Objective To investigate the effect on the differentiation of bone marrow mesenchymal stem cells (BMSC) with non-contact co-culture with mechanical stimulated ligament fibroblasts. Methods A cyclic 10% uniaxia strain at 1 Hz was applied on rat pelvic ligament fibroblasts, then were co-cultured with BMSC for 3, 6 and 12 days in non-contact condition. The protein expression of collagen Ⅰ ,Ⅲ in BMSC were detected by SP method and revealed by the mean gray value. The mRNA expressions of collagens type Ⅰ and type Ⅲ in the BMSCs were measured with real-time (RT)-PCR ,and the results were indicated by the ratio between the mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) . Results (1) Protein expression; after 3 days co-culture with pelvic ligament fibroblasts, expression of collagen Ⅰ and Ⅲ in BMSC are 82. 4 ± 3. 4 and 76. 8 ± 2. 5. When compared with 80. 2 ± 2. 6 and 74. 6 ± 1. 1 in BMSC without co-culture, there was no significant difference (P > 0. 05) . After 6 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 126. 6 ±2. 2 and 118. 6 ± 1. 4 in BMSC were significantly higher than 82. 7 ±3. 0 and 76. 2 ± 1. 3 in BMSC without co-culture (P < 0. 05). Similarly, after 12 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 135. 3 ±3. 4 and 128. 7 ± 2. 6 in BMSC were significantly higher than 86. 6 ± 1. 3 and 81. 8 ± 1.4 in BMSC without co-culture (P <0.05). (2)mRNA expression:after 3 days co-culture with pelvic ligament fibroblasts , the mRNA expression of type Ⅰ and type Ⅲ collagens in BMSC are 2. 10 ±0. 20 and 1. 20 ±0. 30. When compared with mRNA expression of 2. 01 ±0. 12 and 1. 13 ±0.21 in BMSC without co-culture, no significant difference were observed (P > 0. 05). After 6 days co-culture with pelvic ligament fibroblasts , the mRNA expressions of type Ⅰ and type Ⅲ collagens mRNA were 5. 60 ±0. 21 and 2. 61 ±0. 20, which were significantly higher than 3. 70 ±0. 33 and 1. 82 ± 0. 14 in BMSC without coculture (P < 0. 05). After 12 days co-culture with pelvic ligament fibroblasts, the mRNA expressions of type Ⅰ and type Ⅲ collagens of 5. 91 ±0.31 and 2. 92 ±0. 23 were significantly higher than 4. 04 ±0. 21 and 2. 04 ±0. 13 in BMSC without co-culture (P <0. 05). Conclusion Non-contact co-culture with mechanical stretch stimulated ligament fibroblasts, it might promote synthesis of types Ⅰ and Ⅲ collagen in rat BMSCs and induced BMSC differentiated into pelvic ligament fibroblasts.

Objective To explore the hemorheological changes in children with essential hyepertension,and explore their clinical significance.Methods The hemorheological parameters,blood lipid,blood glucose in 37 children with essential hyepertension [19 boys and 18 girls,median age(12.49?3.20) years] were measured and analyzed.The parameters in 30 healthy children were measured as controls.SPSS 11.5 software was used to analyze the data.Results The whole blood viscosity under high,middle and low shear rate,plasma viscosity,hematocrit,rigidity index of erythrocyte,platelet aggregation M were significantly higher in hypertension group than those in healthy control group(Pa0.05).Conclusions There have been developed significant hemorheological changs in children with essential hypertension.The hemorheological changes might play an important role in pathogenesis of childhood hypertension.

Background Vitretomy and lenstomy with silicone oil tamponade is an effective method for complicated vitreous retinopathy.The severe anisometropia after silicone retention is usually treated by two-point transscleral suture fixation for posterior chamber intraocular lens(IOL)implantation.In order to reduce the number and difficulty and complication of the operation,the surgical method should be improved.Objective The goal of this study was to observe the resuh of silicone oil removal combined with four-point trans-scleral suture fixation of posterior chamber IOL after vitrectomy.Methods A retrospective case-observational study design was adopted.Twenty eyes with silicone oil tamponade from 20 patients without lens and posterior capsule after vitrectomy were included in this study.Silicone oil removal with four-point trans-scleral suture fixation of posterior chamber IOL was performed.The anterior ocular inflammatory response,intraocular pressure,uncorrected and corrected acuities before and after operation,corneal endothelial cell counting and postoperative complications were observed and analyzed.Written informed consent was obtained from all patients prior to the operation.Results All of the operative eyes in this study showed improvement of visual acuity after operation.Of the 20 eyes,a visual acuity of ≥0.8 was seen in 2 eyes,0.6-0.7 in 6 eyes,0.3-0.5 in 8 eyes and 0.05-0.2 in 4 eyes 3 months after the removal of silicone oil.The uncorrected acuity postoperation was significantly improved in comparison with preoperation(H=10.147,P＜0.01),but no significant difference was found in the corrected acuity between preoperation and postoperation(X =2.089,P＜ 0.01).The number of the corneal endothelial cells was(2064±329)cells/mm2 before operation,and that after operation was(1987±269)cells/mm2,showing an insignificant change between them(t =1.660,P ＞ 0.05).No abnormality of IOL position was found in all 20 operated eyes.There was not serious postoperative complication in all 20 patients.Conclusions The combination of silicon oil extraction with four-point transscleral suture fixation IOL is effective in eyes without posterior capsule or lens after vitrectomy.It can reduce the operation time and improve the postoperative acuity and the quality of life of patients.

Objective To explore the effects of repetitive transcranial magnetic stimulation on behaviors and hippocampus neuronal apoptosis in rats with chronic stress-induced depression.Methods 40 male SD rats of SPF grade were randomly divided into normol control group (n =8) and model preparation group (n =30) after screening.Rats in model preparation group were singly housed and given chronic unpredictable mild stress(CUMS) to build depression model.Excluding unsuccessful modeling rats,the model preparation group was divided into three groups:model group(n=8,without any treatment),rTMS group (n=8,with the intervention of 10 Hz rTMS) and shame group (n=8,simulation of rTMS environment without rTMS stimulus).The changes of behaviors in each group were detected by weight measurement,sucrose consumption test and open-field test.The changes of morphology of hippocampal neurons were detected by Nissl's staining.The changes of Bax in hippocampal neuron were detected by Immunohistochemical staining.Results (1) Behavioral results showed stress for 21 d could make rat behavior scores decrease significantly(all P＜0.05),and rTMS intervention could significantly improve their behavior scores (all P＜0.01).Compared with model group,the weight reduction rate (0.32±0.05)％,the score of sucrose consumption test(7.03 ± 1.02) and the score of open field test(8212.41 ± 1416.15,8.75 ± 1.58) in rTMS group was higher(P ＜0.01).(2) Nissl staining showed stress for 21 d could make the number of hippocampal CA3 neurons was reduced,cell morphology was poor,and the number of Nissl bodies was reduced.rTMS intervention could increase the number of hippocampal CA3 neurons,cell morphology was integral and the number of Nissl bodies was increased.(3) Immunohistochemistry results showed stress for 21 d could cause the number of Bax cell were significantly increased(P＜0.01),and rTMS intervention can make the number of Bax cell were significantly lower(P＜0.01).Conclusion rTMS intervention improves the depressive behavior in chronic stress depression model rats and inhibits the apoptosis,which might work through inhibition of neuron apoptosis and decline of Bax expression in hippoeampal neurons.

Objective To summarize the experience on diagnosis and treatment of splenic neoplasm. Methods Clinical data of 40 patients with primary splenic space occupying lesion treated in our hospital were retrospectively analyzed. Results Thirty four out of the 40 cases were diagnosed as primary splenic neoplasm preoperatively. The detectable rate of B-US was 94%, and that of CT was 96%. The discrimination rate of benignancy and malignancy by CT was 84%. Serum AKP and ?-GT were significanfiy increased in most of the malignancies. Seventeen among 22 cases with benignancy were treated by splenectomy, and the others underwent partial splenectomy or tumor resection. Sixteen of 18 patients with malignancy underwent splenectomy, and two did biopsy. Pathology revealed cysts in 13, angiocavernoma in 4, inflammatory pseudotumor in 3, caverous lymphangioma in 1, and cystic degeneration of liomyoma in 1; malignant lymphoma and lymphsarcoma in 9, hemangiosarcoma in 3, and fibrosarcoma, liomyosarcoma, malignant fibrous histiocytoma in 1 each. Three cases with malignancy have survived more than 5 years. Conclusions Splenic neoplasm is diagnosed mainly according to clinical manifestation and image examination. The discrimination of benignancy and malignancy depends on CT, angiography, and serous AKP and ?-GT level. Radical operation and complex treatment could improve the prognosis of splenic malignant tumor.

This study was aimed to analyze the environment and situation of patent strategy implementation of manufacturing industry of Chinese patent medicines in G uangdong province . This article pointed out that G uang-dong Chinese patent medicines manufacturing implementation patent strategy faced huge opportunities and chal-lenges. It was suggested that the fully understanding, mastering and using of beneficial conditions as well as con-trolling and eliminating the adverse factors will contribute to the better implementation of Chinese patent medicines manufacturing patent strategy in G uangdong province .

Objective To investigate the evaluation of high-resolution computed tomography (HRCT) in patients with chronic obstructive pulmonary diseases (COPD), and the relationship with the pulmonary function test and BODE index.Methods 32 COPD patients and 22 normal controls were examined by pulmonary function test (PFT) and HRCT. The data of two groups were compared.Results The volume and pixel index (PI) in forced expiration were higher in the COPD group than those in the control group ( P<0.01), but there was no significant difference in Vin between two groups ( P>0.05). In the COPD group, Vex and Vin were negatively correlated with FEV1/FVC ( P<0.05) respectively. There were no correlation between Vin, Vex and FEV1% pre in the COPD patients ( P>0.05) respectively. Vin and Vex were not correlated with BODE index (all P>0.05) respectively. The ratio of Vex/Vin was positively correlated with BODE index ( P<0.05), and not correlated with FEV1/FVC ( P>0.05). PI910ex was positively correlated with BODE index ( P<0.01), and negatively correlated with FEV1%pre, FEV1/FVC (all P<0.05) respectively.Conclusion The pulmonary function of COPD patients can be evaluated by HRCT, especially by PI910ex.

Objective To evaluate the cognitive function of chronic obstructive pulmonary disease (COPD) stable patients with ExtendedScale for Dementia (ESD). Methods 37 patients with stable COPD (COPD group) and 40 healthy persons (control group) were enrolled.The COPD patients were divided into different groups as their PaO2. They were assessed with ESD and Mini Mental Status Examination(MMSE). Results The total score of ESD and the subtest scores of ESD in leaning, memory, calculation, constructive function were significantlylower in the COPD group than in the control group (P<0.001). The total score of MMSE and the subtest scores of MMSE in memory,attention and calculation, short- and long-term memory were significantly lower in the COPD group than in the control group (P<0.05). Thetotal score and the subtest scores in learning, memory, calculation of ESD were lower in patients with PaO2<60 mmHg than those withPaO2≥60 mmHg (P<0.05), same as the total score and the subtest scores in memory, attention and calculation, short- and long-term memoryof MMSE (P<0.05). Both ESD and MMSE correlated significantly with PaO2 (P<0.01). Conclusion ESD can be used as an evaluationtool for cognitive function in stable patients with chronic obstructive pulmonary disease.

Objective To develop a three-dimensional porous film of human hair keratin (HHK)-collagen sponge complex for use as a dermal substitute. Methods The three components F, B, and Z derived from healthy human hair were weaved into a meshwork and integrated with purified soluble type Ⅰ collagen extracted from bovine tendons to prepare a highly porous film with vacuum freeze-drying followed by secondary cross-linking with glutaraldehyde. The film was grafted beneath the dorsal skin in 21 SD rats (experimental group), with simple collagen sponge serving as the negative control. The rats receiving surgical operation but without graft served as the blank control. The graft and its surrounding tissue were harvested on days 3, 7 and at weeks 2, 4, 6, 8, 12 after implantation for evaluation of tissue compatibility, vascularization and degradation. Results The prepared collagen sponge film was semitransparent and porous. Three to 7 days after grafting, inflammatory reaction was relieved gradually, and several fibroblasts and blood vessels were found adherent to the gratts in the experimental groups. At week 4, the wounds healed in the experimental groups, and the fibroblasts were actively secreting collagen and the film degraded obviously with the appearance of elastic fibers. At weeks 6 and 8, new collagen fibers thickened and assumed regular arrangement, and the collagen sponge films disappeared completely. In the control groups, the changes were less obvious and total HHK degradation occurred till week 12. Conclusion The degradable and absorbable HHK-collagen sponge film has relatively satisfactory tissue compatibility and can accelerate wound healing by stimulating cell proliferation and vascularization,showing the potential as an optimal dermal substitute.

AIMTo develop a rapid and feasible method based on micellar electrokinetic capillary chromatography (MECC) for the simultaneous determination of antiepileptic drugs (AEDs)--phenytoin (PHT), phenobarbital (PB), carbamazepine (CBZ), primidone (PRM) and clonazepam (CZP) in human plasma.

METHODSSeveral factors that impact the separation of AEDs with MECC were investigated, such as concentration of sodium dodecyl sulfate (SDS), buffer compositions, pH, organic modifier, internal diameter and temperature, and an optimized MECC running condition was obtained the running buffer consisted of 8 mmol x L(-1) phosphate, 3 mmol x L(-1) sodium tetraborate, and 50 mmol x L(-1) sodium dodecylsulfate (SDS) (pH 8.0), containing acetonitrile (ACN) (18%) as organic modifier. Detection at 210 nm, run at 25 kV at 30 degrees C in a untreated fused silica capillary (50/45.5 cm length, 50 microm ID).

RESULTSThe reproducibility of both migration time and relative peak area with MECC analysis were appropriate for the intra- and inter-assay coefficients. The evaluated drugs concentration intervals of PRM 1.0-40.0 microg x mL(-1), PB 1.0-60.0 microg x mL(-1), PHT 1.0-40.0 microg x mL(-1), CBZ 1.0-40.0 microg x mL(-1), CZP 0.2-8.0 microg x mL(-1) were linear with correlation coefficients higher than 0.999 1, and coefficients of the variation of the points of the calibration curve lower than 10%. The recoveries of AEDs varied from 80.0% to 100.0%, depending on the drug, with coefficients of the variation lower than 10.0%.

CONCLUSIONThe MECC technique is showed to be rapid, simple, efficient and low cost when applied to monitoring therapeutic drugs in patient treated with a combination of PHT and other AEDs such as hepatic enzyme-inducing agents.

Anticonvulsants ; blood ; Buffers ; Carbamazepine ; blood ; Chromatography, Micellar Electrokinetic Capillary ; methods ; Clonazepam ; blood ; Epilepsy ; blood ; Humans ; Hydrogen-Ion Concentration ; Phenobarbital ; blood ; Phenytoin ; blood ; Primidone ; blood ; Sensitivity and Specificity ; Sodium Dodecyl Sulfate