patients with hypertension were randomly divided to three groups (n = 40 each): group 1 (standard medication), group 2 (adding folic acid 5 mg/day and vitamin B(12) 500 µg twice daily) and group 3 (adding folic acid 5 mg/day and vitamin B(12) 500 µg twice daily and probucol 500 mg twice daily). Plasma Hcy and ADMA, serum NO and eNOS levels were observed at baseline, 2 and 12 weeks after various therapy.

RESULTSIn group 1, concentrations of plasma Hcy [(23.06 ± 14.15) µmol/L, (23.67 ± 12.31) µmol/L, (23.25 ± 11.64) µmol/L], ADMA [(0.21 ± 0.12) µmol/L, (0.23 ± 0.13) µmol/L, (0.21 ± 0.09) µmol/L] and serum NO [(64.14 ± 15.07) µmol/L, (65.29 ± 15.04) µmol/L, (65.32 ± 13.58) µmol/L], eNOS [(20.02 ± 4.50) µg/L, (20.79 ± 4.03) µg/L, (19.82 ± 5.70) µg/L] remained unchanged during the 12 weeks therapy (all P > 0.05). In group 2, concentrations of plasma Hcy [(12.54 ± 6.49) µmol/L] and ADMA[(0.18 ± 0.07) µmol/L] were significantly decreased after the treatment of 12 weeks than the treatment baseline value [(21.51 ± 7.82) µmol/L, (0.20 ± 0.12) µmol/L] and 2 weeks value[(19.38 ± 8.14) µmol/L, (0.21 ± 0.12) µmol/L], however the concentrations of serum NO and eNOS showed contrary results of the Hcy and ADMA's. (all P < 0.05). In group 3, similar changes occurred at 2 weeks after therapy (P < 0.05 2 weeks vs. baseline and 12 weeks vs. 2 weeks). Plasma ADMA level was positively correlated with Hcy at baseline (r = 0.546, P < 0.05).

CONCLUSIONSSupplementation of folic acid, VitB(12) and/or probucol helps to improve endothelial function and reduce plasma Hcy and ADMA levels in patients with hypertension.

Aged ; Antihypertensive Agents ; therapeutic use ; Arginine ; analogs & derivatives ; blood ; Female ; Folic Acid ; therapeutic use ; Homocysteine ; blood ; Humans ; Hypertension ; blood ; drug therapy ; Male ; Middle Aged ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type III ; blood ; Plasma ; metabolism ; Probucol ; therapeutic use ; Vitamin B 12 ; therapeutic use ; Vitamin B Complex ; therapeutic use ; Vitamins ; therapeutic use

Objective To screen serum proteomic marker of hepatic echinococcosis, establish a diagnotic model of serum protein fingerprint patterns, and evaluate its clinical application for hepatic echinococcosis. Methods Serum samples from 68 patients with hepatic echinococcosis matched with 73 controls composed of 33 patients with liver diseases other than hepatic echinococcosis and 40 healthy people were collected. All subjects were divided into training group (37) and testing group (67). Serum protein profiling of patients with hepatic echinococcosis and controls were detected using surface enhanced laser desorption/ionization time of flight mass spectrometry(SELDI-TOF-MS) and weak cation exchange protein chip(WCX2). Peak intensities were compared, in the training group, between 37 patients with hepatic echinococcosis and 37 controls, 5 patients with HCE and 5 patients with HAE, and 8 patients with hepatic echinococcosis before and after operation, respectively. ZJU-Protein Chip Data Analyze System(ZJU-PDAS) was used for data analysis and the model of serum protein fingerprint patterns was build by support vector machine (SVM). The sensitivity and specificity of the model for diagnosis of hepatic echinococcosis were verified by blind method on samples of testing group. Results There were nine different protein peak spectra between hepatic echinococcosis group and control group, of which eight protein peak spectra decreased in patient group, their relative molecular mass were 1044, 1047, 1073, 1075, 1338, 6453, 6649, 8714 m/z, respectively, while one protein peak spectrum(5651 m/z) increased(P < 0.05). The sensitivity,specificity, positive predictive and negative predictive value of the model validated by blind method were 77.4% (24/31), 66.7% (24/36), respectively. There were two different protein peak spectra between HCE group and HAE group, Their relative molecule mass were 8716 and 2751 m/z, respectively (P < 0.05). Six different proteins were detected from pre-operation group and post-operation group. Their relative molecular mass were 1297, 1505, 1525, 1534, 5921, 5941 m/z, respectively(P < 0.05). Conclusions It is a successful way to screen serum proteomic marker in patients with hepatic echinococcosis by SELDI-TOF-MS and Bio-informatics, and the marker has a potential clinical value in diagnosis and judging prognosis of hepatic echinococcosis.

OBJECTIVETo compare the clinical curative effect of thoracolumbar burst fracture treated by the posterior unilateral approach corpectomy fusion screw-rod fixation and anterior corpectomy bone fusion screw plate fixation.

METHODSFrom January 2008 to May 2014,36 cases of thoracolumbar burst fracture underwent operation of decompression, fusion, and internal fixation was retrospective analyzed. Among them, 16 patients were treated through posterior approach as posterior group, including 13 males and 3 females aged from 37 to 62 years old; 9 cases caused by falling injury, 3 cases by traffic accident injury,4 cases by heavy aboved;the injury segment was on T₁₂ in 2 cases, L₁ in 5 cases, L₂ in 7 cases, L₃ in 2 cases; according ASIA grade, 3 cases were grade A, 2 cases were grade B, 2 cases were grade C, 5 cases were grade D, 4 cases were grade E; the time between injury and operation ranged from 5 to 15 days. Other 20 patients were treated through anterior-lateral approach as anterior-lateral group, including 15 males and 5 females with age from 27 to 62 years old; 12 cases caused by falling injury, 4 cases by traffic accident injury, 4 cases by heavy aboved; the injury segment was on T₁₂ in 2 cases, L₁, in 7 cases, L₂ in 9 cases, L₃ in 2 cases; for ASIA grade: 4 cases were grade A, 2 cases were grade B, 4 cases were grade C, 6 cases were grade D, 4 cases were grade E; the time between injury and operation ranged from 4 to 12 days. The operation time, bleeding during operation and postoperative drainage volume were observed in two groups,and the changes of nerve function of ASIA grade, clinical efficacy,improved degree of thoracic and lumbar lordosis,and bony fusion were compared between two groups.

RESULTSAll patients were followed up from 12 to 24 months with an average of (15.8 ± 3.3) months. The operation time, bleeding during operation, and postoperative drainage volume had no significant different between two groups (P > 0.05). As compared with preoperative, ASIA grade of two groups at last follow-up had statistically significantly different (P < 0.01), the neural function of two groups after operation was recovered for different extent. The JOA score of two groups was compared between last follow-up and preoperative, the difference had statistically significant (P < 0.01), the two groups showed good clinical effect. The clinical results of ASIA grade, JOA score and RIS had no significant differences between two groups. All patients of two groups were obtained fusion. Thoracic and lumbar lordosis angle improvement degree had no significant difference between two groups ,it bad significant difference had statistical significance compared with preoperative, the two approaches could effectively restore the spinal sequence.

CONCLUSIONFor patients with thoracolumbar burst fracture just treated by anterior decompression and reconstruction of anterior column, according to the degree of operation performer' skill proficiency and the patient' condition to choose, but for patients must performed the spinal canal decompression anterior and posterior, the three column-reconstruction to required anterior-posterior approach, the posterior unilateral approach corpectomy fusion screw-rod fixation obviously shorten operation time, reduce the operation wound, it is worth the clinical promotion.

Adult ; Decompression, Surgical ; methods ; Female ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Reconstructive Surgical Procedures ; Retrospective Studies ; Spinal Fractures ; surgery ; Spinal Fusion ; methods ; Thoracic Vertebrae ; injuries ; surgery