[Objective] To develop biomimicking chitosan scaffolds with longitudinally oriented micro-channels,and investigate their efficacy in bridging 15 mm sciatic nerve gap in rats.[Methods]Chitosan scaffolds with longitudinally oriented micro-channels were fabricated using unidirectional freezing-dry methods.The chitosan scaffolds were used to bridge 15 mm nerve defect in rats,and their efficacy in bridging nerve gap was evaluated by morphometric analysis,retrograde labeling,electrophysiological studies and behavioral analysis.[Results]The chitosan scaffolds developed in the present study showed longitudinally oriented micro-channels,which resembled the dimensions of basal lamina channels in normal nerves.Implantation of chitosan scaffold achieved axonal regeneration and functional recovery similar to autograft implantation in vivo.[Conclusion]The chitosan scaffolds have inner microstructures which resemble the basal lamina channels in normal nerves.The chitosan scaffold may be used as an alternative to autograft in bridging nerve gaps.

Objective To induce the site-directed mutation of human cardiac troponin I (cTnI) gene, express the mutant in E. coli, and to study the effects of the mutation on the prokaryotic expression of cTnI. Methods The cDNA encoding cTnI was cloned with RT-PCR from the total RNA extracted from human myocardium tissues. A pair of primers was designed and, after the mutations were induced at the second and the fourth codons, inserted into prokaryotic vector pET-28c (+) and transform the recombinant to BL21 (DE3) bacteria. After purified with Ni-NTA resin, the histidine-tagged fusion protein expressed by IPTG-induced was identified by Western blotting and the expression yield of cTnI protein was investigated. Results The expression of the recombinant carrying processed cTnI cDNA was stronger than that in control group. Conclusion cDNA encoding cTnI was successfully cloned. The recombinant with mutations can be more efficient expressed in E. coli. The cTnI protein can be purified to near homogeneity.

Comet Assay

Objective:To analyze the optimal application groups and influent factors of MMSE were studied. Methods:A total of 4921 old people aged 55 years and over were selected by means of randomized stratification cluster sampling from the urban and rural areas in Xi'an. The MMSE scores distribution and positive predictive values were analyzed among subjects of different ages and educational levels. Multiple stepwise regression method was used to evaluate significant variables. Results:MMSE scores showed a negative and cliff distributions for different ages, education levels, as well as for the whole sample. Moreover, these features were found to be most pronounced in relatively younger old subjects (less 70 years) with comparably higher educational levels (upper high school). Accordingly, the screening-out rates for dementia were higher among relatively older subjects with poor educational levels, when compared to younger and fairly well-educated subjects. Variables such as education, age, sex, hearing loss and daily functional abilities could significantly affect the MMSE score ( P

Objective:To identify the relationship between expression of protein kinase R(PKR), phosphating PKR, EIF2α(p-PKR, p-EIF2α) in PKR→EIF2α signal transduction passage and the grades of cervical lesions, the role in generation and progression of cervical tumor and their effects to prognosis of cervical cancer patients. Methods:The expressions of PKR, p-PKR and p-EIF2α in human cervical cancer tissue of 63 cases, cervical intraepithelial neoplasia(CINⅠ-Ⅲ) of 114 cases and normal cervical epithelium of 15 cases were detected by immunohistochemical technique. Results:With the increase in grades of cervical lesions, the positive-expression rate of PKR increased and significantly correlated with the grades of cervical lesions(P < 0.05). With the increase in grades of cervical lesions, the positive-expression rates of p-PKR and p-EIF2α increased firstly, and then decreased. In cervical cancer group, the positive-expression rate of PKR was much higher than that of p-PKR(P < 0.01). The development and progression was quicker in later clinical stages of cervical cancer than that of earlier clinical stages of cervical cancer (P < 0.01). The development and progression of cervical cancer was quicker in patients with negative-expression of p-PKR and p-EIF2α than that in patients with positive-expression of p-PKR and p-EIF2α(P < 0.05). Conclusion:The positive-expression rate of PKR was correlated with the grades of cervical lesions. There are some factors which can impede PKR and EIF2α to be phosphorylated or make p-PKR and p-EIF2α dephosphorylate in high level cervical lesions, which promotes the development and progression of cervical lesions, worsens the prognosis of cervical cancer.

After screening,the authors established a mouse model of pathomorphologically special,transplantable mammary spindle cell carcinoma SCC 891 in BALB/c mice.After 48 passages in 2.5 years,the growth of the tumor strain became stable with a 100% transplantibility.Its growth was quite active without observable spontaneous regression.The mean life span of the host mouse was 35.2?7.3 days.BALB/c mice of both sexes and of any age were susceptable to the carcinoma,and the carcinoma was sensitive to various anticancer drugs.

ObjectiveTo report the treatment of the first imported Middle East respiratory syndrome (MERS) in China, and to investigate the clinical features and treatment of the patient.Methods On May 28th, 2015, the first patient of imported MERS to China was admitted to Department of Critical Care Medicine of Huizhou Municipal Central Hospital. The clinical features and treatments of this patient were analyzed.Results①A 43 years old male of South Korean nationality was admitted with the complaint of back ache for 7 days and fever 2 days with the following characteristics: back ache 7 days ago, without fever or cough or expectoration. He had been suspected to suffer from infection of Middle East respiratory syndrome coronavirus (MERS-CoV) by the Disease Control Department of South Korea, but no specific treatment was given. He had fever for 2 days with maximum body temperature of 39.7℃. He had no chills, cough, expectoration, short of breath, abdominal pain, diarrhea, frequent micturition, or urgency or pain of urination, and no sore throat. The patient had a history of exposure to MERS-CoV patient. He was considered to be a patient of the second batch of South Korean epidemic.② Auxiliary examination: 3 copies of throat swab specimens for virus nucleic acid detection were performed by the Disease Prevention Control Center of China (China CDC), and they were positive on May 29th, 2015, and also for serum, sputum and stool. Based on the results of whole genome sequence analysis, the virus strains were implicated to be derived from Riyahh and Jeddah regions of Saudi Arabia. On admission, the patient's blood test showed that the white blood cell count was low (3.22×109/L), the proportion of the neutrophils was high (0.73), and that of the platelet was low (81×109/L). On admission, the patient's chest X-ray showed that a small amount of infiltration in the lung.③ Treatment: a high-flow nasal cannula (HFNC) with oxygen concentration of 0.50-0.80 was given, with a flow rate was set at 60 L/min if tolerated. It was changed to a low flow oxygen inhalation nasal cannula on the 20th day, and oxygen treatment was stopped on the 24th day. Ribavirin 2.0 g was given as the first dose, and was switched to 600 mg every 8 h (q8h), and it was reduced to 600 mg q12h after 10 days, and extenuated since the 13th day. Ceftriaxone was added on the 4th day with 2.0 g a day , and it was changed to meropenem 2.0 g, q8h on the 7th day for 2 weeks. Gamma globulin was given for 7 days (20 g, qd). Thymosin-α1 was given on the 8th day for 2 weeks. Interferon was given once a week, but only one dose was used. At the same time symptomatic treatment such as methimazole and liver protection therapy were given.④ Patient began to cough at admission, and it disappeared on the 18th day. There was no sputum at first, then a small amount of sputum with a little blood appeared after the admission. Then there was cough without sputum. Mild shortness of breath and diarrhea after exertion were noticed. He had no chest pain, difficulty in breathing or other symptoms. There was dullness on percussion in both sides of chest, and it disappeared gradually. Fine moist rales were detectable in scapular area and interscapular area on the 5th day, and they disappeared after 3 days. Breath sounds on both sides was weak, and it became more obvious in the right lung after 5 days, and returned to normal after 18 days. He had a sustaining fever for 1 week with the maximum temperature of 39.5℃, then the body temperature returned to normal. The viral nucleic acid test as performed by the Center for Disease Control of Guangdong (CDC, Guangdong) showed that the pharyngeal swab cultured turned negative on the 3rd day, that of serum specimens turned negative on the 8th day, that of stool specimen after 2 weeks, and it was persistently positive for sputum culture until 5 days before discharge. The oxygenation index gradually increased, and it was over 300 mmHg (1 mmHg = 0.133 kPa) after 15 days. Pleural effusion was rapidly increased during the first week as shown by chest X-ray films, and it began to be absorbed gradually in the second week, but it was not completely absorbed until discharge.Conclusions The disease course of the reported patient was short, with an acute onset, with fever as the chief complaint, but there were no respiratory symptoms, though there were high fever, cough, shortness of breath, diarrhea and other clinical symptoms after admission. Virus in sputum disappeared after treatment, but pleural effusion was not completely absorbed. Negative test for virus in sputum was late, indicating that clearance of virus was slow from the lungs. It is the first case of MERS in China, therefore, the clinical manifestations and the treatment strategy need to be further explored.

Infantile spasms constitutes an unique,age-specific epilepsy syndrome of early infancy characterized by epileptic spasms often accompanied by neurodevelopmental regression and an electroencephalograph finding of hypsarrhythmia.For the complex and various etiologies,the pathogenesis remains unclear.Although the relevant guidelines and consensuses are being updated all the time,but proven therapies are still limited because of efficacy,tolerability,at times even availability.In this paper,the current findings regarding up-to-date advances in the treatment of infant spasms were reviewed,especially in use of the corticosteroid and vigabatrin as well as the quality-of-care indicators for infantile spasms.

Objective To evaluate the therapeutic effect of adipose-derived mesenchymal stem cells on radiation enteritis.Methods A total of 52 male Sprague-Dawley rats were used in the present study.Herein,46 rats were randomly selected and irradiated with a dose of 15 Gy at their abdomens.Two hours post-irradiation,23 rats were randomly selected and infused intraperitoneally with adipose-derived mesenchymal stem cells in passage 6 from young-female donor.The other 23 rats were intraperitoneally infused with PBS.The rest 6 rats were set as normal control.During the first 10 days post-irradiation,peripheral blood-samples from irradiated rats were harvested for testing the levels of IL-10 in serum using ELISA assay.Additionally,after isolating the thymic cells and peripheral blood mononuclear cells,the percentages of CD4/CD25/Foxp(3)-positive regulatory T cells in thymus and peripheral blood were tested by flow-cytometry.Finally,infiltration of inflammatory cells and deposition of collagens within irradiated small intestine were analyzed by H&E staining and Masson Trichrome staining,respectively.Based on the MPO-immunohistochemistry staining,the type of infiltrated cells was identified.The Kaplan-Meier method was used for analyzing the survival rate of irradiated rats.Results During a period of 30 days post-irradiation,the irradiated rats receiving adipose-derived mesenchymal stem cells survived longer than those receiving PBS (t =4.53,P ＜ 0.05).Compared to the irradiated rats with PBS-treatment,adipose-derived mesenchymal stem cells could elevate the level of IL-10 in serum (7 d:t =13.93,P ＜ 0.05) and increase the percentages of CD4/CD25/Foxp(3)-positive regulatory T cells in both peripheral blood (3.5 d:t =7.72,7 d:t=11.11,10 d:t =6.99,P ＜0.05) and thymus (7 d:t =16.17,10 d:t =12.12,P＜ 0.05).Moreover,infiltration of inflammatory cells and deposition of collagens within irradiated small intestine were mitigated by adipose-derived mesenchymal stem cells.Conclusions Adipose-derived mesenchymal stem cells were capable of curing radiation enteritis.

Objective To assess the therapeutic effect of human adipose-derived mesenchymal stem cells on radiation-induced vascular injury in the small intestine of rat. Methods A total of 34 male Sprague-Dawley rats were enrolled in this study. To establish a model of radiation-induced intestinal injury, each rat was irradiated with 15 Gy in whole abdomen. 17 rats were randomly selected and infused intraperitoneally with passage 6 ( P6 ) Ad-MSCs, and the other 17 rats that received PBS were set as control. 10 days post-irradiation, the number of CD31+ endothelial cells in the small intestine villus was measured by flow-cytometry, the expressions of CD31, CD105 and isolectin-B4 in the na?ve endothelial cells with detected by IHC-staining, and the vascular integrity was evaluated by measuring VE-Cadherin. The origination of na?ve endothelial cells within injured intestine was also analyzed. In addition, total mRNA were extracted from irradiated small intestine to assay the expressions of VEGF, bFGF, Flk-1 and SDF-1 using quantitative Real-time PCR. Results Compared to the control, the amount of CD31-postive endothelial cells within irradiated intestine was significantly increased after Ad-MSCs infusion ( t=12?15, P<0?05). The microvascular density in the injured sites was also significantly increased by the infusion of Ad-MSCs (20 d:t=10?33, P<0. 05;30 d:t=32?85, P<0?05). Moreover, the expressions of VEGF, bFGF, Flk-1 and SDF-1 were significantly up-regulated after delivery of Ad-MSCs ( VEGF:t =10?34, bFGF:t=11?25,Flk-1:t=6?73, SDF-1:t=6?73, all P<0?05), which was beneficial in maintaining the integrity of intra-villus blood-vessels as well as promoting neovascularization in the injured sites. Conclusion Ad-MSCs had potentials in healing radiation-induced vascular injury in rat small intestine.