Objective To investigate the regulatory effects of PTEN-PI3K/AKT pathway on the apoptosis of gastric cancer MKN28 cells and the possible mechanisms.Methods The specific sequence of PTEN was transfected to MKN28 cells by eukaryotic expression vector (transfection group),and then vacant vector (negative control group) and PBS (blank control group) were transfected to the MKN28 cells,respectively.The effects of PTEN-PI3K/AKT pathway on the apoptosis of MKN28 cells and the expressions of PI3K,AKT,Caspase-3 and Caspase-9 were investigated.The growth curve and apoptosis of the MKN28 cells were detected by MTT assay and TUNEL staining,respectively,and the protein expression was detected by the Western blot.MKN28 cells which did not transfected by the PTEN were processed by inhibitor of PI3K (LY294002) (treated group),and MKN28 cells in the control group were processed by PBS.The expressions of apoptosis protein and apoptosis-related protein were detected after inhibition of PI3 K.All data were analyzed using the t test.Results The model of over-expression of PTEN was obtained and transfected into MKN28 cells.The survival of MKN28 cells in the transfection group was significantly inhibited in a time-dependant manner ( r =0.938,P ＜ 0.05 ).The mean apoptotic rate of the transfection rate was 27.86％ ± 4.78％,which was significantly higher than 0.01％ ± 0.01％ of the negative control group ( t =9.527,P ＜ 0.05 ).The protein expression of PI3K in the transfection group was 0.25 ± 0.03,which was significantly lower than 0.93 ± 0.16 of the blank control group and 0.96 ± 0.15 of the negative control group (t =7.235,8.883,P ＜ 0.05 ).The protein expression of P-AKT in the transfection group was 0.21 ± 0.03,which was significantly lower than 0.93 ± 0.13 of the blank control group and 0.91 ± 0.12 of the negative control group (t =9.347,9.802,P ＜ 0.05 ).The expressions of Caspase-3 and Caspase-9 of the transfection group were 0.86 ± 0.11 and 0.87 ± 0.12,which were significantly higher than 0.16 ± 0.03 and 0.18 ± 0.04 of the negative control group,and 0.15 ± 0.02 and 0.16 ± 0.03 of the blank control group ( t =10.634,10.999,9.448,9.942,P ＜0.05).The apoptotic rate of the MKN28 cells of the treated group was 28.60％ ± 4.50％,which was significantly higher than 0.12％ ± 0.06％ of the control group (t =10.961,P ＜ 0.05 ).The protein expression of PI3K and P-AKT of the treated group were 0.18 ± 0.02 and 0.11 ± 0.01,which were significantly lower than 0.93 ± 0.14 and 0.90 ± 0.12 of the control group (t =9.186,11.363,P ＜ 0.05 ).The protein expression of PTEN of the treated group was 1.15 ± 0.15,which was significantly higher than 0.21 ± 0.08 of the control group (t =9.577,P ＜ 0.05 ).The relative expressions of Caspase-3 and Caspase-9 of the treated group were 0.86 ± 0.12 and 0.88 ± 0.11,which were significantly higher than 0.25 ± 0.02 and 0.21 ± 0.03 of the control group (t =8.685,10.178,P ＜ 0.05).Conclusions Over-expression of PTEN may enhance the apoptosis of gastric cancer cells through inhibition of PI3K/AKT pathway.Inhibition of PI3K can enhance the expression of PTEN.PI3K and PTEN regulate the apoptosis of gastric cancer cells.

Objective To observe the long-term effect of surgical treatment in infertile varicocele patients. Methods 180 infertile patients with varicocele accepted modified Palomo operation (retroperitoneal high ligation of internal spermatic cord vein) from 2004 to 2007, and the long-term effect of thesurgery were analyzed retrospectively, such as semen parameters and spouse pregnancy. Results The operations were performed successfully in 180 cases, and semen parameters improved significantly after the surgery (Ps ＜ 0. 05 ).Among all patients, 10 cases had spouse pregnancy within 3 months after operation;47 cases had spouse pregnancy from 3 to 6 months after operation; 25 cases had spouse pregnancy from 6 to 12 months after operation;13 cases had spouse pregnancy from 12 to 24 months after operation. The fertility rate was 52. 8% (95/180) in total. Conclusion Surgical treatment of infertile male patients with varicocele has good long-term effects, made it the first choice in these patients.

Objective To observe and analyze the clinical curative effect of the platelet rich plasma (PRP) in treatment of chronic difficult to heal the wound.Methods We selected 60 patients with chronic difficult healing wounds and they were divide into two groups at random,30 patients of A group,B group of 30 patients.All patients were given routine debridement,drainage and decompression treatment,B group was given autologous PRP wound injection and follow-up on the basis of conventional treatment.After injection of PRP,platelet count,platelet recovery and application of PRP granulation tissue growth and wound healing were observed.Results The wound healing effect at the 7th,14th,21 st day between the two groups,and compared with the 1 st day,the differences were statistically significant (A group:10.28％,22.16％,43.25％,65.78％;B group:18.75％,37.58％,61.84％,80.26％;X2:7.895,8.934,10.231,9.076,all P ＜ 0.05).Two groups of patients had obvious difference in healing time,in B group the average healing time was (12.6 ± 5.2) d,which in was (17.3 ± 7.4) d,the difference was statistically significant (t =1.932,P ＜ 0.05).Conclusion The PRP can effectively promote the wound repair of soft tissue,to promote healing of chronic difficult wound healing,the curative effect is distinct,it is worthy of clinical popularization and application.

Objective To investigate clinical relevance of the acute Stanford B aortic dissection vascular repair time for treatment and prognosis.Methods A retrospective clinical data of our hospital treated 1 10 cases of vascular repair of acute aortic dissection type Stanford B were analyzed according to the onset to treatment time.They were divided into three groups:group A (onset to treatment time of 7 days),group B (onset to treatment time 14) and group C (onset to treatment time than 14 days);the three groups were observed after aortic repair status and prognosis related indicators.Results The true lumen false lumen diameter and maximum diameter ratio in group A was the largest (1.8 ±0.5),which was significantly higher than that of group B and group C (1.3 ±0.3)and (1.3 ± 0.2),the difference was statistically significant (F=7.352,P<0.05).Ratio of preoperative minimum true lumen diameter and the maximum diameter of the false lumen,ratio of postoperative maximum true lumen diameter and the maximum diameter of the false lumen were associated with the onset of the endovascular repair time,showed a negative correlation (r=-0.452,-0.281,all P<0.05).postoperative complication rate of group A was 12.8%,complica-tion rate of group B was 29.5%,complication rate of group C was 33.3%,data of group A was significantly lower than those of group B and group C,the difference were statistically significant (χ2 =4.024,4.721,all P<0.05).group A died three cases,group B had 1 patient died and in group C there was no deaths.Hospital stay and hospital costs of group A were significantly lower than those of group B and group C,the difference werestatistically significant (F=4.125,7.375,all P<0.05).Conclusion As soon as possible to implement acute aortic dissection stanford B aortic vascular repair could better improve outcomes,and reduce the cost of hospitalization and treatment.At the same time, it can reduce the incidence of complications and improve clinical safety.

Objective To investigate the epididymal sperm ultrastructure after vasectomy. Methods During the operation of vasovasostomy (38 cases, test group) and vasectomy (13 cases,control group,group A) the proximal vas deferens fluids were collected.The epididymal sperm morphology in the proximal vas deferens fluids was observed using optical and electronic microscopes.Data were analyzed by SPSS program. Results The data from group A showed that there was no significant difference between left and right side in the percentage of normal sperm morphology.The results from both optical and electronic microscopes showed that there was a significant difference in the percentage of normal sperm morphology (a),abnormal sperm head (b),sperm with neck defect (c) and sperm with tail defect (d) between group A and group B(5 years after vasectomy). (electronic microscopes:(a):(40.28?11.53)% vs (16.80?7.93)%,(6.29? 4.57 )%,( 4.63? 5.06 )%; (b):(35.00?14.18)% vs (59.05?14.44)%,(63.43?15.23)%,(82.05? 16.71 )%;(c):(20.83? 6.40 )% vs (13.60?6.78)%,( 14.71? 6.82)%,(9.00?7.18)%;(d):(3.89?4.44)% vs (10.55?11.73)%,(15.57?9.81)%,(4.32?7.65)%. optical microscopes:(a):(49.12?20.55)% vs (19.95?15.42)%,(10.00?9.50)%,(5.84?9.63)%; (b):(35.00?14.55)% vs (22.55?16.24)%,(14.71?15.78)%,(10.68?18.65)%; (d):(15.80?9.55)% vs (57.50?24.74)%,(75.29?23.90)%,(78.21?30.33)%.There existed multi-cell organ and multi-form abnormalities in the inner structures of sperm heads,necks and tails. Conclusions The epididymal sperm defect (including head, neck and tail) after vasectomy was demonstrated in the inner cell organs.The longer after vasectomy, the higher percentage of sperm with abnormality was found.

The guanine-nucleotide exchange factor (GEF) RalGPS1a activates small GTPase Ral proteins such as RalA and RalB by stimulating the exchange of Ral bound GDP to GTP, thus regulating various downstream cellular processes. RalGPS1a is composed of an Nterminal Cdc25-like catalytic domain, followed by a PXXP motif and a C-terminal pleckstrin homology (PH) domain. The Cdc25 domain of RalGPS1a, which shares about 30% sequence identity with other Cdc25-domain proteins, is thought to be directly engaged in binding and activating the substrate Ral protein. Here we report the crystal structure of the Cdc25 domain of RalGPS1a. The bowl shaped structure is homologous to the Cdc25 domains of SOS and RasGRF1. The most remarkable difference between these three Cdc25 domains lies in their active sites, referred to as the helical hairpin region. Consistent with previous enzymological studies, the helical hairpin of RalGPS1a adopts a conformation favorable for substrate binding. A modeled RalGPS1a-RalA complex structure reveals an extensive binding surface similar to that of the SOS-Ras complex. However, analysis of the electrostatic surface potential suggests an interaction mode between the RalGPS1a active site helical hairpin and the switch 1 region of substrate RalA distinct from that of the SOS-Ras complex.
Amino Acid Sequence ; Binding Sites ; Catalytic Domain ; Cloning, Molecular ; Crystallography, X-Ray ; Escherichia coli ; Guanosine Diphosphate ; metabolism ; Guanosine Triphosphate ; metabolism ; Humans ; Models, Molecular ; Molecular Conformation ; Molecular Sequence Data ; Plasmids ; metabolism ; Protein Binding ; Protein Structure, Tertiary ; genetics ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; ral GTP-Binding Proteins ; chemistry ; genetics ; metabolism ; ral Guanine Nucleotide Exchange Factor ; chemistry ; genetics ; metabolism

OBJECTIVETo observe sperm motion parameters in pre-freeze and post-thaw semen samples using computer-assisted sperm analysis (CASA) system.

METHODSSemen analyses of 238 samples before freezing and after thawing were separately performed by Hamilton-Thorne Sperm Analyzer.

RESULTSSperm motility in post-thaw samples was significantly decreased. There was significant correlation and difference between pre-freeze and post-thaw samples in sperm motion parameters, including average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), straightness (STR) and linearity (LIN), except heat cross frequency (BCF). The percentage of sperm movement velocity parameters (VAP, VSL and VCL) and moving pattern parameters (ALH) significantly decreased, while that of LIN and STR significantly increased in post-thaw samples.

CONCLUSIONCASA system is of clinically applied value and is a useful tool for evaluating sperm motion parameters in pre-freeze and post-thaw semen samples.

Autoimmune epilepsy (AE) is a type of epilepsy mediated by autoantibodies and immune cells. The main pathogenesis of AE is that autoimmune antibodies related to AE interact with surface of nervous cell membrane antigens or intracellular antigens, which leads to the disorder of excitatory and inhibitory nervous system and causes epileptic seizures. Glutamate receptor subunit 3 peptide B antibodies (GluR3B Ab's) are one type of anti-neuron autoantibodies related to AE, and they can combine with GluR3B subunit of α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor (AMPAR), which causes an increase of intracellular calcium. Intracellular calcium overload further leads to dysregulation of energy metabolism in neurons or oligodendrocyte progenitor cells (OPCs). The damage of neurons or OPCs eventually trigger seizure. Here, we mainly discuss the mechanisms of GluR3B Ab's involving in the development of autoimmune epilepsy.

Objective: This study’s objective is to assess the efficacy and safety of Pulsed Magnetic Therapy System (PMTS) in improving insomnia disorder. Methods: Participants with insomnia disorder were randomly assigned to receive either PMTS or sham treatment for four weeks (n= 153; PMTS: 76, sham: 77). Primary outcomes are the Insomnia Severity Index (ISI) scores at week 0 (baseline), 1, 2, 3, 4 (treatment), and 5 (follow-up). Secondary outcomes are the Pittsburgh Sleep Quality Index at baseline and week 4, and weekly sleep diary-derived values for sleep latency, sleep efficiency, real sleep time, waking after sleep onset, and sleep duration. Results: The ISI scores of the PMTS group and the sham group were 7.13±0.50, 11.07±0.51 at week 4, respectively. There was a significant group×time interaction for ISI (F3.214, 485.271=24.25, p<0.001, ηp 2=0.138). Only the PMTS group experienced continuous improvement throughout the study; in contrast, the sham group only experienced a modest improvement after the first week of therapy. At the end of the treatment and one week after it, the response of the PMTS group were 69.7% (95% confidence interval [CI]: 58.6%–79.0%), 75.0% (95% CI: 64.1%–83.4%), respectively, which were higher than the response of the sham group (p<0.001). For each of the secondary outcomes, similar group×time interactions were discovered. The effects of the treatment persisted for at least a week. Conclusion PMTS is safe and effective in improving insomnia disorders.

Objective: To investigate the efficacy of posterior decompression pedicle screw fixation and single pedicle screw fixation for thoracolumbar fracture with greenstick lamina fracture. Methods: A retrospective case series study was conducted to analyze the clinical data of 106 patients with thoracolumbar fractures combined with greenstick lamina fractures admitted to the 909th hospital from January 2011 to May 2016. There were 68 males and 38 females, aged 21-58 years [(39.5±7.1)years]. The fracture was located at T₁₁ in 9 patients, at T₁₂ in 6, at L₁ in 28, at L₃ in 11. A total of 58 patients underwent posterior decompression pedicle screw fixation including 37 males and 21 females, aged 23-58 years [(38.7±6.6)years]. The preoperative neurological function was evaluated according to ASIA grade: grade A for three patients, grade B for nine, grade C for 14, grade D for 27, and grade E for 53 patients. A total of 48 patients were treated with single posterior pedicle screws fixation including 31 males and 17 females, aged 21-57 years [(41.4±5.8)years]. Statistical indicators included operation time, intraoperative blood loss, anterior height ratio of injured vertebrae, sagittal Cobb angle, visual analogue scale (VAS), ASIA grading, dural tears and/or cauda equina entrapment, and complications. Results: All patients were followed up for 24-72 months [(30.2±4.7)months]. The operation time ranged from 105 to 137 minutes [(113.5±21.3)minutes], and the intraoperative blood loss was 235-310 ml [(252.2±28.6)ml] in the posterior decompression and pedicle screws fixation group. In the posterior pedicle screw fixation group, the operation time ranged from 52 to 85 minutes [(65.3±9.6)minutes], and the intraoperative blood loss was 72-125 ml [(90.2±23.6)ml]. The anterior height ratio of injured vertebrae, sagittal Cobb angle and VAS score of the two groups were significantly improved immediately after operation and at the last follow-up (all P<0.01). At one year follow up, the ASIA grading in patients treated with single posterior pedicle screw fixation remained the same while the ASIA grading of patients treated with posterior decompression pedicle screw fixation was significantly improved compared with before operation. Dural tears were found in 12 patients and cauda equina entrapment were found in five patients during decompression. Three patients had neurological function injury after posterior pedicle screw internal fixation surgery, and they received a second decompression operation. Dural tears and/or cauda equina entrapment were subsequently found in all of the three patients. No complications related to internal fixation occurred during follow-up. Conclusions Both posterior decompression pedicle screw fixation and single pedicle screw fixation can effectively correct the kyphosis deformity, restore vertebral body height, and relieve pain in the treatment of thoracolumbar fracture with greenstick lamina fracture. Dural tears and/or cauda equina entrapment might be subsequently found in greenstick lamina fracture. Posterior pedicle screw fixation can not release the entrapped cauda nerve, so attention should be paid to the risk of secondary surgery.