Magnetic resonance diffusion tensor imaging following major ozonated autohemotherapy for treatment of acute cerebral infarction

Major ozonated autohemotherapy has been shown to promote recovery of upper limb motor function in patients with acute cerebral infarction, but whether naajor ozonated autohelnotherapy affects remote in）ury remains poorly understood. Here, we assumed that major ozonated autohemotherapy contributes to recovery of clinical function, possibly by reducing remote injury after acute cerebral infarction. Sixty acute cerebral infarction patients aged 30-80 years were equally and randomly allocated to ozone treatment and control groups. Patients in the ozone treatment group received medical treatment and major ozonated autohemotherapy （47 mg/L, 100 mL ozone） for 10 ± 2 days. Patients in the control group received medical treatment only. National Institutes of Health Stroke Scale score, modified Rankin scale score, and reduced degree of fractional anisotropy values of brain magnetic resonance diffusion tensor imaging were remarkably decreased, brain function improved, clinical efficiency significantly increased, and no obvious adverse reactions detected in the ozone treatment group compared with the control group. These findings suggest that major ozonated autohemotherapy promotes recovery of neurological function in acute cerebral infarction patients by reducing re,note injury, and additionally, exhibits high safety.

Major ozonated autohemotherapy promotes the recovery of upper limb motor function in patients with acute cerebral infarction

Major ozonated autohemotherapy is classically used in treating ischemic disorder of the lower limbs In the present study, we performed major ozonated autohemotherapy treatment in patients with acute cerebral infarction, and assessed outcomes according to the U.S. National Institutes of Health Stroke Score, Modified Rankin Scale, and transcranial magnetic stimulation motor-evoked potential. Compared with the control group, the clinical total effective rate and the cortical potential rise rate of the upper limbs were significantly higher, the central motor conduction time of upper limb was significantly shorter, and the upper limb motor-evoked potential amplitude was significantly increased, in the ozone group. In the ozone group, the National Institutes of Health Stroke Score was positively correlated with the central motor conduction time and the motor-evoked potential amplitude of the upper limb. Central motor conduction time and motor-evoked potential amplitude of the upper limb may be effective indicators of motor-evoked potentials to assess upper limb motor function in cerebral infarct patients. Furthermore, major ozonated autohemotherapy may promote motor function recovery of the upper limb in patients with acute cerebral infarction.

Expression and significance of VEGF and p53 in degenerate intervertebral disc tissue

Objective:To investigate the mechanism of expression and significance of vascular endothelial growth factor(VEGF) and p53 in degenerate intervertebral disc tissue.Methods:Pathological sections collected from 156 patienls with lumbar disc herniation after surgery were tested by immunohistochemistry method,for evaluation of the expression of VEGF and p53 in degenerate intervertebral disc tissue.Results:98 cases(62.8%) with vascular infiltration phenomenon are found,and positive rates of VEGF and p53 in degenerate intervertebral disc tissue are 73.42% (116/156) and 58.97%(92/156);co-expression rate is 53.2%(83/l56);the expression rates of VEFG and p53 are significantly higher in the tissue with blood vessel infiltration than in the tissue without infiltration;there is a close relationship of VEGF with p53.Conclusions:VEGF and p53 gene synergetic express in degenerate intervertebral disc tissue,working together in neovascularization and infiltration,and accelerating intervertebral disc tissue degeneration.

Evaluation of Tubal Patency with Transvaginal Three-dimensional Hysterosalpingo-contrast Sonography

Objective To investigate diagnostic efficacy of transvaginal three-dimensional hysterosalpingo-contrast sonography(3D-Hy Co Sy) in assessing tubal patency with chromolaporoscopy. Methods A total of 157 infertile women underwent 3D-Hy Co Sy to evaluate tubal patency. Among these patients, 39 patients were also examined by chromolaporoscopy. The concordance of the two clinical assessment methods was analyzed by the Kappa coefficient test. Results Among the 306 oviducts examined by 3D-Hy Co Sy, 99(32.4%) were patent, 126(41.2%) partially obstructed, and 81(26.5%) completely obstructed. Diagnostic results with 3D-Hy Co Sy were not statistically different from those obtained in the 39 women(78 oviducts) who also underwent chromolaporoscopy, and the two methods showed a high concordance(κ=0.747, P=0.000). The 3D-Hy Co Sy procedure had a sensitivity of 84.8%(28/33), a specificity of 96.2%(25/26), and positive and negative predictive values of 93.3%(28/30) and 86.2%(25/29) respectively. Conclusion Transvaginal 3D-Hy Co Sy can accurately reveal the spatial path and morphology of the oviduct and is a safe and effective method to evaluate tubal patency.

Inhibition of endoplasmic reticulum stress alleviates secondary injury after traumatic brain injury

Apoptosis after traumatic brain injury has been shown to be a major factor influencing prognosis and outcome. Endoplasmic reticulum stress may be involved in mitochondrial mediated neuronal apoptosis. Therefore, endoplasmic reticulum stress has become an important mechanism of secondary injury after traumatic brain injury. In this study, a rat model of traumatic brain injury was established by lateral fluid percussion injury. Fluorescence assays were used to measure reactive oxygen species content in the cerebral cortex. Western blot assays were used to determine expression of endoplasmic reticulum stress-related proteins. Hematoxylin-eosin staining was used to detect pathological changes in the cerebral cortex. Transmission electron microscopy was used to measure ultrastructural changes in the endoplasmic reticulum and mitochondria. Our results showed activation of the endoplasmic reticulum stress-related unfolded protein response. Meanwhile, both the endoplasmic reticulum stress response and mitochondrial apoptotic pathway were activated at different stages post-traumatic brain injury. Furthermore, pretreatment with the endoplasmic reticulum stress inhibitor, salubrinal（1 mg/kg）, by intraperitoneal injection 30 minutes before injury significantly inhibited the endoplasmic reticulum stress response and reduced apoptosis. Moreover, salubrinal promoted recovery of mitochondrial function and inhibited activation of the mitochondrial apoptotic pathway post-traumatic brain injury. These results suggest that endoplasmic reticulum stress might be a key factor for secondary brain injury post-traumatic brain injury.

Remodeling of motor cortex function in acute cerebral infarction patients following human urinary kallidinogenase A functional magnetic resonance imaging evaluation after 6 months

A total of 29 patients were treated within 48 hours after acute subcortical cerebral infarction with Xuesaitong or Xuesaitong plus human urinary kallidinogenase for 14 days. Neurological deficits, activity of daily living, and evaluations of distal upper limb motor functions at the 6-month follow-up showed that patients treated with Xuesaitong plus human urinary kallidinogenase recovered better than with Xuesaitong alone. In addition, functional MRI revealed that activation sites were primarily at the ipsilesional side of injury in all patients. Human urinary kallidinogenase induced hyperactivation of the ipsilesional primary sensorimotor cortex, premotor cortex, supplementary motor area, and contralesional posterior parietal cortex. Results showed that human urinary kallidinogenase improved symptoms of neurological deficiency by enhancing remodeling of long-term cortical motor function in patients with acute cerebral infarction.

Decreased osteogenesis of adult mesenchymal stem cel s by reactive oxygen species under cyclic stretch: a possible mechanism of age related osteoporosis

Age related defect of the osteogenic differentiation of mesenchymal stem cells（MSCs） plays a key role in osteoporosis. Mechanical loading is one of the most important physical stimuli for osteoblast differentiation.Here, we compared the osteogenic potential of MSCs from young and adult rats under three rounds of 2 h of cyclic stretch of 2.5% elongation at 1 Hz on 3 consecutive days. Cyclic stretch induced a significant osteogenic differentiation of MSCs from young rats, while a compromised osteogenesis in MSCs from the adult rats.Accordingly, there were much more reactive oxygen species（ROS） production in adult MSCs under cyclic stretch compared to young MSCs. Moreover, ROS scavenger N-acetylcysteine rescued the osteogenic differentiation of adult MSCs under cyclic stretch. Gene expression analysis revealed that superoxide dismutase 1（SOD1） was significantly downregulated in those MSCs from adult rats. In summary, our data suggest that reduced SOD1 may result in excessive ROS production in adult MSCs under cyclic stretch, and thus manipulation of the MSCs from the adult donors with antioxidant would improve their osteogenic ability.

Application of restriction display PCR technique in the preparation of cDNA microarray probes

AIM: To develop a simplified and efficient method for the preparation of hepatitis C virus (HCV) cDNA microarray probes.METHODS: With the technique of restriction display PCR (RD-PCR), restriction enzyme Sau3A I was chosen to digest the full-length HCV cDNAs. The products were classified and re-amplified by RD-PCR. We separated the differential genes by polyacrylamide gel electrophoresis and silver staining. Single bands cut out from the polyacrylamide gel were isolated. The third-round PCR was performed using the single bands as PCR template.The RD-PCR fragments were purified and cloned into the pMD18-T vector. The recombinant plasmids were extracted from positive clones, and the target gene fragments were sequenced. The cDNA microarray was prepared by spotting RD-PCR products to the surface of amino-modified glass slides using a robot. We validated the detection of microarray by hybridization and sequence analysis.RESULTS: A total of 24 different cDNA fragments ranging from 200 to 800 bp were isolated and sequenced,which were the specific gene fragments of HCV. These fragments could be further used as probes in microarray preparation. The diagnostic capability of the microarray was evaluated after the washing and scanning steps. The results of hybridization and sequence analysis showed that the specificity, sensitivity, accuracy, reproducibility,and linearity in detecting HCV RNA were satisfactory.CONCLUSION: The RD-PCR technique is of great value in obtaining a large number of size-comparable gene probes, which provides a speedy protocol in generating probes for the preparation of microarrays. Microarray prepared as such could be further optimized and applied in the clinical diagnosis of HCV.

Dysregulation of the miR-146a-Smad4 axis impairs osteogenesis of bone mesenchymal stem cells under inflammation

Osteoporosis is a common disease that affects patient quality of life, especially among the elderly population. Although inflammation contributes significantly to osteoporosis, the underlying mechanism is unclear. In this study, we found that tumor necrosis factor （TNF）-a, an inflammatory environment mimic, inhibits osteogenesis of bone mesenchymal stem ceils （BMSCs）, induces miR-146a and decreases Smad4. Moreover, overexpression of miR-146a inhibited the osteogenic ability of BMSCs, whereas blocking miR-146a partially rescued the osteogenesis deficiency under TNF-a treatment. Molecularly, miR-146a decreased Smad4 expression at the protein level by binding to an element located in the Smad4 3＇-untranslated region, and restoration of Smad4 reversed the inhibitory effects of miR-146a on osteogenesis. Together, our results showed that the inflammatory environment mimic TNF-ol inhibits osteogenesis via upregulation of miR-146a and subsequent downregulation of Smad4, thus suggesting that therapeutic manipulation of miR-146a maybe a potential strategy to improve osteogenesis in the context of osteoporosis.

Association between SNP rs10569304 on the Second Expressed Region of Hole Gene and the Congenital Heart Disease

The correlation of single nucleotide polymorphism (SNP) rs10569304 on the second expressed region of hole gene and congenital heart disease (CHD) of human being, and the effect of hole gene on CHD were investigated. 179 patients with CHD as CHD group and 183 healthy people as control group were selected in the case-control study. DNA was abstracted from the peripheral blood by phenol-chloroform method. Primer was designed for the flanking sequence of SNP rs10569304 on the second expressed region of hole gene. The genotype was identified by PCR degenerative acrylamide electrophoresis with amplification products. Then the three amplification products received sequencing. By chi-square test, the genotype frequency and allele frequency in CHD group and control group were analyzed. There was insertion-deletion (GCC/-) of SNP rs10569304 which corresponded to alleles of A and B in Southern Chinese people. The genotype frequency and allele frequency in control group and CHD group were met the Hardy-Weinberg equilibrium. By chi-square test, in control group and CHD group, the genotype frequency of AA (insertion homozygous), AB (insertion-deletion heterozygous) and BB (deletion homozygous) was 21.31%, 54.09%, 24.59% and 16.75%, 46.36%, 36.87%, respectively. The distributional difference of genotype frequency had statistical significance (χ2=6.51, P<0.05); The allele frequency of A and B was 48.36% and 51.64% in control group, 39.94% and 60.06% in CHD group, respectively. The distribu- tional difference of allele frequency had statistical significance (χ2=5.20, P<0.05). Meanwhile, by contrast with the control group, the BB genotype frequency and B allele frequency in CHD group was higher, but the AA and AB frequency was lower. There was higher risk to suffer from CHD involving B allele. BB genotype had 1.907-fold increased risk of developing CHD according to AA genotype (P<0.05). It is concluded that there is insertion-deletion (GCC/-) of SNP rs10569304 in the Southern Chinese people, and the people whose hole gene involving BB genotype have higher risk to suffering from CHD.

Comparison of TGF-β_1 in tears and corneal haze following Epi-LASIK with and without mitomycin C

AIM: To compare transforming growth factor-β1 (TGF- β1 ) levels in tears and the degree of corneal haze formation following epithelial laser in situ keratomileusis (Epi -LASIK) with and without the use of mitomycin C (MMC) and to investigate the effect of MMC on corneal wound healing. ·METHODS: Thirty-two patients (64 eyes) with high myopia underwent Epi -LASIK surgery, and MMC was randomly used in one eye in each patient. The epithelialization process was observed, and the TGF -β1 level in tears was measured at 1 day, 3, and 7 days postoperatively for comparison with baseline. Corneal haze was graded at 1 month, 3, and 6 months after surgery. · RESULTS: Mean preoperative spherical equivalent refraction was -8.24 ±2.18D (range-6.00 to -10.50D) in the MMC group and -7.82±1.55D (range -6.00 to -9.75D) in the non -MMC group. There was no significant difference between the two groups (P =0.38). Mean epithelialization time was (5.02 ±0.68) days in the MMC group and (4.86±0.57) days in the non-MMC group (P = 0.31). Tear fluid TGF-β1 levels were similar before surgery (P =0.34), but were significantly higher in the non-MMC group at 1 day, 3, and 7 days postoperatively (P =0.004, 0.008, and 0.012, respectively). Corneal haze scores 1 month after surgery were significantly higher in the non -MMC group (P =0.03), and similar at 3 and 6 months after surgery (P =0.28 and 0.62, respectively). ·CONCLUSION: MMC did not delay epithelialization. Inearly postoperative period, lower TGF -β1 levels in tears and a lower grade of corneal haze were observed in the MMC group. Our findings suggest that the ability of MMC to inhibit Epi -LASIK -induced haze might be mediated through TGF-β1 suppression.

Design and Manufacture of Bionic Porous Titanium Alloy Spinal Implant Based on Selective Laser Melting(SLM)

In order to meet the clinical requirements of spine surgery,this paper proposed the exploratory research of computer-aided design and selective laser melting(SLM)fabrication of a bionic porous titanium spine implant.The structural design of the spinal implant is based on CT scanning data to ensure correct matching,and the mechanical properties of the implant are verified by simulation analysis and laser selective melting experiment.The surface roughness of the spinal implant manufactured by SLM without post-processing is Ra 15μm,and the implant is precisely jointed with the photosensitive resin model of the upper and lower spine.The surface micro-hardness of the implant is HV 373,tensile strengthσ_(b)=1238.7 MPa,yield strengthσ_(0.2)=1043.9 MPa,the elongation is 6.43%,and the compressive strength of porous structure under 84.60%porosity is 184.09 MPa,which can meet the requirements of the reconstruction of stable spines.Compared with the traditional implant and intervertebral fusion cage,the bionic porous spinal implant has the advantages of accurate fit,porous bionic structure and recovery of patients,and the ion release experiment proved that implants manufactured by SLM are more suitable for clinical application after certain treatments.The elastic modulus of the sample is improved after heat treatment,mainly because the microstructure of the sample changes fromα’phase toα+βdual-phase after heat treatment.In addition,the design of high-quality bionic porous spinal implants still needs to be optimized for the actual needs of doctors.

Recovery of sympathetic nerve function after lumbar sympathectomy is slower in the hind limbs than in the torso

Local sympathetic denervation by surgical sympathectomy is used in the treatment of lower limb ulcers and ischemia,but the restoration of cutaneous sympathetic nerve functions is less clear.This study aims to explore the recovery of cutaneous sympathetic functions after bilateral L2-4 sympathectomy.The skin temperature of the left feet,using a point monitoring thermometer,increased intraoperatively after sympathectomy.The cytoplasm of sympathetic neurons contained tyrosine hydroxylase and dopamineβ-hydroxylase,visualized by immunofluorescence,indicated the accuracy of sympathectomy.Iodine starch test results suggested that the sweating function of the hind feet plantar skin decreased 2 and 7 weeks after lumbar sympathectomy but had recovered by 3 months.Immunofluorescence and western blot assay results revealed that norepinephrine and dopamineβ-hydroxylase expression in the skin from the sacrococcygeal region and hind feet decreased in the sympathectomized group at 2 weeks.Transmission electron microscopy results showed that perinuclear space and axon demyelination in sympathetic cells in the L5 sympathetic trunks were found in the sympathectomized group 3 months after sympathectomy.Although sympathetic denervation occurred in the sacrococcygeal region and hind feet skin 2 weeks after lumbar sympathectomy,the skin functions recovered gradually over 7 weeks to 3 months.In conclusion,sympathetic functional recovery may account for the recurrence of hyperhidrosis after sympathectomy and the normalization of sympathetic nerve trunks after incomplete injury.The recovery of sympathetic nerve function was slower in the limbs than in the torso after bilateral L（2-4） sympathectomy.

Water-soluble lipopolymer delivery of N-methyl-D-aspartic acid receptor 2B siRNA relieves chronic neuropathic pain in rats

Spinal dorsal horn N-Methyl-D-aspartic acid receptor 2B （NR2B） overexpression plays an important role in the production and maintenance of neuropathic pain. Because small interfering RNA （siRNA） can inhibit NR2B expression, siRNA may provide a novel approach to treat neuropathic pain and possibly nerve injury. However, an efficient and safe vector for NR2B siRNA has not been discovered. This study shows that a water soluble lipopolymer （WSLP） comprised of low molecular weight polyethyleneimine （PEI） and cholesterol can deliver siRNA targeting NR2B for the treatment of neuropathic pain. Results show that intrathecal injection of WSLP/siRNA complexes for 3 days inhibit NR2B gene expression with reductions in mRNA and protein levels by 59% and 54%, respectively, compared with control rats （P 〈 0.01）. Injection of WSLP complexed with scrambled siRNA, or PEI with siRNA did not show this inhibitory effect. Moreover, injection of WSLP/siRNA complexes significantly relieved neuropathic pain at 3, 7, 12, and 21 days, while injection of WSLP with scrambled siRNA or PEI with siRNA did not. These results demonstrate that WSLP can efficiently deliver siRNA targeting NR2B in vivo and relieve neuropathic pain.

Auto-cortex of crystalline lens-induced iris neovascularization

AIM:To investigate auto-cortex of crystalline lens induced iris neovascularization (INV). METHODS: Thirty-six eyes of 36 guinea-pigs were included and divided into three groups randomly in this cohort study. Group A: the right lens nucleus was extracted and the remaining cortical lens material was aspirated thoroughly.. Group B: the lens was removed and 30 mu L precipitated lens cortex was injected into the anterior chamber again. Group C: aspirated the lens cortex of the left eyes and inject them into the right anterior chambers about 10 mu L. Clinical changes were followed by slit-lamp examination and photograph. The eye balls were enucleated at the day of 2, 4, 7, 11, 13, 17 after operation. HE was used to detect the pathological changes. ' RESULTS:Group A:INV had not been observed until the end of empirical study. The stromal layer contained thick wall vessels, without expansion. Group B: All eyes developed INV. Postoperative (po) 7 days; the eyes developed intense and extensive INV. The vessels of iris expanded remarkably and neovascularization was observed erupting from it's lateral wall and stretching towards the anterior surface. Poll days, INV regressed gradually after lens cortex had been absorbed. Group C: Po four (4) days, new blood vessels liking red line were presented on the anterior surface of the iris and they were not obvious. CONCLUSION: Anterior chamber inside lens coriaceous can induce iris new blood vessels.

Lack of Association between Genetic Polymorphisms Affecting Autonomic Activity and Coronary Artery Spasm

Coronary artery spasm （CAS） is one of the leading pathological causes of a wide spectrum of ischemic heart diseases, ranging from variant angina pectoris to acute myocardial infarction and even sudden cardiac death[1]. Furthermore, Pierron et al. concluded that CAS of angiographically normal or sub-normal arteries is responsible for death or myocardial infarction in 11.6% of all cases. Oddly, the incidence of CAS is remarkably higher in Asians than in Caucasians[3], suggesting genetic involvement In its pathogenesis.

Endoscope-assisted goniosynechialysis combined with phacoemulsification and intraocular lens implantation to manage primary angle-closure glaucoma

AIM: To describe and evaluate a new ophthalmic endoscope surgical technique combined with phacoe-mulsification and intraocular lens (IOL) implantation to treat goniosynechialysis and manage primary angle-closure glaucoma (PACG). ·METHODS: Endoscope-assisted goniosynechialysis combined with phacoemulsification and IOL implantation was performed in 32 eyes of 29 patients with PACG. Regular follow-up was performed 1 week and 1 month, 3, and 6 months after surgery to assess complications, intraocular pressure (IOP), anterior chamber depth, visual acuity, and anterior chamber angle. ·RESULTS: Preoperative mean IOP was 24.88±7.22mmHg with pharmacological treatment, and was 13.70 ±4.02, 13.06 ±3.74, 14.29 ±4.70, and 14.33 ±5.01mmHg 1 week, 1 month, 3 months, and 6 months after surgery, respectively. The postoperative decrease in IOP was significant (P 【 0.05). The rate for all eyes with IOP of 21mmHg or less was 93.8% (30 eyes) at the final visit without ocular hypotensive agents. The average preoperative anterior chamber depth was 1.60 ±0.64mm, and this value significantly increased to 2.72±0.62, 2.76±0.70, 2.73±0.68, and 2.74 ±0.71mm at 1 week, 1 month, 3 months, and 6 months, respectively. Visual acuity was improved in 28 eyes (87.5% ) at 6 months postoperatively. The anterior chamber angle had increased in 25 eyes (78.1% ) at the final visit; it was adhesive 90°-180° in 6 eyes, 180°-270° in only 1 eye. Two eyes exhibited minimal hyphema in the early postoperative period, but it could gradually be absorbed. Fibrinous reaction was observed in five eyes and spontaneously disappeared within 7 days. No shallow anterior chamber, iridodialysis, choroidal deta-chment, or malignant glaucoma was found in any eyes.CONCLUSION:Endoscope-assisted goniosynechialysis combined with phacoemulsification and IOL implantation to manage PACG has several advantages, including optimized visualization, greater accuracy, and improved safety. Our results suggest that it has certain curative effects and clinical application value.

Endoscopy-assisted vitrectomy in the anterior vitreous

Endoscopy-assisted ocular surgery is a relatively old technique that is increasingly being recognized for its application in cases of vitreoretinal disease. This technique is especially useful when both the vitreous and retina are difficult to access because of media opacity, a small pupil, or a microcornea. In this context, the anterior vitreous is often difficult to dissect because of its complex pathological changes. This article reviews the common anatomical features and pathologies that are observed in the anterior vitreous, as well as the applications and indications of endoscopy-assisted vitrectomy in the anterior vitreous.

Ethical issues in penile transplantation

This article provides an overview of the ethical issues associated with penile transplantation, a form of composite tissue allografting. There is only one reported case of human penile transplantation, and, as such, this technique is considered to be experimental. The ethical issues at stake involve both the graft donor and the graft recipient. With regard to the recipient, there are significant concerns relating to surgical risks and benefits, informed consent, body image （including surgical expectations and outcomes） and compliance. Donor issues may include family consent and privacy, as well as graft harvesting （leaving the donor cadaver without a penis）. Many of these ethical issues can be explored during the recipient＇s assessment and consent process. Because no medium-term or long-term outcome data for this procedure exist---only one such operation has ever been performed--the burdens and ethical issues concerning penile transplantation remain unknown.