BACKGROUND:Platelet-rich plasma contains a variety of stimulating factors, and can also raise the proteoglycan and col agen synthesis. OBJECTIVE:To observe the effect of platelet-rich plasma on the proliferation of goat bone marrow mesenchymal stem cel s. METHODS:Blood samples were extracted from the jugular vein of Inner Mongolia Ximeng goats to harvest platelet-rich plasma using centrifugation method. Then, bone marrow was extracted from the goat’s ilium by puncture method to isolate and purify goat bone marrow mesechymal stem cel s using density gradient centrifugation method. After that, primary cel s at good state were cultured in L-DMEM complete medium containing 10%, 20%, 30%platelet-rich plasma or in simple L-DMEM complete medium. RESULTS AND CONCLUSION:Within 2-6 days of culture, cel s in the platelet-rich plasma groups proliferated faster than those in the control group, and with the increasing of platelet-rich plasma concentration, the cel s grew faster, with larger number and more mature morphology. At 4 days of culture, the cel doubling time was about 50, 35, 25 hours in the 10%, 20%, and 30%platelet-rich plasma groups, respectively. These findings indicate that goat platelet-rich plasma can dramatical y promote the proliferation of bone marrow mesenchymal stem cel s in a concentration-dependent manner.

BACKGROUND:Diabetes as a systemic metabolic disease induces the disorders of carbohydrates, fat, and protein metabolism, simultaneously easily causes the lesion of surrounding capilaries, and impacts nutrient metabolism of multiple organs including intervertebral discs. OBJECTIVE:To summarize the research progress of diabetes effects on disc degeneration al over the world. METHODS: The first author used the computer to retrieve the information from PubMed and China National Knowledge Infrastructure. The key words were “intervertebral disc, degeneration, diabetes melitus” in English and Chinese. 8 414 relevant articles were found, which were published from January 1981 to January 2014. Repetitive studies were excluded and 34 articles were in accordance with the inclusion criteria. RESULTS AND CONCLUSION: Pathogenesis of diabetes complications was very complicated. Apoptosis is a hot focus in recent years. Hyperglycemia has been a inducer for apoptosis in intervertebral discs. Diabetes easily leads to systemic smal vessel disease, especialy vascular bud contraction in the terminal plate of vertebral body, and results in regional blood flow decrease or interruption. Thus, the nutrient substance carrying along the end plate was reduced, which resulted in disc dystrophy and degeneration. The reduction in extracelular matrix in the intervertebral discs is a major reason for disc degeneration. The mechanisms underlying diabetes effects on disc degeneration remain unclear and deserve further investigations.

Objective To evaluate pressure-flow study in assessment of dysuresia symptom in elderly male patients. Methods A total of 125 elderly male patients with dysuresia and prostatic enlargement underwent urodynamic examination.Their age ranged from 56 to 83 years with a mean of 68 years.If the patients urinated smoothly with satisfying urinary flow curve,the pressure-flow study would be performed on them.According to the results,the patients were classified as 3 groups:BOO,equivocal BOO and non-BOO groups.Their detrusor contractive function was classified as strong,normal,weak or very weak. Results Of the 125 patients undergoing urodynamic test,87 obtained definite pressure-flow study findings.Of the 87 patients,39 were with BOO,18 with equivocal BOO and 30 without BOO.In the BOO,equivocal BOO and non-BOO groups,detrusor pressure at maximum flow was (99.2?34.3) cmH 2O (1cmH 2O=0.098 kPa),(46.9?9.9)cmH 2O and (30.8?10.0)cmH 2O,respectively;intravesical opening pressure was (99.4?39.6) cmH 2O, (43.7?9.9) cmH 2O and (29.9?9.7) cmH 2O, respectively; minimum voiding detrusor pressure was (61.3?27.5) cmH 2O, (33.9?14.1)cmH 2O and (22.1?12.5)cmH 2O,respectively;and maximum detrusor pressure was (113.0?42.1)cmH 2O,(55.8?14.9)cmH 2O and (38.4?11.3)cmH 2O,respectively.The detrusor function was normal or strong in 74.4% (29/39) of patients with BOO,27.8%(5/18) of patients with equivocal BOO and 26.7% (8/30) of patients without BOO.All these parameters of the BOO group were higher than those of the equivocal BOO group and non-BOO group (both P

Objective To evaluate the bladder functions by urodynamic studies in diabetic patients with lower urinary tract symptoms (LUTS). Methods Urodynamic studies were performed in 42 diabetic patients (24 men and 18 women;age range,38-78 years).The diabetic history of the patients was from 1 month to 25 years. Results Of the 41 cases who underwent all items of urodynamic examination,38 cases (93%) had abnormal findings and 3 (7%),normal.The detrusor was underactive in 14 cases (34%) and areflexia in 10 (24%);BOO was found in 13 (32%).One female patient had stress urinary incontinence. Conclusions There is a high prevalence of bladder dysfunction in diabetic patients with LUTS.It is necessary to perform urodynamic studies in diabetic patients before initiation of therapy,especially in patients who are assigned to undergo bladder and urethral surgery.Preoperative urodynamic studies can contribute to the surgical success rate.

BACKGROUND:Intervertebral disc can bear load but lack vessels. Nucleus pulposus cel s have the problem of phenotype loss during in vitro culture that can lead to degenerative changes. The mechanism of intervertebral disc degeneration remains unclear.
OBJECTIVE:To explore the approaches of isolation, adherence culture, amplification and identification of the rabbit nucleus pulposus cel s in vitro, to observe the growth characteristics of nucleus pulposus cel s in different generations.
METHODS:Type Ⅱ col agenase digestion method plus explants culture method was used to isolate and purify nucleus pulposus cel s and then amplify in vitro. The morphology and growth of primary and passaged cel s was observed under the inverted microscope, the number of cel s was counted and the growth curve was draw. The morphology of the cel s was observed under light microscope after hematoxylin-eosin staining, and the expressions of col agen type Ⅱ and aggrecan were examined by immunocytochemistry.
RESULTS AND CONCLUSION:Nucleus pulposus cel s of rabbit were isolated, cultured and amplified in vitro successful y. Growth activity was observed, and found that the 1-3 generation nucleus pulposus cel s proliferated more rapidly and vigorously. The proliferation of nucleus pulposus cel s was decreased while the cel passaged more generations. These isolated and cultured nucleus pulposus cel s could positively express the col agen type Ⅱ and aggrecan. In vitro combination of type Ⅱ col agenase digestion method and explants culture method could obtain high purity nucleus pulposus cel s, and the cultured nucleus pulposus cel s were grew in round or polygonal. The 1-3 generation of cel s had strong activity.

BACKGROUND:Percutaneous kyphoplasty for osteoporotic vertebral fractures with posterior vertebral wal breakage can relieve pain rapidly, but there is a risk of leakage of bone cement. OBJECTIVE: To observe the effect of bone filing mesh container in percutaneous kyphoplasty for osteoporotic vertebral fractures with posterior vertebral wal breakage. METHODS: Forty senile patients with osteoporotic vertebral compression fractures were enroled, whose CT and MRI showed different degrees of posterior vertebral wal breakage, including 19 males (21 vertebral bodies) and 21 females (28 vertebral bodies), aged 50-87 years. These 40 patients were subjected to percutaneous kyphoplasty, and bone filing mesh container was used to deliver bone cement. Then, changes in visual analogue scale score, vertebral height and leakage of bone cement were observed in patients before and after treatment. RESULTS AND CONCLUSION:The surgery was successful in al the 40 patients, and no pulmonary embolism, cement leakage, and spinal cord and nerve root injuries appeared. Al the patients were folowed for 10-12 months. The visual analogue scale scores and vertebral height were improved significantly at both 1 week and 3 months after treatment compared with those before treatment, but there was no difference in the visual analogue scale scores and vertebral height at 1 week and 3 months after treatment. These findings indicate that percutaneous kyphoplasty with bone filing mesh container is effective to prevent bone cement leakage.

Objective To understand the role of increasing peripheral blood platelet count in the diagnosis and prognosis of patients with active pulmonary tuberculosis.Methods This analysis included 80 inpatients with active pulmonary tuberculosis evidenced by positive sputum smear,43 patients in recovery and 89 healthy controls during the period from January to June in 2012.Peripheral white blood cell,platelet count,and neutrophil percentage were assayed.Erythrocyte sedimentation rate (ESR)and serum C-reactive protein (CRP)were measured and compared.The data were analyzed by using SPSS 13.0 software.Results ESR and CRP values were significantly higher in the patients with active pulmonary tuberculosis than in the patients in recovery (P=0.006 5 and P=0.007 3,respectively).The peripheral blood platelet count exceeded normal range in 41.94% of the patients with active pulmonary tuberculosis,which was significantly higher than that in the patients in recovery (P=0.001 4)and controls (P=0.000 0).Platelet count was positively correlated with CPR (r=0.515,P<0.000 1)and ESR (r=0.398,P<0.001)value.However,peripheral white cell count and neutrophil count were not different from the corresponding values of controls.Conclusions Increasing peripheral platelet count may play a role in assessment of tuberculosis disease activity and the effect of anti-tuberculosis treatment.

Objective To understand the bacteriology and the association between drug susceptibility and clinical features from recent hospitalization of tuberculosis(TB)patients with positive tuberculosis bacilli culture.Methods We collected the clinical data of inpatients due to tuberculosis or pulmonary disease with positive tuberculosis bacilli by BACTEC960 culture auto-analysis system and possessing anti-tuberculosis drug susceptibility testing results from January 2008 to November 2008.All isolated strains were tested with first-line drugs included Streptomycin(S),Isoniazid(H),Rifampicin(R)and Ethambutol(E).Some isolate strains were used to test Amikacin(Am),Capreomycin(Cm)and Ofloxacin(Ofx).We recorded the drug-tested results and clinical data and retrospectively analyzed them.The patients with pulmonary disease from nontuberculosis mycobacteria(NTM)were excluded.Results (1)There were 417 patients with positive culture of tuberculosis bacilli (included 294 male and 123 female).The mean age was(47.8 18.1)years(ranging from 6 to 91 years).There were 68 cases complicated with endobronchial tuberculosis(EBTB)and 56 cases with type 2 diabetes mellitus.(2)There were 271 cases for initial treatment and 146 cases of relapsing tuberculosis.The total drug resistant rate was 53.5 percent,and of oflx was as high as 31.86 percent.The initial drug resistant rate was S 22.5%,H 25.8%,R 17.3%,E 21.0%,Am 6.3%,Cm 10.0%,Ofx 16.6% respectively and the required rate was 67.8%,82.9%,68.5%,68.5%,19.9%,25.3%,58.2% respectively.(3)There were 143 patients with multi-drug resistant TB(MDR-TB).The mean age 44.59?16.31 was significantly younger than of other patients(P

AIM: Site-specific functional neurons of brains were with different cellular morphology. It has not been fully understood whether the grafted neural stem cells could differentiate into the site-specific neurons. This experiment is to investigate the neuronal differentiation of the neural stem cells derived from a human fetal brain after transplanted into young rats' brains, to study the possibility of cell-replacement therapy for children's brain disorders with neural stem cells. METHODS: Experiments were performed at the Cell Laboratory of Naval General Hospital from April to July 2007. ①Human fetal brain tissues of 16 week gestation were provided by Department of Gynaecology and Obstetrics of Naval Hospital. Pregnant woman and family members signed an informed consent. Experimental intervention was approved by Hospital Ethical Committee. Fourteen clean brood young SD rats aged 10 days, irrespective of gender, were provided by Experimental Animal Center of Medical College of Peking University. Animal intervention met the animal ethical standards. ②The neural stem cell spheres were derived from the fetal brain tissues of 16 week gestation. The differentiation multipotency of the neurosphere was identified when cultured in a child's cerebrospinal fluid (CSF). The neurospheres cultured in vitro for 14 days were injected into the lateral ventricles of young rats of 10 days old. The rats were respectively killed at days 4, 7 and 14 after transplantation. The special immuno-fluorescent assays were performed using anti-human neurofilament (anti-hNF) to show the location and morphology of graft neurons. RESULTS: ①The typical floating neurospheres were obtained, with the potency to differentiate into neurons, astrocytes and oligodendrocytes. ②The neuronal differentiation of grafts was detected with the mixture of three monoclonal antibodies against human neurofilament. Four days after transplantation, the immune response positive cells lied within the granule cell layer of cerebral cortex were shown in the shape of granule cells, or within the pyramid cell layer in the shape of pyramid cells with long processes, and the interneuron-like cells also were seen. The Purkinje cells arranging in a monolayer were detected in the cerebellum. Compared the results at different time points, the location of grafts were the same. The graft cells were less and the processes were longer over time. CONCLUSION: The in vitro cultured neurosphere cells can migrate into brain tissues and differentiate into site-specific neurons in shape after transplanting into the lateral ventricles of young rats. It is suggested that the host brain tissue microenvironment played an important role in guiding the graft differentiation into neurons. The results have an important significance for understanding cell replacement of developing brain disorders.

BACKGROUND:Transplantation of mesenchymal stem cels to prevent and treat degeneration of the intervertebral disc is a feasible method. Mesenchymal stem cels co-transfected by SRY-related high mobility group-box gene 9 (SOX-9) and growth differentiation factor-5 (GDF-5) can differentiate into nucleus pulposus cels, in order to obtain greater effect of induction and proliferation of nucleus pulposus cels. OBJECTIVE:To investigate the effect of SOX-9 and GDF-5 co-transfection on the differentiation of rabbit bone marrow mesenchymal stem cels into nucleus pulposus cels. METHODS: We separated and cultured bone marrow mesenchymal stem cels from the bone marrow of rabbit aged 4 months. Passage 3 cels were divided into five groups andin vitro induced to differentiate into nucleus pulposus cels: non-transfected group, empty vector transfection group, SOX-9 transfection group, GDF-5 transfection group, SOX-9 and GDF-5 co-transfection group. At 14 days after transfection, RT-PCR was employed to assay SOX-9, GDF-5 and colagen type II mRNA expressions in bone marrow mesenchymal stem cels. The marker of nucleus pulposus cels-KRT19 expression was also detected by immunohistochemical staining. RESULTS AND CONCLUSION:In the co-transfection group, the mRNA expressions of SOX-9, GDF-5, and colagen type II were significantly higher than those in the SOX-9 transfection group, GDF-5 transfection group, and both these two groups, respectively (P < 0.05). Cels were positive for KRT19 in the SOX-9 and GDF-5 groups, and strongly positive for KRT19 in the co-transfection group. These findings indicate that double gene-transfected bone marrow mesenchymal stem cels are better than single gene-transfected cels with regard to differentiation into nucleus pulposus cels and secretion of extracelular matrix.