There are many ways to extract and culture neural stem cells in vitro, but the viability and stability of neural stem cells obtained by different methods are different. By thinking about the process of extracting and culturing neural stem cells in vitro from the cerebral cortex of SD fetal rats, we summarized extraction steps, the main points of extraction, the selection basis of culture medium, selection of inoculation density, cultural method, methods of solution changing, passage time and passage methods. At last a large number of neural stem cells with high vitality and stability have been obtained and applied to the basic research of neural stem cells.

ObjectiveTo investigate the effect of capsaicin on cognitive dysfunction in rats with cerebral ischemia-reperfusion and its possible mechanism. MethodTwelve SD male rats were randomly selected as a sham operation group， and the remaining rats were sutured to replicate the model of middle cerebral artery occlusion （MCAO）. The successfully modeled rats were divided into a model group， a SB203580 ［p38 mitogen-activated protein kinase （p38 MAPK） inhibitor， 1 mg·kg^-1］ group， capsaicin low- and high-dose （50， 100 mg·kg^-1） groups ， and anisomycin （p38 MAPK agonist， 2 mg·kg^-1） + capsaicin （100 mg·kg^-1） group， with 12 rats in each group. After reperfusion and administration， the rats were scored for neurological deficits. Morris water maze and new object recognition experiments were used to test the learning and cognitive abilities of rats. The hematoxylin-eosin （HE） staining was used to observe the pathological changes in the hippocampus of the brain tissue. Immunofluorescence method was used to detect the activation of microglia in the hippocampus. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and prostaglandin E₂ （PGE₂） inflammatory factors in the brain tissue. Western blot was used to determine the protein expression levels of transient receptor potential vanillin subfamily 1 （TRPV1）， p38 MAPK， p-p38 MAPK， and cyclooxygenase-2 （COX-2） in the hippocampal tissue. ResultAs compared with the sham group， the neurological deficit score， escape latency， the number of Iba-1 positive microglia in the hippocampal CA1 area， the IL-1β， TNF-α， and PGE₂ levels in the brain tissue， and the p-p38 MAPK/p38 MAPK and COX-2 expression in the hippocampus tissue was significantly increased in the model group （P<0.01）. In the model group， the number of crossing the platform position， the novel object discrimination index （DI）， and the TRPV1 expression in the hippocampus tissue was significantly reduced （P<0.01）， the number of hippocampal nerve cells was reduced， and a large number of inflammatory cells infiltrated. As compared with the model group， the neurological deficit score， escape latency， the number of Iba-1 positive microglia in the hippocampal CA1 area， the IL-1β， TNF-α， and PGE₂ levels in the hippocampus tissue， and the p-p38 MAPK/p38 MAPK and COX-2 expression in the hippocampus tissue were significantly reduced in the capsaicin low-dose and high-dose groups （P<0.05，P<0.01）. In the capsaicin low-dose and high-dose groups， the number of crossing the platform position， the DI， and the TRPV1 expression in the hippocampus tissue were significantly increased （P<0.05，P<0.01）， a small amount of inflammatory cells were infiltrated， and the number of nerve cells was significantly increased. The use of anisomycin， an activator of p38 MAPK， increased the expression of COX-2， and significantly weakened the inhibitory effect of capsaicin on the activation of microglia. ConclusionCapsaicin has a protective effect on the cognitive function of rats with cerebral ischemia-reperfusion， and its mechanism may be related to the inhibition of the activation of p38 MAPK/COX-2 signaling pathway， thereby inhibiting the excessive activation of microglia.

OBJECTIVE:To observe clinical efficacy and safety of recombinant human endostatin(rh-endostatin)com-bined with CT-guided percutaneous microwave ablation in the treatment of non-small cell lung cancer(NSCLC)complicat-ed with chronic obstructive pulmonary disease (COPD). METHODS:A total of 80 cases of NSCLC complicated with COPD were selected from our hospital during Feb. 2014-Feb. 2016,and then divided into control group and observation group according to random number table,with 40 cases in each group. Control group was treated by CT-guided percutane-ous microwave ablation. Observation group was additionally given rh-endostatin injection 7.5 mg/m2,once a day,d1-14, added into 500 mL 0.9％ sodium chloride injection,ivgtt lasting for 4 h,for consecutive 14 d,on the basis of control group;7 d later,next course was performed. A treatment course lasted for 21 d,and they received 4 courses of treat-ment. Survival time,clinical efficacy as well as KPS score and lung function indexes before and after treatment,the oc-currence of ADR were compared between 2 groups. RESULTS:Median survival time of observation group(19.8 months) was significantly longer than that of control group(15.2 months),and total response rate of observation group(72.5％) was significantly higher than that of control group (55.0％),with statistical significance (P<0.05). Before treatment, there was no statistical significance in KPS score,FEV1,FEV1/FVC or DLCO between 2 groups(P>0.05). After treat-ment,KPS score and above lung function indexes levels of 2 groups were increased significantly,and those of observa-tion group were significantly higher than those of control group,with statistical significance (P<0.05). No grade Ⅳwas found in 2 groups. There was no statistical significance in the cases of ADR at different levels between 2 groups (P>0.05). CONCLUSIONS:rh-endostatin combined with CT-guided percutaneous microwave ablation in the treatment of NSCLC complicated with COPD show good clinical efficacy with less ADR,and can significantly improve lung function and quality of life.

With the continuous advancement of health informatization and the wide application of medical big data, electronic health records came into being and spread rapidly. However, because electronic health records contain a large amount of private information, privacy protection is the primary consideration for the sustainable development of electronic health records. By analyzing the shortcomings of privacy protection of electronic health records in law, technology, management and protection consciousness, this paper put forward some countermeasures, such as perfecting the relevant laws and regulations of privacy protection of electronic health records, improving the technical level, improving the management defects of electronic health records, and cultivating the privacy protection consciousness of professionals and the public, so as to improve the overall privacy protection level of China’s health records information management system and provide effective protection for the privacy information of Chinese residents’ electronic health records.

OBJECTIVE: To observe the clinical effect and safety of acupuncture in treatment of neck pain due to cervical spondylosis. METHODS: According to the patients' preference and acceptance for the interventions of neck pain induced by cervical spondylosis, an acupuncture group (221 cases) and a non-acupuncture group (251 cases) were divided. After the control of confounding factors with propensity score matching, 218 cases were included in either acupuncture group or non-acupuncture group. In the acupuncture group, acupuncture was applied to Dazhui (GV 14), Baihui (GV 20), ashi points, bilateral neck-Jiaji (EX-B 2), Fengchi (GB 20), Houxi (SI 3), Shenmai (BL 62), etc. The treatment was given once daily, one course of intervention was composed of 5 treatments and 3 courses were included. In the non-acupuncture group, the oral administration of imrecoxib tablets and cobalt tablets was prescribed for 2 weeks. Before and after treatment, the scores of Northwick Park questionnaire (NPQ) and the simplified McGill pain questionnaire (SF-MPQ) were observed, and the safety was assessed in patients of the two groups. RESULTS: After treatment completion, the scores of NPQ and SF-MPQ were all reduced when compared with those before treatment in each group (P<0.001), and the scores of NPQ and SF-MPQ in the acupuncture group were lower than those of the non-acupuncture group (P<0.001). The incidence of adverse reactions was 6.0% (13/218) in the acupuncture group and was 10.1% (22/218) in the non-acupuncture group, without statistical significance in comparison (P>0.05). CONCLUSION Acupuncture is effective and safe in the relief of neck pain and the improvement of comprehensive quality of life in the patients with cervical spondylosis.
Humans ; Neck Pain/therapy* ; Propensity Score ; Quality of Life ; Acupuncture Points ; Acupuncture Therapy ; Spondylosis/therapy* ; Treatment Outcome

Objective To investigate the differentiated thyroid carcinoma diagnosis and treatment options. Methods From Feb. 2002 to Jan. 2008,78 patients received different surgical resection regarding the type of tumor size,number of tumor,ages and jugular lymphatic metastasis. Patients with unilateral differentiated thyroid carcinoma underwent the resection of ipsilateral isthmus of thyroid lobe or plus partial contralateral gland,and those with bilateral-lobe underwent total thyroidectomy or near-total thyroidectomy. High-risk patients (age >45 years,tumor size >4 cm,tumor size ≤4 cm,but surpass the envelop of thyroid) were performed by functional neck dissection or lymph node dissection of central region (Ⅵ area) besides postoperative endocrine therapy. Results Eleven cases underwent the resection of ipsilateral lobe with isthmus, 19 cases underwent surgical removal of ipsilateral lobe with isthmus plus partial contralateral gland,26 cases underwent near-total thyroidectomy and 22 total thyroidectomy. 25 cases underwent functional neck dissection, 23 cases underwent neck dissection of central region. There were 68 papillary thyroid carcinoma (87. 18%), 10 follicular thyroid carcinoma (12. 82%). There were 26 cases with lymphatic metastasis of Ⅵ area. Postoperative complications included 12 cases (15.38%) with deadlimb caused by hypocalcemia, 8 cases (10.26%) with transient recurrent nerve paralysis,2 cases (2.56%) with permanent injury of recurrent laryngeal nerves, 3 cases (3. 58%) with chylous fistula. Seventy-four(94. 87%) cases were followed up postoperatively for a period from 6 months to 6 years,which showed that no death occurred,but 6 relapsed with jugular lymphatic metastasis,after reoperation no distant metastasis occurred. Survival rate was 97. 30% (72/74). Conclusion Treatment of the differentiated thyroid carcinoma should be based on the size of tumor,number of tumor,age and jugular lymphatic metastasis. Lymph node dissection of central region was necessary for high-risk patients of differentiated thyroid carcinoma.

To improve the part of the hardware about Xin Hua SL-IC model X-ray radiotherapy simulative position finder, using CR and PACS, so that located-picture shooting for the simulative position finder have been realized digital upgrade.
Automatic Data Processing ; Radiographic Image Enhancement ; Radiotherapy ; methods ; Tomography, Spiral Computed ; methods

OBJE CTIVE To establish a method for quality evaluation of Xin ’an capsule by combining fingerprint ， multi-component quantitative analysis and chemical pattern recognition analysis. METHODS High performance liquid chromatography（HPLC）combined with Similarity Evaluation System of TCM Chromatogram Fingerprint （2012 edition）were used to establish the fingerprints of 24 batches of Xin ’an capsules and evaluate the similarity. The common peaks were determined. The contents of glucosylvitexin ，rhamnosylvitexin，vitexin，hyperoside and isoquercetin in Xin ’an capsules were determined by the same HPLC method. Taking the common peak area of fingerprint as the variable ，MetaboAnalyst 5.0 tool was used to draw the cluster analysis （CA）heat map. SIMCA 14.1 software was used to perform principle component analysis （PCA）and partial least squares-discriminant analysis （PLS-DA）. RESULTS Twelve common peaks were identified with the similarity greater than 0.97. Six common peaks were identified as chlorogenic acid ，glucosylvitexin，rhamnosylvitexin，vitexin，hyperoside and isoquercetin.The linear range of glucosylvitexin ，rhamnosylvitexin，vitexin， hyperoside and isoquercetin were 2.36-151.35，9.15-585.20， 1.20-76.50， 0.68-43.20， 0.44-27.90 µg/mL（all r＞0.999）.RSDs of precision ，repeatability and stability （24 h）tests were 163.com all less than 2.00％ . The average recoveries were 95.80％（RSD＝0.96％ ，n＝6），102.10％ （RSD＝0.93％ ，n＝6）， 103.26％（RSD＝1.28％，n＝6），103.89％（RSD＝0.73％，n＝6） and 102.09％（RSD＝1.79％，n＝6），respectively. The contents of the five components were 0.988 8-1.559 1，4.336 6-11.220 1， 0.065 1-0.830 5，0.043 8-0.692 5 and 0.023 2-0.427 2 mg/grain，respectively. The results of CA and PCA showed that 24 batches of samples could be divided into three categories ，i.e. S 1-S15，S16-S18 and S 19-S24. PLS-DA showed that variable importance in projection values of the corresponding component of peak 6 and glucosylvitexin （peak 7），rhamnosylvitexin（peak 8），hyperoside （peak 10） and isoquercetin （peak 11） were greater than 1. CONCLUSIONS The established HPLC fingerprint and multi-component quantitative method are simple and feasible. Combined with chemical pattern recognition analysis ，it can be used for the quality control of Xin ’an capsules. Glucosylvitexin ，rhamnosylvitexin and other components may be differentital markers affecting the quality of each batch of samples.

Objective To look for a tissue paraffin embedding method that can replace xylene transparent agent. Methods The mixture of N-butanol and turpentine was used to replace the role of anhydrous ethanol and xylene in the process of conventional paraffin embedding. The brain, kidney, stomach, liver and duodenum of multiple cerebral infarction model rats were embedded in paraffin. Finally, the new dehydration procedure was evaluated according to the effect of paraffin section, HE staining and immunohistochemical result . Results The mixture of N-butanol and turpentine could replace not only the dehydration effect of anhydrous ethanol but also the transparency effect of xylene in the conventional paraffin embedding process. The tissue sections treated with the mixture of N-butanol and turpentine were smooth, and the tissue did not become brittle or hard; After HE staining, the nucleus and cytoplasm of the new dehydrated tissue were distinct, and the chromaticity, color and transparency of the tissue were not different from those of the conventional dehydration procedure; Immunohistochemical staining was performed on different tissues of rats, and the comparison result were no different from conventional embedded tissue immunohistochemical staining. Conclusion N-butanol combined with turpentine for tissue dehydration can not only avoid the toxic effect of xylene on human, but also reduce the tissue damage caused by excessive dehydration of anhydrous alcohol.

OBJECTIVE：To optimize the inclusion technology of volatile oil from Ganmao qingre granules. METHODS ： Guided by the concept of “quality by design ”，taking volatile oil inclusion rate and inclusion complex yield as key quality attribute，comprehensive score of above two indexes after weighting as response value ，inclusion temperature ，inclusion time ，the ratio of β-CD to volatile oil as key technology parameters ，Box-Burman response surface design was adopted to establish the design space of key technology parameters and key quality attributes. The design space was optimized and verified by 95％ confidence interval. The stability of inclusion complex was investigated preliminarily. RESULTS ：The optimal design space ，i. e. the optimal technology parameters rang ，included inclusion temperature 35-40 ℃，inclusion time 1.8-2.0 h，the ratio of β-CD to volatile oil 9.5∶1- 10∶1（g/mL）. The results of 3 validation tests showed that the volatile oil inclusion rates were all over 62％，the yields of inclusion complex were all over 75％，and the comprehensive scores were all over 80 point. The results of preliminary stability showed that the inclusion rate of volatile oil ，the yield of inclusion complex and the comprehensive score did not change significantly. The difference in evaluation indicators within 7 days was within 5％. CONCLUSIONS ：The optimized inclusion technology is feasible ， and the obtained inclusion complex is stable.