ObjectiveTo investigate the role of Wnt5a in the mechanism of radiculopathy and the relation between Wnt5a and tumor necrosis factorα(TNF-α) by observing the change of the expression of Wnt5a in the rat model of chronic compression of dorsal root ganglia (CCD). MethodsA total of 192 adult male Sprague Dawley rats were allocated into 4 groups: shame group (group A, n=48), CCD group (group B, n=48), CCD+sal ine group (group C, n=48), and CCD+etanercept group (group D, n=48). An L-shaped needle (about 3.5 mm in length, 0.6 mm in diameter) was inserted into the L5 intervertebral foramen, and the dorsal root ganglia (DRG) was compressed by the needle to prepare the CCD model in groups B, C, and D, and then normal sal ine (5.5 mg/kg) or etanercept was injected intraperitoneally in groups C and D. The intervertebral foramen was exposed in group A. The mechanical pain threshold of the posterior paw was tested by the von Frey filaments at 1, 3, 5, and 7 days after operation; the expressions of Wnt5a protein and mRNA were detected at 3 and 7 days after operation by immunohistochemical staining and RT-PCR, respectively. ResultsThe mechanical pain threshold of groups B and C was significantly lower than that of groups A and D, and in group D than in group A (P < 0.05), but no significant difference was found between groups B and C (P > 0.05). The Wnt5a positive cells and the mRNA expression of Wnt5a at 7 days were significantly more than those at 3 days in groups B, C, and D (P < 0.05). The Wnt5a positive cells and the mRNA expression of Wnt5a in groups B and C were significantly more than in groups A and D, and in group D than in group A (P < 0.05), but no significant difference was shown between groups B and C (P > 0.05). ConclusionThe expression of Wnt5a in the DRG is increased after CCD. The expression of Wnt5a in the DRG is decreased after the administration of the inhibitor of TNF-α.
Objective To investigate the synergetic effect and possibil ity of repairing spinal cord injury (SCI) by transplantation of olfactory ensheathing cells (OECs) and chondroitinase ABC (ChABC) in adult rats. Methods Three adult male SD rats were used to isolated olfactory bulb and primarily cultured OECs. In the 8th or 9th day, OECs were transplanted, the concentration of cells was modulated to 1 × 105/μL. Fifty-four SD rats were made the models of T8 spinal cord crush injury and divided into 4 groups. In group A (control, n=36), injured site was not treated; in groups B, C and D (n=6), OECs, ChABC and OECs+ChABC were injected into injured site, respectively. At 1, 2, 3, 7 and 14 days after injury, the BBB score system was used to evaluate the motion function. At 0, 1, 2, 3, 7, 14 days in group A and at 14 days in groups B, C, D after injury, the maximal transverse diameter and gross area of necrosis were evaluated on HE stained sections. The immunofluorescence double label ing staining for gl ial-fibrillary acidic protein (GFAP)/CS56, GFAP/growth associated protein 43(GAP-43) and GFAP/neurofilament 160(NF160) was carried out to evaluate the regeneration of nerve fiber. Results At 14 days after injury, there were significant difference in the BBB scores between group A and groups B, C, D (P lt; 0.05), and between groups B, C and group D (P lt; 0.05), HE staining showed that the formation of cavity was observed in each group at 14 days after injury. There were significant difference in the maximal transverse diameter and gross area of necrosis between groups B, C, D and group A (P lt; 0.01), and between groups B, C and group D (P lt; 0.01). The immunofluorescence staining indicated that expression of GFAP were more intense in group A than in other groups, and the cavity of the lesion site was apparent, but it was moderate in groups B and C. The expression of GAP-43 was more intense in group D than in groups B and C. The expression of NF160 was more intense in group D. Conclusion Transplantation strategy of OECs combined with ChABC was effective in the repair of SCI in some extent.
Objective To establish an animal model for repairing the sciatic nerve defect with a biodegradable poly D,L-lactic acid/nerve growth factor (PDLLA/NGF) that can control the release conduit in rats and to observe an effect of the conduit on the sciatic nerve regeneration. Methods The PDLLA conduit and the PDLLA/NGF-controlled release conduit (NGF 450 U per conduit) were madewith the solvent-volatilixation method. Forty male SD rats were randomly and equally divided into 4 groups. The middle segments (10 mm) of the sciatic nerves of the rats were excised and were then repaired with the sciatic nerve autograft(Group A), with the PDLLA conduit (Group B), with the PDLLA conduit and an injection of NGF (30 U) into the conduit (Group C), and with the PDLLA/NGF controlled-release conduit (Group D), respectively, with the 10-mm nerve defect left behind. Three months after operation, the morphologic parameters of the nerve regeneration were observed and evaluated under light microscope and electron microscope, and the image analysis was also made. Results Three months after operation, porous adherence between the conduit and the surrounding tissues could be observed. The conduit presented a partial biodegradation but still remainedintact in the outline and the proximal nerve regenerated through the conduit cavity. Based on the histological observation, the quantity, uniformity, and maturity of the nerve fiber regeneration in Groups A and D were better than those in Groups B and C. The image analysis indicated that there were no significant differences in the nerve fiber diameter, axon diameter or myelin thickness between Group A and Group D (P＞0.05). However, all the parameters in Groups A and D were better than those in Groups B and C (P＜0.05). Conclusion The PDLLA/NGF-controlled release conduit can effectively promote the sciatic nerve regeneration of rats. Its morphological index is similar to that of the nerve autograft.
Objective To establish a better method of isolating andculturing ofneural stem cells(NSCs) in neonatal rat brain. Methods Tissue of brain was isolated from neonatal rats. Different medium and culture concentration were used toculture NSCs of neonatal rat. The culture concentration used were 1×10 4, 1×105, 1×106and 1×107/ml respectively. Ingredient of medium was classified into group 1 to 8 respectively according to whether to add 2% B27, epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) as well as the difference in culture concentration. The cells were induced to differentiate asto be confirmed as NSCs, and then were checked by phase contrast microscopy and identified by immunocytochemistry. Results The cells isolated and cultured gathered into neurospheres. The cells were capable of proliferating and maintaining longterm survival in vitro. The cells could be differentiated into neurons and glia.It was to the benefit of the survival of NSCs to add 5% fetal bovine serum(FBS)into the medium at the beginning of the culturing. When 10% FBS was added intothe medium, the neurospheres differentiated quickly. When concentration 1×106/ ml was used, the growth rate of the cells was the highest of all the concentrations. Reasonably higher cell concentration promoted the proliferation of NSCs. It was necessary to add 2% B27, EGF, and bFGF into the medium. The cells had the best growth when 2% B27, 20 ng/ml bFGF and 20 ng/ml EGF were added into the culture medium. EGF and bFGF had cooperative effect. Conclusion A better method of isolating and culturing of NSCs in neonatal rat brain is established and the foundation for future research is laid.
Objective To explore the serious situation of injection abuse, and its influence to average prescription fee. Methods The subjects of this study were health service settings in rural area of 9 provinces/cities in Midwest of China. The treatment prescription indicators of county and village health service settings were calculated. Results Prescription injection rates of health care facility in rural area of Midwest provinces/cities of China (25.8% to 62.2%,mean: 45.1%) were higher than the standard of WHO (13.4% to 24.1%), and the injection abuse situation was serious. Injection bause caused the increase of prescription fee. Excess usage of injection in health service settings was related to the economic level of the on-site county or village, and also related to the size and load of health service facilities. Conclusion Suggestions are proposed to the government health agency according to the results of the study: enhancing the lawmaking, establishing the related policy and effective measure, training the medical personnel, promoting the mass health education, investigating the effective injection management model in rural area, and reducing the rate of injection.
Liver transplantation in hamster-to-rat is a good model for the research in xenograft, but it is difficult to be performed. In order to simplify its procedures, 38 cases of liver transplantation in hamster-to-rat were performed with several technique improvments on the basis of orthotopic liver transplantation in rats. In the light of hamster’s anatomy, three cuffs anastomosis method was used. Because of its high stability and high survival rate, the model can be used widely as the research for liver xenograft.
【Abstract】Objective To investigate the effect of donor blood transfusion on inducing pancreatic allograft tolerance in outbred rat model. Methods Wistar male rats were used as blood and pancreas donor, and diabetic recipients. One ml of donor blood injected into abdomen of diabetic recipients on the day of transplantation and azathioprine given 2 days pretransplant and continued for three days. Results Pancreas allograft survival was significantly prolonged (28 to 112 days, media survival time 64.2 days). One ml of donor blood alone injected into the abdomen and azathioprine given alone 2 days pretransplant did not improve allograft survival (media survival time 9.8 vs 10.2 days). Conclusion Donor blood injected on the day of transplantation and a 3 days course of azathioprine started 2 days pretransplant have b synergism in inducing long term graft survival in this rat model.
ObjectiveTo evaluate the effects of N-acetylcysteine (NAC) on lung tissue of Wistar rats, which were tracheally instilled fine particulate matter (PM2.5).MethodsForty-eight male Wistar rats were randomly divided into six groups: two control groups [they were blank group (C1), fake treatment group (C2) separately], four treatment groups [they were PM2.5 group (P), low-dose NAC group (L), medium-dose NAC group (M), high-dose NAC group (H) separately]. C1 received no treatments at all. C2 was instilled with sterile water (1 ml/kg) tracheally once a week for four times. P was instilled equivoluminal PM2.5 suspension (7.5 mg/kg) tracheally once a week for four times. The NAC groups received gavage (10 ml/kg) of different dosage of NAC (125, 250, 500 mg/kg) for six days. At the seventh day, the NAC groups were instilled PM2.5 suspension (7.5 mg/kg) tracheally. The procedures were repeated for three times in the NAC groups. Twenty-four hours later after four weeks or after the last instilling, all rats were sacrificed. Lung tissue was stained by hematoxylin-eosin (HE) staining, and histopathological changes of lung tissue were observed by optical microscope. The levels of C-reactive protein (CRP) as well as tumor necrosis factor-α (TNF-α) of serum, TNF-α of bronchoalveolar lavage fluid (BALF), TNF-α as well as interleukin-1β (IL-1β) of homogenates of lung tissue were detected by enzyme-linked immunosorbent assay. The activity of lactate dehydrogenase (LDH) as well as the levels of malondialhyde (MDA) of serum and BALF were detected by standard colorimetric method.ResultsHE staining showed that the normal structure of lung were destroyed in the groups dealed with PM2.5 and NAC could alleviate these changes. Higher dosage of NAC seemed to provide more powerful protections. Structure of the lung in C1 as well as C2 were nearly normal. The levels of CRP as well as TNF-α of serum, TNF-α of BALF, TNF-α as well as IL-1β of homogenates of lung tissue in the groups of P, L, M, H were higher than that in the groups of C1, C2 (all P<0.05). The levels of CRP as well as TNF-α of serum, TNF-α of BALF, TNF-α as well as IL-1β of homogenates of lung tissue in the groups of L, M, H which groups received NAC treatments were lower than that in P group. More, the groups seemed to have lower levels of CRP, TNF-α, IL-1β when higher dosage of NAC were given. The activity of LDH as well as the levels of MDA of serum, and BALF in the groups of P, L, M, H were higher than that in the groups of C1, C2 (all P<0.05). The activity of LDH as well as the levels of MDA of serum and BALF in the groups of L, M, H which groups received NAC treatments were lower than that in P group (all P<0.05). ConlusionTo some extent, NAC demonstrate antagonistic effects on oxidative stress and inflammatory injury on rats’ lung brought by PM2.5.
After the reform of drugs and medical consumables, the problem of unreasonable medical examination has gradually become prominent, which has become the potential threat of doctor-patient disputes and rising medical costs. At present, the reform of the medical and health system has entered the deep water zone. In the process of standardized management of medical examination, the difficulties in doctor-patient trust, medical examination supervision, medical examination resource allocation, the implementation of the mutual recognition system of medical results and the guarantee of information security have gradually emerged. In view of the dilemma of medical examination control, this paper proposes the following three breakthrough paths: based on the perspective of doctors and patients, improve doctor-patient trust; coordinating institutional supervision and administrative supervision to create transparent medical examinations; technical support to promote the standardized operation of medical examination, in order to provide a theoretical basis for the standardized operation of medical examination.
The current quantitative methods of bilirubin have disadvantages such as high cost and low sensitivity. Due to the negative correlation between the level of serum bilirubin and the risk of cardiovascular diseases, a fluorescent ratiometric film sensor was developed aiming at bilirubin detection at low level concentration. Blue-emitting and red-emitting gold nanoclusters were assembled into the same film using layer-by-layer self-assembly technology. Detection of bilirubin was achieved based on the intensity ratio of the two nanoclusters. Bilirubin exposure causes fluorescent quenching of the film. The fluorescence intensity ratio of the two cluster probes had quantitative relationship versus bilirubin concentration. Based on this film sensor, a portable fluorescence detection system was designed for the ratiometric sensing of bilirubin. The hardware of the system was mainly composed of main control chip STM32F407, TSL237 and TSL238T optical frequency sensor. A light-avoiding dark room and detection light path were designed through three-dimensional printing to reduce the interference from ambient light and improve detection accuracy. Experimental results showed that the proposed detection system had strong anti-interference, good stability and accuracy. The linear coefficient of bilirubin detected by this system was 0.987. The system presented good results in reproducible experiments and possessed a good linear relationship with the data obtained by standard spectrofluorometer. The portable system is expected to detect serum bilirubin at low levels.