BACKGROUND:Adrenomedulin gene transfection can strength the anti-apoptotic ability of bone marrow mesenchymal stem cels under ischemia and hypoxia, but its mechanism is not yet clear.
OBJECTIVE:To investigate the effect of adrenomedulin on the expression of apoptosis-related proteins, Bax, Bcl-2 and Caspase-3, in bone marrow mesenchymal stem cels under hypoxia and ischemia.
METHODS:Bone marrow mesenchymal stem cels of Sprague-Dawley rats were isolated, cultured and purified, and then cultured in serum-free medium under hypoxic condition for 0, 3, 6, 9, 12 hours. Then, western blot assay was employed to detect the expression of Bax, Bcl-2 and Caspase-3 so as to determine the optimal hypoxia time that was determined at 6 hours of hypoxia. Depending on whether adrenomedulin pretreatment was done, the cels were divided into control group (with no adrenomedulin pretreatment before hypoxia and ischemia) and adrenomedulin groups with different concentrations (1, 10, 100 μg/L). Afterwards, the expression of Bax, Bcl-2 and Caspase-3 was detected by using western blot assay.
RESULTS AND CONCLUSION:(1) After cultured in serum-free medium under hypoxia for 0, 3, 6, 9, 12 hours, the expression of Bax, Bcl-2 and Caspase-3 in bone marrow mesenchymal stem cels were increased (P < 0.05);at 6 hours of hypoxia, the Bax/Bcl-2 ratio and Caspase-3 expression reached the minimum value (P < 0.05). (2) At 6 hours of hypoxia, the expression of Bax and Caspase-3 protein as wel as Bax/Bcl-2 ratio became the lowest in the 100 μg/L group compared with the 1 and 10 μg/L groups, but the expression of Bcl-2 protein reached the peak (P < 0.05). These findings indicate that adrenomedulin can reduce the expression of Bax/Bcl-2 ratio and Caspase-3 protein in bone marrow mesenchymal stem cels cultured in serum-free medium under hypoxic conditions, which is in a dose-dependent manner.

BACKGROUND:Previous studies have demonstrated that hepatocyte growth factor (HGF) gene transfection can improve the effectiveness of bone marrow mesenchymal stem cel transplantation, but the mechanism is stil unclear. OBJECTIVE:To observe the effects of HGF gene transfection on c-MET, Bax, Bcl-2, Caspase-3 of bone marrow mesenchymal stem cel s cultured under hypoxia and serum-free conditions. METHODS:(1) Bone marrow mesenchymal stem cel s were isolated and amplified in vitro by differential adhesion method. The infection efficiency of recombinant adenovirus Ad-HGF in bone marrow mesechymal stem cel s was tested by x-gal staining. (2) Bone marrow mesenchymal stem cel s were cultured under hypoxia and serum-free conditions for 0, 3, 6, 9, 12 hours. RT-PCR and western blot assays were used to evaluate the expression of Bax, Bcl-2, Caspase-3. (3) Bone marrow mesenchymal stem cel s were cultured under hypoxia and serum-free conditions for 6 hours, and RT-PCR and western blot assays were adopted to detect HGF, c-Met, Bax, Bcl-2 and Caspase-3. (4) Cel scratch test was used to detect the effect of HGF transfection on the migration of bone marrow mesenchymal stem cel s cultured under hypoxia and serum-free conditions for 6 hours. RESULTS AND CONCLUSION:(1) Transfection efficiency of bone marrow mesenchymal stem cel s was increased with multiplicity of infection in a dose-dependent manner. When the multiplicity of infection was 150, the transfection efficiency was 96.4%. (2) Expressions of Bax and Bcl-2 were gradual y increased with hypoxia time (P<0.05). The Bax/Bcl-2 ratio and Caspase-3 expression reach the minimum at 6 hours of hypoxia (P<0.05). (3) Compared with the control and Ad-LacZ groups, the expressions of HGF, c-Met, Bcl-2 increased, and the expressions of Bax and Caspase-3 decreased in the Ad-HGF group after 6 hours of culture under hypoxia and serum-free conditions (P<0.05). There was no significant difference between the control and Ad-LacZ groups. (4) The mobility of bone marrow mesenchymal stem cel s was higher in the Ad-HGF group than the control group and Ad-LacZ groups after 6 hours of culture under hypoxia (P<0.05). These findings indicate that transfection of HGF in bone marrow mesenchymal stem cel s can increase the expression of c-Met, Bcl-2 and decrease the expression of Bax, Caspase-3 under hypoxia and serum-free conditions, which also enhance the mobility of bone marrow mesenchymal stem cel s under hypoxia and serum-free conditions.

Objective To explore the relationship between the characteristics of dynamic contrastenhanced MRI (DCE-MRI) and the stages of endometrial cancer.Methods A retrospective analysis of DCE-MRI manifestation in 55 patients with scratch-pathologically confirmed endometrial cancer was performed.All patients were divided into different groups according to differentiation,muscle layer infiltration,cervix involvement and metastasis.The enhanced rate,including the arterial phase relative signal increase ratio (ARSI％),the maximal relative signal increase ratio (MRSI％) and the signal enhancement ratio (SER％),was calculated at different time (i.e.16 s,32 s,48 s,64 s and 300 s),respectively.Time-intensity curves (TIC) of lesions were also obtained and divided into 4 subtypes.Two sample t test was performed to compare the MRSI％ among groups,and the Mann-Whitney U test was performed in ARSI％ and SER％ as well.Results The early phase enhanced rate of normal tissue was 894 ± 355,higher than that of tumor 716 ± 215 (t =-2.911,P ＜ 0.01).The delayed phase enhanced rate of normal tissue was 1111 ± 289,higher than that of tumor 803-± 289 (t =-4.926,P ＜ 0.01).ARSI％ of low differentiation group (n =16,M =1.07％) was higher than that of middle to high differentiation group (n =39,M=0.36％).ARSI％ of deep muscle layer infiltration group (n =19,M =0.76％) was higher than that of no muscle layer infiltration group (n =36,M =0.32％).ARSI％ of cervix involvement group (n =27,M =0.84％) was higher than that of no cervix involvement group (n =28,M =0.25％).SER％ of cervix involvement group (n =27,M =2.90％) was higher than that of no involvement group (n =28,M =1.40％) and SER％ of parauterine involvement and metastasis group (n =7,M =2.03％) was higher than that of uterine involvement group (n =48,M =1.60％).The differences between these groups were statistically significant (Z =-2.038,-2.260,-2.172,-2.695 and-2.621 respectively,P ＜ 0.05).Based on the TIC types,type Ⅰ occurred in 13 tumors (23.6％),type Ⅱ in 22 tumors (40％),type Ⅲ in 9 tumors (16.4％),and type Ⅳ in 11 tumors (20％).Conclusions Based on TIC curves,most of endometrial cancers showed significant enhancement which reflected of their rich blood supply.The parameters of DCE-MRI is helpful in staging of uterus tumors.

Objective To discover radioresistance-associated proteins by performing comparative proteomic analysis on nasopharyngeal carcinoma cell lines.Methods The total proteins were extracted from radioresistant human nasopharyngeal carcinoma cell line CNE-2R and its parental cell line CNE-2,respectively.These proteins were separated by high quality two-dimensional polyacrylamide gel electrophoresis (2-DE) and then the 2-DE profiles were screened for differentially expressed protein spots by the Image Master 5.0 software.Those spots were identified by a matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry.Results 32 significantly differentially expressed protein spots were screened in two different radiosensitivity cell lines and 11 proteins were identified by tandem mass spectrometry,among which 3 proteins were up-regulated in radioresistant human nasopharyngcal carcinoma cell line CNE-2R and the other 8 proteins were down-regulated.Conclusions The differentially expressed proteins of nasopharyegeal carcinoma cells with different radiosensitivity were mainly involved in apoptosis regulation,DNA damage and repair,cell cycle regulation,RNA transcription,cell signaling,cytoskeleton formation and radiation stress responses.

Objective To study the influence of obesity on outcome of rehabilitation for the hemiplegic post stroke. Methods 118 hospitalized stroke patients were divided into the normal weight, overweight and obesity groups according to the Body Mass Index. All the patients received rehabilitation for 12 weeks. They were assessed with National Institute of Health Stroke Scale (NIHSS), Fugl-Meyer Assessment (FMA), Berg Balance Scale (BBS) and modified Barthel Index (MBI) before and after rehabilitation. Results The scores of NIHSS, FMA, BBS and MBI improved significantly after rehabilitation in all the groups (P<0.001), and improved the lest in the obesity group in the scores of NIHSS, BBS and MBI (P<0.05). There was no significant difference in FMA score among groups (P>0.05). Conclusion Obesity influences the outcome of rehabilitation, results in poor functioning.

Objective:To evaluate the accuracy of parents ’ perception of whether their child is a picky eater and the specific food category the children avoideating according to the food frequency question-naire.Methods:This research selected 1 663 infants aged 4-36 months receiving non-diary complimen-tary food from maternal, infants, nutrition and growth study (MING Study) in 8 Chinese cities in which a combination of systematic cluster random sampling and purposive sampling was used .The general infor-mation, dietary status and picky eating status were collected through a self-designed questionnaire from the caregiver of the children .According to the parents ’ perception , the children were classified into picky/non-picky groups or avoid/non-avoid to a specific food category groups .Mann-Whitney U test was used to compare between the groups .Results: The reported percentage of picky eaters increased from 7.37%in 4-6 months old infants to 36.20%in 25-36 months old infants .Most picky infants in 4-6 months and 7-12 months old infants avoided eating dairy food ( 25% and 24%); while most picky toddlers aged 13-24 months and 25-36 months avoided eating vegetables (26.92% and 47.46%). The infants aged 4-6 months and 7-12 months who were perceived as picky by their parents took more kinds of food (8 and 19.5 kinds) than those who were not (6 and 18 kinds), while the picky toddlers aged 13-24 months and 25-36 months took fewer kinds of food (28.5 and 34 kinds for picky eaters, 31 and 37 kinds for non-picky eaters ) .The parents of infants aged 4-6 months judged correctly in every category of food without any statistical significance;the parents of 7-12 months old infants judged cor-rectly only in dairy food and eggs with statistical significance;those of 13-24 months old infants judged correctly in every food category except for vegetables with statistically significant difference in the category of eggs;those of 25-36 months old toddlers misjudged in dairy , beans and grains with no statistically significant difference in every category .Conclusion: Parents tend to misjudge their children ’ s picky eating behavior before the first 12 months of the child , and tend to make a more accurate perception after the 12th month.

By summarizing the policies and regulations of foreign countries on rare disease research and drug research and development, referring to the relevant policy support of treatment guarantee system in the development of rare diseases abroad, and combining with China’s national conditions, this paper analyzed the feasible points that China can learn from, and then put forward some suggestions on the ethical problems existing in the current rare disease research, so as to provide reference suggestions for the relevant policy formulation of rare disease treatment guarantee in China: adjusting regional differences based on the principle of fairness, and establishing a national rare disease information collection center; improving the ethical quality of scientific research in pharmaceutical companies, and strengthening ethical supervision; and the government and society fulfill their obligations to protect the rights and interests of patients with rare diseases.

There are a large number of patients with rare diseases in China, and most of them are faced with problems such as no medicine to cure, and have drugs outside of China but not inside. This paper combed the incentive policies of orphan drugs in the United States and the European Union, including orphan drugs legislation and setting up special management institutions, orphan drugs qualification certification, government funding, tax reduction and agreement assistance in the research and development process, providing accelerated listing channels in the examination and approval process, giving market monopoly period in the circulation stage, and giving priority to review. On this basis, it also explored the incentive measures of non-profit organizations in the research and development of orphan drugs, including providing financial support, clinical research and so on. From the perspective of guaranteeing the right to life and health of patients with rare diseases, the fairness and accessibility of medication, and the subjects’ right to know and privacy in clinical trials, this paper provides reference for perfecting the incentive mechanism of orphan drugs research and development in China.