BACKGROUND:The state of ischemic penumbra after cerebral infraction determines the final infarct volume. Decreased selective gene expression and protein synthesis, lactic acidosis and cytotoxic edema, salvage therapy of nerve cels, and continous regulation of neurovascular unit in the ischemic penumbra are areas of interest for recovery of neurological function. OBJECTIVE: Taking the neoangiogenesis process of post-infarction as the research point, to investigate the basic law of neovascularization with time and the effect of electroacupuncture intervention on proliferation of local blood vessels. METHODS: Ninety-six Wistar rats were randomly and evenly divided into model and electroacupuncture groups. The rat models of middle cerebral artery infarction were established by suture method. Immediately after modeling, the rats in the electroacupuncture group were given electrical stimulation (15 Hz, 1mA, 20 minutes) at the Renzhong (Du26) acupoint. The rats in the model group received similar procedures with the exception of no treatment. The angiogenesis was observed using double-immunofluorescence labeling for von Wilebrand Factor (vWF) and Ki-67 antibodies at 3, 6, 12, 24, 48 hours and 3, 7, 12 days after middle cerebral artery occlusion. RESULTS AND CONCLUSION:In the model group, vascular endothelial cel proliferation in the peri-infarct region was not observed at 3, 6 and 12 hours, occurred at 24 hours, increased at 48 hours, reached a maximum at 3 days, began to decrease at 7 days, and disappeared at 12 days after middle cerebral artery occlusion. In the electroacupuncture group, vascular endothelial cel proliferation in the peri-infarct region was not observed at 3 and 6 hours, occurred at 12 hours, increased at 24 and 48 hours, peaked at 3 days, began to decerase at 7 days, and disappeared at 12 days after middle cerebral artery occlusion. Compared with the model group, the time at which vascular endothelial cel proliferation started was earlier, and the number of proliferated vascular endothelial cels was greater in the electroacupuncture group. Vascular endothelial cel proliferation in the infarcted regions and contralateral hemisphere was not observed in either group at al designated time points. These results demonstrate that electroacupuncture intervention can improve the prognosis of cerebral infarction by promoting the proliferation of vascular endothelial cels in the peri-infarct region and advancing the time at which vascular endothelial cel proliferation started in rat models of middle cerebral artery occlusion.

AIM: To explore the molecular mechanism of asthenospermia(AST) by preliminary screening of nucleotide sequences from the ND3 and ND4L genes of mitochondrial DNA(mtDNA). METHODS: Samples from 50 AST patients and 42 age-matched normal controls were collected according to the WHO criteria. Density gradient centrifugation was applied to separate spermatozoa with different vigor. The ND3 and ND4L genes of mtDNA were amplified and sequenced directly from the extracted genomic DNA from AST patients and normal controls. The sequences were compared with revised Cambridge Reference Sequence(rCRS) to analyze the variants. RESULTS: A total of 22 nucleotide variations were found in ND3 and ND4L genes of mtDNA in asthenospermia group and control group. G10320A, A10398G and T10609C were missense mutations, while A10157G and A10313C were the reported for the first time in this study. Haplotype N in patients with AST(33/50) was higher than that in control group(14/42, P<0.05), and haplotype R9 in patients with AST(15/50) was also higher than that in control group(4/42, P<0.05) through genetic testing of ND3 gene. Rates of sperm progressive motility of haplotype F1, F2 and R9 were significantly lower than those of haplotype M and M rest. Two haplotype differences, haplotype M and N, were found in the same AST patient's spermatozoas which had different vigor. Haplotype M had stronger vigor, while haplotype N had lower vigor. By sequencing ND3 gene of mtDNA from 50 AST patients, we detected G10310A heteroplasmic mutation in 2 specimens of asthenospermia with poor and moderate motility spermatozoa, respectively. No mutation occurred in good motility spermatozoa. CONCLUSION: Haplotype of mitochondrial may have some correlation with sperm motility. The nt10398G-10400T polymorphisms may have benefit for sperm motility, whereas the mutation in nt10310A may impair sperm motility.

Objective To investigate the relationship between mtND4 point mutation in sperms and asthenospermia. Methods Fifty-six asthenospermia cases and 44 control cases were collected using the WHO criterion for defining asthenospermia, the regions of mtND4 gene were amplified by using PCR of 3 pairs primers. Consequently, the point mutation, missense mutation and multiple single nucleotide polymorphisms (SNP) were analyzed by employing sequencing technology and bioinformatics tools. Results Six mutations never before identified were found. The frequency of single point mutation T10873C and T11944C in the control group were significantly higher than those in the asthenospermia group (P＜0.05). Eight cases involved T10873C or T11944C among the 10 cases in control groups with missense mutations were found. But, there were only 2 cases with such mutation in the 10 asthenospermia cases with missense mutations (P＜0.05). The previous 20 cases of missense mutations can be described as either multiple SNP group (with T10873C or T11944C) or nonmultiple SNP group. The percentage of a range and a plus b range of multiple SNP group of sperm was significantly higher than the non-multiple SNP group(P＜0.05). Conclusions mtND4 gene mutation, especially the missense mutation may induce loss of sperm motility. The mutations of T10873C and T11944C may be useful for sperm motility or counteract the influence for the sperm motility caused by these harmful mutations.

BACKGROUND:Knee osteoarthritis is a chronic disease characterized as cartilage degeneration and hyperostosis to impact the soft tissues around the joints, thereby resulting in joint pain, and it can be aggravated and lead to joint deformity after weight-bearing. OBJECTIVE:On the basis of imaging parameters of knee osteoarthritis patients and normal controls, a model was established. And then, knee squatting movement was simulated on the model and stress data from the cartilage surface of patients and normal controls were colected and compared, thereby to obtain the stress characteristics of the cartilage surface in knee osteoarthritis patients. METHODS: There were 30 knee osteoarthritis patients and 30 healthy volunteers in the study. CT and MRI data from these participants were colected and analyzed using Mimics software and simpleware software to establish a model. Based on this model, cartilage-related stress data were harvested from knee osteoarthritis patients and healthy volunteers and compared to summarize the stress characteristics of the articular cartilage in patients with knee osteoarthritis. RESULTS AND CONCLUSION:In knee osteoarthritis patients, the stress on the cartilage surface from standing to squatting was increased in a nonlinear manner. During the process of squatting, the mediolateral stress was higher in the knee osteoarthritis patients than healthy controls (P < 0.05); however, there was no difference in the proximodistal and anteroposterior cartilage surface stress between the two groups (P > 0.05). These findings indicate that the knee osteoarthritis musculoskeletal model and finite element model are established successfuly, and these models are closer to the real motion characteristics of the knee joints, based on which, quantitative biomechanical data of the knee can be provided for knee osteoarthritis treatment.

Objective This study was aimed to investigate the effects of ω-3 polyunstaurated fatty acids (ω-3 PUFAs) on the growth of gastric cancer cells in nude mice,and to find whether the Ros homolog gene Rho-associated coiled-coil containing protein kinase 1 (RHO-ROCK1) signaling pathway is involved.Methods 16 BALB/C nude mice were injected subcutaneously with SGC7901 gastric cancer cells to establish the tumor-bearing mouse model.The mice were randomized:control group (normal saline) and intervention group (ω-3 PUFAs).The mRNA expression of Ros homolog gene family,member A (RHOA),RHOC,and ROCK1 in tumor tissue were detected by quantitative polymerase chain reaction (qPCR).Immunofluorescence and Western blot were used to detect RHOA,RHOC,and ROCK1 protein expression.Results The volume and weight of the tumors in the ω-3 PUFAs group were slightly smaller than that in the control group (P ＞ 0.05).Compared to the control group,hematoxylin and eosin staining showed multifocal tumor necrosis in the ω-3 PUFAs group,while the tumors of the control group showed abundant blood supply.qPCR and Western blot showed that the mRNA and proteins expression of RHOA and ROCK1 in the ω-3 PUFAs group was significantly lower than those in the control group (P ＜ 0.05).The immunofluorescence redults also showed that the expression of these proteins in the ω-3 PUFAs group was slightly lower than that in the control group.Conclusions These results suggested that ω-3 PUFAs may affect the growth of gastric cancer in nude mice by affecting the expression of RHOA,RHOC and ROCK1,thus inhibiting the excessive proliferation of gastric cancer cells and leading to tumor necrosis.

Objective To study the effect of the preset temperature,heating time and the spacing of radio frequency electrodes.Methods Fresh egg white was used to study the effects of heater spacing,temperature and heating time.Two electrodes were fixed to keep the needles perpendicular to the center of egg white confined on a square plate.Temperatures from 50 to 90℃ were tested with heating times of 60 and 120 seconds and electrode spac ings of 2.0,1.0,0.5 and 0.25 centimetres.The real-time temperature and the time to the appearance of a spindleshaped bridge between the electrodes was observed.The egg white's impedance and resistance,the lowest temperature of denaturation,and the mass (or volume) denatured at different time points were also observed.Results With an electrode spacing of 2 cm and the thermostat set at 63℃,the true electrode temperatures were 63.7 and 52.6℃,with the secondary electrode the cooler.Significant differences between the secondary electrode temperature and the preset temperature as well as the temperature between the primary and the secondary electrodes were observed.It was also found that different electrode spacing caused significant differences between the preset temperature and the primary as well as the secondary electrode temperature.Denaturation began at 56℃ with a spacing of 0.25 cm.At spacings of 0.25 and 0.50 cm,a spindle-shaped connection between the two electrodes was observed at temperatures above 70℃,but higher temperature was required at 1.00 and 2.00 cm.When the spacing was 0.25 cm and the preset temperature was 75℃,the denatured volume after 60 s of heating was 90.21+0.64 mm3.Heating for 120 s denatured 95.08+ 0.53 mm3 and two 60.s heatings 92.88+0.74 mm3,all significant differences.There were no significant differences in the resistance of the egg white before and after a single heating for 60 s,but after a single heating lasting 120 s it had increased significantly from 128.41+8.04 to 121.29±8.04 Ω.Conclusion Bipolar radio frequency heating can denature egg white.Higher temperature,longer heating and smaller electrode spacing heat more effectively.