Objective To explore the differential expression of low molecular weight proteins in serum of ischemic stroke patients.Methods Serum samples obtained from ischemic stroke patients (n=33) and normal controls (n=39) were analyzed by Matrix Assisted Laser Desorption/ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS). The severity of stroke was assessed with the National Institutes of Health Stroke Scale (NIHSS) and functional outcome was assessed with modified Rankin Scale (mRS). Results 55 peaks were significantly different between ischemic stroke patients and the controls (P<0.05). There were 7 peaks between 1000~2000 Da, 4 increased peaks and 3 induced peaks in serum of ischemic stroke patients. One induced peak at 1864(m/z) had highest peak intensity in controls and induced significantly in ischemic stroke patients. While induce degree of this peak didn't correlate with the NIHSS, mRS, stroke risk factors and laboratory data. Conclusion The application of this potential biomarker is not restricted to certain subgroups of ischemic stroke patients,so it may serve as one reverse direction biomarker and provide support for early diagnosis and treatment of ischemic stroke.

Objective To establish a continuous monitoring assay of serum argininosuccinate lyase (ASL) activity with automatic biochemistry analyzer, and perform methodology validation and preliminary clinical application.Methods According to the chemical reaction catalyzed by ASL and the working characteristics of automatic biochemistry analyzer, an enzyme coupled reaction system with high specificity was set up, and the methodology validation was performed.Three hundred and nine patients with various liver diseases, 269 non-liver disease patients and 40 healthy controls were enrolled in this study.Serum ASL, ALT, and AST level were determined in all subjects.Results A new kinetics assay of ASL activity was set up with automatic biochemistry analyzer.The methodological validation demonstratod that inter-assay and intra-assay coefficient of variation were 4.0% and 5.9% respectively and the mean recovery was 100.5%.The linear range was 0-167.7 U/L.The lowest detection limit was approximately 0 U/L.The interference test showed that there is no significant interferences while the concentration of bilirubin is less than 342 μmoL/L or commonly used anticoagulants is employed at their routine concentrations.However,interference was significant when Hb level is more than 0.06 g/L.Preliminary study of clinical application showed that there was no significant difference of serum ASL level between non-liver disease group and healthy group ( q = 0.027, P = 0.979 ), but there was significant differences for both serum ALT and AST levels (ALT:q =6.461,P =0.000;AST:q =6.481,P =0.000).Conclusions A continuous monitoring assay for the determination of serum ASL activity is successfully established. Serum ASL may be a good biomarker for liver injury.

Objective To investigate the effect of cold atmospheric plasma (CAP) on the healing of skin ulcers using Balb/c mice. Methods Wounds with a diameter 6 mm were created on each side of the backs of BalB/c mice ( n = 150) using a punch bioptome. The mice were assigned randomly into a control group ( wounds healed naturally), a laser group (wounds treated with a He-Ne laser for 10 min daily) and a CAP group (wounds treated with CAP for 10 min daily). Wound healing was evaluated on postoperative days (PODs) 4, 7, 10 and 14 in terms of percent wound closure. Ten mice per group were sacrificed on each of the evaluation days. Both wounds were removed and a histological examination was conducted. A scoring system was used to evaluate the wounds. The expression of vascular endothelial growth factors (VEGFs) in the wounded tissue was detected by using immunohistochemical methods on POD 7. The results were quantified using an HPIAS-1000 system. Results Compared with the control group, the average percentage of wound healing was significantly greater in the CAP group on PODs 7 and 10. The average scores on the histological examination were significantly higher in the CAP group on PODs 7, 10 and 14. Compared with the other two groups, the expression of VEGF was up-regulated significantly in the CAP group.Conclusions CAP can positively affect the wound healing process. This might be related to the up-regulation of VEGF in the wounded tissues.

Objective To establish an appropriate animal model of brain metastases from lung cancer in nude mice by thoracic orthotopic implantation in the chest or left ventricular injection , and to serve further studies on the mechanisms of lung cancer brain metastasis .Methods PC-9 cells (1 ×106/0.1 mL) in logarithmic phase were respectively injected into 18 nude mice by orthotopic implantation in the chest or left ventricular injection ( n=9 each group ) .The statuses of nude mice were observed after implantation .Animals showing clear signs of dyscrasia were killed .At autopsy, the lung, brain, liver and kidney were removed and histological sections were stained with H /E to detect the presence of tumor cells . Results In the thoracic orthotopic implantation group , three weeks after implantation , the number 4, 6, 9 mice showed tumor nodules in the chest wall , they began to lose weight in the fourth to sixth week differently , showing signs of dyscrasia gradually , and were sacrificed at the fifth to seventh week .The thoracotomy revealed that the whole thorax was occupied by many large lung cancer masses , spreading into bilateral ribs , pleura and spinal vertebra , with scarce eroded , compressed , pale and distorted lung tissues left .Histological examination with HE staining showed the presence of neoplasms in their lung tissues but only the number 6 mouse showed metastatic lesions in the brain tissue .In the left ventricular injection group, the mice almost began to lose weight in the third week simultaneously and became moribund slowly , which were all sacrificed at the fourth week .After thoracotomy , the thoraxes were clear except the number 11 and 18 mice which appeared 2-3 tiny tumor foci in the chest wall , with normal lung tissues .Histological examination with HE staining showed the pres-ence of brain metastases in all the nine mice .The rate of brain metastases from lung cancer in the left ventricular injection group was 100%, compared with 11.1% in the thoracic orthotopic implantation group .Conclusions The establishment method of mouse model by left ventricular injection shows significantly higher rate of lung cancer brain metastases than that by thoracic orthotopic implantation .

Objective To identify and compare melanin distribution in iris and skin tissues of rabbits from four breeds including WHBE rabbit.Methods Ferrous sulphate staining was performed for iris and skin tissue of rabbits from different breeds.Results It showed dark green staining in both iris stroma and iris pigment epithelium of WHBE rabbit, while light green staining was observed in its follicles.Conclusion The results of histologic evaluations provide some clues to the future research on mechanism of WHBE rabbit phenotype mutation.

Objective To observe and explore the therapeutic effect of acupuncture combined rehabilitation training in treating with spasm cerebral palsy patients.Methods 128 spasm cerebral palsy patients in our hospital from February 2012 to May 2014 were selected and randomly divided into the observation group (68 cases) and the control group (60 cases).The patients of the control group were given massage,ganglioside intravenous treatment,the patients of the observation group were given acupuncture combined rehabilitation training on the basis of the control group.The total treatment was 3 months,after the treatment,the Ashworth score and the total effective effective rate of the two groups were compared.Results After treatment,the observation group of patients with Ashworth spasm rating scale significantly better than that of control group (P ＜ 0.05),the observation group of patients and the control group after treatment in patients with Ashworth spasm of rating scale were significantly better than that of before treatment (P ＜ 0.05),the treatment of patients with observation group total effective rate is significantly higher than control group (P＜ 0.05).Conclusions On the basis of the comprehensive treatment of acupuncture combined rehabilitation training therapy in patients with spastic type cerebral palsy treatment effect significantly,can significantly improve the patient' s level of limb muscle tension,worthy of clinical popularization and application.

Objective To analyze the pathogenesis,initially diagnosed symptoms and clinical manifestations of children with chronic kidney disease (CKD) at stage 2 to 5.Methods The data of 108 children who were hospitalized in Children's Hospital Affiliated to Capital Institute of Pediatrics from September 2007 to April 2016 with CKD stage 2 to 5 were retrospectively analyzed.The etiologies,clinical manifestations and examinations were summarized,and the clinical manifestations were compared between the congenital hereditary urinary diseases group and the acquired urinary diseases group.Results (1) In the 108 cases collected,66 cases were male,42 cases were female,aged from 3 months to 15 years and 1 month old.Twenty-four cases were diagnosed at stage 2,26 cases at stage 3,35 cases at stage 4,and 23 cases at stage 5.(2) Twenty-eight kinds of illness were involved in the cause of CKD.Among them,57 cases (52.8％) had congenital anomalies of the kidney and urinary tract,5 cases(4.6％) had hereditary kidney diseases,41 cases (38.0％) had other primary or secondary kidney diseases,and in 5 cases (4.6％) the causes of disease were unknown.(3) For the initially diagnosed symptoms,29 cases(26.9％) were due to complaints associated with kidney disease,36 cases (33.3％) were of other outside kidney symptoms,and 43 cases (39.8 ％) were of negative symptoms.The results of urinary ultrasound were abnormal in 79 cases(73.1％) and 87 cases(80.6％) showed abnormality in urinary analysis.There were 105 cases (97.2％) with abnormal manifestations either in urinary tract ultrasound or in urinary analysis.(4)The ages on diagnosis as CKD in children with congenital hereditary urinary diseases(5.89 years old) were younger than that of children with acquired urinary diseases (9.20 years old),and the difference was significant(Z =-3.434,P =0.001).The frequency of cases with short stature or lower-weight in group of congenital hereditary urinary diseases[66.1％ (41/622 cases),64.5％ (40/62 cases)] were significantly higher than those of the acquired urinary diseases group[43.9％ (18/41cases),43.9％ (18/41 cases)],and the differences were statistically significant(x2 =4.983,4.263,P =0.026,0.039).Conclusions The causes of CKD are complicated,and the congenital anomalies of kidney and urinary tract are the major causes of CKD at stage 2 to 5 in the cases.The initially diagnosed symptoms of CKD are insidious and atypical.The children with congenital hereditary urinary diseases tend to have more serious growth retardation.Urinary analysis and ultrasound may have an important significance for early diagnosis of CKD in children.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Proliferation ; Interleukin-10 ; genetics ; metabolism ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Organisms, Genetically Modified ; Rats ; Rats, Sprague-Dawley ; Transfection

Objective To observe the ultrastructure of blood-brain barrier in the nude mouse model of brain me-tastases from lung cancer by transmission electron microscopy using lanthanum nitrate tracing.Methods PC-9 cells (1 × 106/0.1 mL) in logarithmic phase were respectively injected into six nude mice ( model group) selected from eight nude mice randomly via the left ventricle, the other two mice without any treatment as the control group.The general status of the mice was observed after implantation.In the fourth week all the mice were sacrificed and brain tissue samples were taken and prepared for transmission electron microscopic observation using lanthanum nitrate tracing.besides, the lung and brain were removed and stained with HE to detect the presence of tumor metastasis.Results Mice in the model group began to lose weight almost simultaneously in the third week and became moribund slowly, and were all sacrificed at the fourth week when showing clear signs of cachexia.At autopsy, the thoraxes were clear, with normal lungs.Histology showed evidence of brain metastasis in all the six mice.The electron microscopy showed that lathanum nitrate tracer was escaped from the capillaries and diffusely or sparsely distributed in the brain tissues of the model group mice, however lathanum nitrate tracer was still confined in the capillary lumen in the mice of control group.Conclusions The diffuse lathanum nitrate tracer in the brain parenchymal tissue indicates the impairment of blood-brain barrier in the nude mouse model of lung cancer brain metastasis and the formation of these metastases is accompanied with the destruction of blood brain barrier.

Objective To evaluate the effects of progesterone on polymorphonuclear leukocyte (PMN)-mediated inflammatory responses to gonococcal infection. Methods Peripheral neutrophils were isolated from heparinized peripheral blood obtained from normal individuals, then divided into 4 groups: progesterone group (pretreated with progesterone only), gonococcus group (stimulated with gonococcal suspension), intervention group (pretreated with progesterone followed by stimuation with gonococcal suspension), and control group (receiving no pretreatment or stimulation). Real-time RT-PCR was conducted to detect the mRNA expression levels of inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β)in neutrophils from all groups at 0, 3, 8, 12 and 24 hours after the last treatment, and iNOS protein levels were measured by Western-blot in gonococcus group and intervention group. Results Real-time RT-PCR indicated that the expression levels of iNOS, TNF-α and IL-1β mRNA increased in gonococcus group and intervention group, and reached their peak at 8 hours in gonococcus group, while no significant changes were noted in the above parameters in progesterone group or control group. Also, the level of iNOS, TNF-α and IL-1β mRNA was lower in intervention group than that in the gonococcus group (P ＜ 0.05). Western blot showed an elevation in iNOS protein expression in both gonococcus group and intervention group, and the former group was higher than the latter group in the parameter (P ＜ 0.05). Conclusions Progesterone can downregulate the expressions of iNOS, TNF-α and IL-1 β by PMNs, inhibit the PMN-induced inflammatory responses induced by gonococcal infection, which is likely to be associated with the asymptomatic gonococcal infection in women.