Objective To examine the effects and mechanisms of siRNA targeting aquaporin 4 (AQP 4) in combination with hyperbaric oxygen therapy(HBO) on cerebral edema and apoptosis in the brain tissue of rats after hemorrhage.Methods Rats were randomly divided into four groups,the control group,the hyperbaric oxygen group,the AQP-4 siRNA group and the combination therapy group (24 rats).Thrombin Ⅶ was injected into the caudate nucleus to establish the hemorrhage model.Construction of siRNA targeting aquaporin 4 was conducted.The mRNA expression of AQP-4 was detected by RT-PCR at day 3.Changes in brain moisture and blood-brain barrier perme ability were measured by a wet/dry weight method and Evans blue fluorometry.The nerve cell apoptosis rate was analyzed by Annexin V andTdT-mediated dUTP-biotin nick end labeling (TUNEL).The expression of proteins including AQP-4,MMP-2,MMP-9,Bcl-2 and caspase-3 was detected by Western Blotting.All the animals were given a score for their nerve function at day 3.Results AQP-4 siRNA treatment obtained better effects than HBO in decreasing the brain edema leveland silencing AQP-4 mRNA(P＜0.05)while,the combination therapy group achieved the best results(P＜ 0.05).Compared with the control group,the percentage of apoptotic cells decreased in all the three treatment groups,with the most marked decrease observed in the combination treatment group(4.24± 0.04)％(F=13.76,P=0.001).The expression of AQP-4,MMP-2,MMP-9 and caspase-3 was lower (P＜0.05) and the expression of Bcl-2 was higher(P＜0.01)in the combination treatment group than in the other three groups.Compared with the control group,all the other three groups received better scores on nerve function defect evaluation at day 3 after hemorrhage(P＜0.05),with the combination treatment group again achieving the most favorable score (4.7 ± 1.1) (F=7.21,P =0.013).Conclusions Targeted siRNA interference combined with hyperbaric oxygen can effectively reduce cerebral edema after cerebral hemorrhage,inhibit neuronal apoptosis and promote neuron function recovery.The underlying mechanisms may be related to down-regulation of AQP-4,MMP 2,MMP-9 and caspase-3 expression and up-regulation of Bcl-2 expression.

Objective To evaluate the technique and outcome of arthroscopic reconstruction of posterior cruciate ligament(PCL)with preservation of the remnants.Methods From January 2006 to January 2009.16 knees in 16 patients with PCL tear were verified with arthroscopy.All the damaged PCL were reconstructed with single bundle of autogenous quadrupled hamstring tendons under arthroscopy without remove of the remnants of PCL.Endo-button and Bio-absorbabh interference screw were used for direct anatomic fixation of the reconstructed ligament.Results No severe complication occurred in early stage after operation in the 16 patients.All of them were followed up from 4 to 14 months with an average of (7.4±2.1)months.Lysholm score was remarkable improved from mean(61.3±8.7)preoperative scores to mean(91.7±6.3)scores at the latest follow up(P＜0.05).Furthermore,there was a significant improvement in IKDC scofe from abnormal (grade C)in 9 knees and severely abnormal (grade D)in 7 knees preoperatively to normal(grade A)in 11 knees,nearly normal(grade B)in 5 knees at the follow up.All of the 16 patients returned to the same sports level before injury.Conclusion Arthroscopic reconstruction of PCL with preservadon of the remnants has advantages of minimal inmmrticular interference,and potential facility to revagculization and reinnervation of the reconstlucted ligament.

Activating blood circulation to remove stasis method is an important therapy of TCM, which can be matched with various methods, as qi-supplementing, qi-regulating, heat-clearing with detoxication, meridian warming, wind-dispelling to remove dampness, yin-nourishing, phlegm-dissolving to alleviate depression and visceral dredging by purgation, to produce various effects on angiogenesis. Its positive or negative impacts on quality or quantity of angio-genetic regulatory factor play a crucial part for the effects. This article explores the effect and mechanism of activating blood circulation to remove stasis method on angiogenesis bi-directionally.
Angiogenic Proteins ; metabolism ; Blood Circulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hematologic Agents ; pharmacology ; therapeutic use ; Hematologic Diseases ; drug therapy ; Humans ; Medicine, Chinese Traditional ; Qi

Background and purpose:Carcinoma of the kidney is the most common malignant renal parenchymal carcinoma;its biological behavior is extremely complicated and is not sensitive to radiotherapy and chemotherapy.Studies have shown that the hypoxia-inducible factor HIF1-? and HIF2-? are related to the occurrence and development process of the clear-cell carcinoma of kidney.So,we intend to construct psilencer3.0-HIF-? siRNA recombinant plasmid in this study through RNA interference method,thereby providing an effective tool for further exploration of the role of HIF in the occurrence and development of clear-cell carcinoma of kidney.Methods:Design and chemically synthesize the DNA fragments of coding HIF-1? and HIF-2? siRNA and make them up into siRNA expression vector through gene recombination.Adopting the real-time quantitative PCR and Western blot tested the inhibitive effect of the constructed siRNA expression vector on mRNA and protein levels on the target gene expression.Results:After 786-0 cells transfecting with psilencer3.0-HIF-1?siRNA and psilencer3.0-HIF-2?,the inhibitory rate of mRNA expression of HIF-1? and HIF-2? reached 82.1% and 87.4% respectively,and OS-RC-2 cells transfecting psilencer3.0-HIF-1? siRNA and psilencer3.0-HIF-2?,the inhibition rate reached 91.2% and 81.2% respectively.The protein expression of the experimental group with 786-0 cells and OS-RC-2 cells transfecting HIF-1? interference plasmid and HIF-2? interference plasmid were lower at different levels than that from the blank control group.Conclusions:The constructed siRNA expression vector can effectively inhibit the expression of target gene HIF-1 and HIF-2 at mRNA and protein levels.

Low back pain is becoming an important factor affecting people's quality of life, while the age of its onset is getting younger and younger, and the social and economic losses caused by low back pain are huge every year. Intervertebral disc degeneration (IDD) is an important cause of low back pain. Due to multiple factors, biomechanical and structural changes occur in intervertebral disc tissue, including rupture of annulus fibrosus, protrusion of nucleus pulposus, which cause compression of spinal cord and nerve root, and lower back pain. Micro-RNA (miRNA) is a kind of single-stranded non-coding small molecule RNA, with 18-24 nucleotides in length, which exists widely in eukaryotes. As one of the important regulatory molecules of gene expression, it has been proved to play a key role in the initiation and progression of many diseases, and it may also play an important role in intervertebral disc degeneration. At present, the clinical treatment for IDD is mainly surgical treatment to alleviate clinical symptoms. Even if surgical treatment can achieve good results, it will bring great physical trauma and economic burden to patients. The role of miRNA in IDD is one of the hotspots in the current academic research. Studies have shown that miRNA has abnormal expression patterns in degenerative intervertebral disc tissues and participates in a variety of pathological processes of IDD. At present, some miRNAs have been proved to be related to a variety of pathological processes in IDD, including nucleus pulposus cell apoptosis and proliferation, extracellular matrix degradation, autophagy, inflammation and cartilage endplate degeneration. The comparative study of gene chip showed that there were significant differences in the expression of some miRNAs between degenerative and normal nucleus pulposus cells. These differentially expressed miRNAs may be involved in the process of nucleus pulposus cell degeneration by regulating their respective upstream or downstream pathways. Most of the regulatory pathways are crossed and parallel, thus constructing a huge miRNA regulatory network. Understanding the target genes and mechanisms of miRNA in the pathogenic process can provide an important reference for the origin, development and prognosis of IDD. In this article, the important role of miRNA in IDD and the potential significance of clinical treatment are reviewed. With the in-depth study of miRNA and the molecular biological mechanism can provide new ideas for the diagnosis and treatment of IDD, which is likely to become a new strategy for biological treatment of IDD.

Objective To test the safety and effectiveness of transpedicular fixation combined with transpedicular bone grafting via less invasive paraspinal intermusclar approach in treatment of thoracolumbar fractures.Methods The study involved 23 cases of thoracolumbar fractures treated with paraspinal multifidus intramusclular mini-incision,transpedicular bone grafting,and short-segment pedicle screw fixation from June 2009 to June 2012.There were 16 males and 7 females at age of 19-55 years (average 38.8years).Time from injury to surgery varied from 6 hours to 7 days (average 3.2 days).Fracture level was T11 in three cases,T12 in seven,L1 in nine,and L2 in four.According to Denis fracture classification,there were altogether 10 compression fractures and 13 burst fractures.McCormack load sharing classification scored average 5.3.Before operation,anterior vertebral body height ratio was average 58.6％(range,45 ％-73％) and kyphosis angle was average 23.7° (range,15°-34°).Results Operation lasted for average 95.5 minutes (range,75-130 minutes) with intraoperative bleeding of average 160.3 ml (range,115-220 ml).Unilateral incision that was averaged 3.5 cm (range 3.2-4.0 cm) in length obtained primary healing.Average follow-up time was 12.6 months (range,7.5-18 months).Average height of the anterior border was corrected to 97.3％ and average kyphosis angle was corrected to 4.6°.There was neither instrumentation failure nor symptom of persistent postoperative back pain.Conclusions Transpedicular fixation with transpedicular bone grafting via paraspinal muscle approach provides effective recovery of vertebral morphology and correction of kyphotic deformity.Furthermore,the technique gains advantages of easy operation,small trauma,less blood loss and rapid recovery.

Objective To explore the clinical manifestations of ultrasonography(US),99mTc-methoxyisobutylisonitrile(99mTc-MIBI),and 131I whole body scan(131I-WBS)of patients with differentiated thyroid carcinoma(DTC)and cervical lymph node metastasis.Methods A total of 117 cases received examinations of US and 99mTc-MIBI before uses of 131I and received 131I-WBS after 6 d of 131I treatment.Combined with serum thyroglobulin(Tg),131I-SPECT/CT,lymph node biopsy and clinical follow-up of patients,cervical lymph node metastases were identified.Results In 117 patients,the sensitivity of US,99mTc-MIBI and 131I-WBS were 76.5%,67.1% and 83.5%(P

OBJECTIVETo evaluate the clinical effectiveness of transurethral seminal vesiculoscopy (TUSV) combined with finasteride in the treatment of recurrent hemospermia.

METHODSThis study included 32 patients with recurrent hematospermia, with the disease course of 3 months to 4 years. After administration of finasteride at 5 mg/d for 2 weeks, the patients underwent TUSV for both exploration of the causes and treatment, followed by medication with finasteride at the same dose for another 2 weeks. Postoperative follow-up was conducted for observation of the outcomes and complications.

RESULTSTUSV was successfully accomplished in all the 32 cases, which revealed 16 cases of seminal vesiculitis, 10 seminal calculi, 1 seminal vesicle cyst, 2 seminal vesicle polyps, and 3 seminal vesicle abscess. The operative time was 20 to 51 (31.0 +/- 5.2) minutes. Postoperative complications included 1 case of acute epididymitis and 3 cases of breast discomfort within the first 4 weeks. No incontinence, urethral stricture, rectal injury, retrograde ejaculation, and sexual dysfunction occurred postoperatively. All the patients but 1 were followed up for 6 months to 2 years. Twenty-nine of the cases were cured, and 2 experienced recurrence.

CONCLUSIONTransurethral seminal vesiculoscopy combined with finasteride is safe and effective for the treatment of recurrent hemospermia.

Adult ; Endoscopy ; methods ; Finasteride ; therapeutic use ; Follow-Up Studies ; Hemospermia ; therapy ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Objective To investigate the treatment effects of operation and nerve growth factor on adult multisegmental cervical spinal cord injury without fracture dislocation.Methods Sixty-eight patients with multiple segmental cervical spinal cord injury without fracture dislocation,admitted to our hospital from January 2004 to May 2011,were chosen in our study; according to the will of the patients,18 patients received conservative treatment (group A),25 patients were treated by posterior single open-door laminoplasty (group B) and the other 25 patients were treated with posterior single open-door laminoplasty combined with nerve growth factor (group C,once daily for 4 weeks); their clinical data were retrospectively analyzed.Follow up was performed at 3,6 and 12 months after the treatments;Japanese Orthopaedic Association (JOA) scale was employed to evaluate the clinical efficacy.Results Three,6,12 months after the treatments,the JOA scale scores were statistically different among the three groups (P＜0.05); the scores in group B and C were significantly higher than those in group A at all time points (P＜0.05); 6 and 12 months after the treatmemts,the JOA scale scores in group C were significantly higher than those in group B (P＜0.05).Conclusion To patients with multisegmental cervical spinal cord injury without fracture dislocation,conservative treatment can make the spinal cord function partially restored,but the effect is limited; exogenous nerve growth factor has good repairing effect on spinal cord injury.

Objective:To investigate the effect and mechanism of bone marrow mesenchymal stem cell (BMSC)-derived exosomal microRNA-1297 (miR-1297) on hippocampal neuron damage in depressed rats.Methods:BMSCs and BMSCs-derived exosomes were prepared and identified. Rats were first injected with corticosterone to establish the model of depression, and then injected with BMSCs-derived exosomes. Superoxide dismutase (SOD), malondialdehyde (MDA), lactate dehydrogenase (LDH), TNF-α and IL-1β in rat serum samples, hippocampal tissues and neurons were detected. Expression of miR-1297 in hippocampal tissues and neurons was detected by RT-qPCR. A rat hippocampal neuron injury model was established to investigate the role of BMSC-derived exosomes and miR-1297 in neuronal apoptosis and proliferation. The targeting relationship between miR-1297 and connective tissue growth factor (CTGF) was analyzed using dual luciferase reporter genes.Results:In the hippocampus of depressed rats, the expression of miR-1297 was low, while the expression of CTGF was elevated. Exosomes derived from BMSCs can inhibit the expression of CTGF by up-regulating the level of miR-1297, thereby inhibiting neuronal cell apoptosis in the hippocampus of depressed rats, while increasing the level of SOD, and reducing inflammatory damage, and ultimately improving the behavioral function of depressed rats.Conclusions:Depressed rats showed decreased expression of miR-1297 and increased expression of CTGF. BMSC-derived exosomes inhibited CTGF expression through up-regulating miR-1297, thereby improving hippocampal neuron damage in rats with depression.