BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)are widely utilized as seed cells or carriers in bone tissue engineering and gene therapy.Thus,how to obtain BMSCs with high purity arose more attentions of researchers.OBJECTIVE:To observe the effects of different separation methods and culture conditions on biological characteristics of rabbit BMSCs.METHODS:BMSCs were obtained by whole bone marrow culture,density grand centrifugal and red blood cell lysis.At 48 hours after culture,the cell numbers were counted,the time of passage was recorded,in addition,the cell morphology was observed by phase contrast microscope,and the CD44 antigen expression was identified using flow cytometry.The 3rd and the 7th generation aging cells were cultured with DMEM-LG,MEM-HG,and DMEM/F12 culture medium.MTT and count cell plat were used to evaluate the growth of BMSCs.Phase contrast microscope was used to observe the morphological changes of aging cells.RESULTS AND CONCLUSION:BMSCs could be separated by each method.The adherent cells showed shuttle or multiple angle shapes,with rich cytoplasm,and positive for CD44 antigen.The more cell number and shortest primary culture time was presented in red blood cell lysis group(P ＜ 0.05).DMEM/F12 could promote the proliferation of quiescent cells.And the cells prevented the better viability.The method of read blood cell lysis improving the efficiency of BMSCs adherent is an effective method of extraction of BMSCs.DMEM/F12 could promote the proliferation Maybe,DMEM/F12 is more suitable for BMSCs.

Objective To investigate the relation between the changes d myocardial collagen content before and after ventricu-lar assist device (LVAD) application and the prognosis in dilated cardiomyopathy. Methods Patients was divided to two groups ac-cording to the prognosis (weaning or transplantation) after LVAD application. Left ventricular samples were collected from the patients (weaning group n= 10, transplanted group n=8) at the time the LVADs were implanted, and again during cardiac transplantation ( n = 8 ). The content of neutral salt soluble collagen (NSC) and acid soluble collagen (ASC) were measured by Sired collagen assay and total collagen and insoluble collagen (ISC) by quantification of hydroxyproline. Moreover, protein quantification of each collagen fraction was performed simultaneously. Results Before LVAD implantation all collagen fraction and their protein contents were higher in the weaning group than that in the transplanted group ( P <0.05), but the difference in the collagen fraction disappeared after LVAD support. Comparison of the pre- and post-LVAD subgroups of the transplanted group showed an increase of NSC and its protein content and total hydroxyproline after LVAD support (P<0.05). Conclusion The significances of myocardial intersititial remodeling in DCM are not identical. Some myocardial collagen and protein content increases may be related to good prognosis. The mechanism of LVAD improving cardiac function may be related to the changes of myocardial intersitial collagen.

To compare the adjuvant therapeutic effect of silicone oil and ointment applied to the lacrimal laser angioplasty.METHODS: Totally, 134 eyes of 102 cases who were diagnosed as lacrimal duct obstruction ( except chronic dacryocystitis) were divided into two groups. Sixty eyes were performed laser plastic operation and silicone oil filling. Seventy - four eyes were adopted laser plastic operation with cloth enzyme dexamethasone eye ointment filling. All patients were followed up for 3 ~ 15mo. The clinical effect and the number of irrigation of two groups were compared 3mo after treatment, and to assess the merits of the method.RESULTS: The efficiency of the silicone oil group was 87% and that of the eye ointment group was 84%. There was no statistically significant difference (P>0. 05). The number of lacrimal duct flushing of the silicone oil group was 7. 20±1. 22 times among postoperative 3mo, and that of the eye ointment group was 7. 96 ± 1. 79 times. The number of lacrimal duct exploration through of the silicone oil group was 0. 62 ± 0. 87 times, that of the eye ointment group was 1. 43 ± 1. 48 times. There was statistically significant differences compared between the two groups (both P<0. 05).CONCLUSlON:The effect of laser plastic operation and silicone oil filling treating obstructive diseases of lacrimal duct is similar to laser plastic operation with cloth enzyme dexamethasone eye ointment, but the postoperative care of the former is more simple. Patients can significantly reduce the frequency of lacrimal duct flushing and lacrimal duct exploration through.

Advance Care Planning (ACP) was introduced as part of patient care in the acute hospitals in Singapore in 2010, and has been gaining traction among hospital providers and their patients. The Agency for Integrated Care (AIC) hopes to strengthen ACP training and awareness in the social and community care sectors. Primary care physicians play an important role to introduce ACP, provide relevant information based on patients health status, and offer advice, encouragement and guidance to enhance understanding, reflection and discussion. They can help patients and their loved ones identify issues in coping with chronic diseases or possible life-threatening conditions. When done well, ACP can uphold the respect and dignity of patients and allow for patient-centric care continuation. ACP conversation often begins by listening and exploring the patients story. Physicians should be attuned to their patients fears, worries and concerns, and identify triggers for opportunistic discussions about ACP with them. With adequate ACP and communication training, primary care physicians are probably best placed for timely ACP initiation.

Objective To investigate the clinical characteristics of neonatal tuberous sclerosis complex (TSC).Methods A total of 134 patients were admitted and diagnosed as TSC in Beijing Children′s Hospital,Capital Medical University from September 2006 to September 2015.The clinical characteristics of TSC in 8 patients who had clinical symptoms initiating from the neonatal period (skin lesion,neurologic abnormality,etc.) were analyzed,so as to conduct the auxiliary examinations (skin biopsy,ultrasonic cardiogram,cranial imaging) and the follow-up results.Results Among 8 patients,4 were diagnosed as TSC in the neonatal period while other 4 were diagnosed at later period from 4 months to 14 years old.Six patients had skin lesions in neonatal period,accounting for 75% of 8 patients,with predominant symptoms:hypomelanotic macules (5 cases),angiofibroma (2 cases),and shagreen patch (1 case).Cardiac abnormalities in neonatal period mainly included cardiac rhabdomyoma,which were characterized by high incidence,multi-regional occurrence,and wide distribution,noticed in 4 patients (50%).Ultrasonic cardiogram showed a mass in 4 patients with moderate-strong echo,uniform texture and clear boundary,and slight effect on tricuspid valve blood flows (in 1 case).Electrocardiogram abnormalities were found in 1 case,like atrial premature beats with intraventricular aberrant conduction,and accelerated atrial escape.Neurologic abnormality of neonatal TSC could present convulsive seizures.Cranial imaging lesions showed subependymal nodules (3 cases),leukodystrophy (2 cases),and giant-cell astrocytoma (1 case).Evidence for TSC2 gene positive and heterozygous mutation was identified in 1 case.The nucleic acid mutation site was at c.268C>T (E4),and the amino acid mutation was p.90Q>X.The mutant effect was nonsense mutation,which could lead to premature termination of protein translation.Conclusion The clinical characteristics of neonatal TSC may involve multi-system lesions,most commonly seen in the skin,cardiac,and ner-vous system.Neonatal physical examinations,cranial CT/magnetic resonance imaging and cardiac ultrasound screening should be done for the suspected TSC patients,and genetic diagnosis may contribute to the early diagnosis of the disease.

ObjectiveTo identify the probability for outdoor walking independency using the Berg Balance Scale (BBS) and the maximum walking speed （MWS） in stroke patients. Methods80 hemiparetic stroke patients were classified into 2 groups: the independency group and the non-independency group. They were assessed with the BBS and the 10 m MWS. ResultsWith the discriminant analysis, the BBS and the 10 m MWS could identify the independency and non-independency group，hit rate 88.8%. Receiver operating characteristic (ROC) curves suggested the cut-off point 49.5, sensitivity 86.2%; specificity 78.4%, area under ROC curve 0.905 in BBS; while cut-off point 1.0 m/s, sensitivity 93.1%, specificity 90.2%, area under ROC curve 0.961 in MWS. ConclusionBBS and the MWS can identify the outdoor walking independency sensitively and specifically in stroke patients.

Objective To explore the high-risk factors of the abdominal wound undesirable healing in gynecologic operations and its prevention and treatment.Methods A retrospective clinical study reviewed 58 cases in gynecologic operations.To investigate and analyze its frequent clinical risk factors.Results The risk factors in these wound healing defect such as the fat liquefied:3 cases(48.3%),the anaemia and/or hypoproteinemia:13 cases(22.4%);combined with diabetes:3 cases(5.2%),hypertension:6 cases(10.3%),wound infection:2 cases(3.5%)and wound hematoma:1 case(1.7%);two high risk factors coexisted:22 cases(37.9%),three or more risk factors coexisted:11 cases(19.0%).The complication of such wound healing defect such as effusion:26 cases(44.8%),flare and induration:28 cases(48.3%),low-grade fever:5 cases(8.6%).Conclusion The fat liquefaction is the important reason of the wound dehiscence in gynecology operations;the effusion,flare or induration is the precipitation of the wound healing defect.If we can treat these high-risk groups positively,we will improve the clinical prognosis.

BACKGROUND: Cytokine is a poly-functional and effective regulator factor to regulate growth of multiple cells. Researches suggest that, as an inflammatory medium, cytokines play a key role in inflammatory reaction of eye; however, there are rare studies on dynamic changes after traumatic cataract.OBJECTIVE: To investigate the relationship among inflammatory reaction and dynamic changes of interleukin 1 (IL-1),IL-6 and tumor necrosis factor alpha (TNF-α) in aqueous humor of rabbits after extracapsular cataract extraction.DESIGN: Randomized controlled animal study.SETTING: Shenzhen Municipal Ophthalmology Hospital, Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University.MATERIALS: The experiment was carried out in Institute of Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University from March 2004 to March 2006. Fifteen healthy adult New Zealand rabbits of 30 eyes,general grade, weighting 2.5-3.0 kg, of either gender, were provided by Animal Center of the Second Clinical Medical College of Jinan University [certification: SYXK (yue) 2005-006]. Eyes of rabbits were normal before experiment. All rabbits were randomly divided into normal control group, traumatic control group and operative group with 5 in each group and in total of 10 eyes in each group. IL-1, IL-6 and TNF-α kits were provided by Shenzhen Yawei Biotechnology Company Limited.METHODS: Rabbits in traumatic control group and operative group were totally anesthetized with intravenous injection of 10% 1 mi/kg urethan, and then, 5# needle was punctured from corneal limbus to anterior chamber to scarify anterior membrane of lens about 5 mm to establish animal models of traumatic cataract of oculus uterque. Rabbits in normal control group were fed normally. After successful modeling, common antibiotic eyedrops was used to clean conjunctival sac of rabbits in traumatic control group and operative group 3 times a day. On the 3rd day of successful modeling, rabbits in operative group were totally anesthetized with intravenous injection of 10% 1 mL/kg urethan, and then, they undertook extracapsular cataract extraction of oculus uterque: horizontally intercepting bladder or waterly separating with breakage of anterior bladder membrane, expulsing nucleus of lens, washing lens cortex and suturing incisions. On the operative day and on the 1st, 3rd, 7th and 14th days after operation, inflammatory reaction of anterior chamber in traumatic control group and operative group was measured and 0.2 mL aqueous humor was extracted from rabbits in three groups to count and classify cells; meanwhile, expressed level and dynamic changes of IL-1, IL-6 and TNF-α in cytokines of aqueous humor were measured with double-antibodies ELASA technique.MAIN OUTCOME MEASURES: Total numbors of leucocytes and contents of IL-1, IL-6 and TNF-α in aqueous humor of rabbits in three groups after operation.RESULTS: ① On the 1st, 3rd, 7th and 14th days after operation, numbers of leucocytes were (2.4±0.7)×106/L, (2,2±0.5)×106/L, (2.8±0.8)×106/L and (2.0±0.5)×106/L in aqueous humor; (19.7±7.3)×106/L, (28.1±9.6)×106/L, (14.2±5.6)×106/L and(8.4±3.8)×106/L in traumatic control group; (65.3±14.5)×106/L, (79.8±12.7)×106/L, (21.7±8.2)×106/L and (12.4±4.1)×106/L in operative group. In addition, numbers of leucocytes were more in traumatic control group and operative group than those in normal control group (F =22.5, 27.9, 11.6, 8.4;P＜0.05). ② Within 1-14 days after operation, contents of IL-1, IL-6and TNF-α in aqueous humo were higher in traumatic control group and operative group than those in normal control group (P＜0.05), and there was a significant difference between traumatic control group and operative group (P＜0.05);however, there was no significant difference among three groups on the operative day (P＞0.05). ③ Contents of cytokines reached peak on the 7th day after operation, decreased gradually, and reached the lowest value on the 14th day.Contents in traumatic control group and operative group were higher than those in normal control group (P＜0.05).CONCLUSION: The intraocular inflammation after lens extraction is closely related to the dynamic changes of IL-1, IL-6and TNF-α levels in aqueous humor. Cytokine may be one of crucially inflammatory agents in the eyes after traumatic cataract.

Objective To evaluate the effect of health education on preventing birth defects in community in terms of knowledge, attitude and practice (KAP) for married women in reproductive age. Method A self-designed general information form and a questionnaire on KAP of married women′s preventing birth defects were used to investigate the KAP among 300 married nulliparous women. Result The average rate of cognitive knowledge of birth defects was 60.8%, the average rate of cognitive behaviors was 59.2%, and the average rate of cognitive attitude was 67.4%. Conclusion In order to reduce the rate of birth defects and improve the quality of births, we should implement three-level prevention education system, enrich the health education methods to enhance the knowledge, attitude and behavior of married women.

The article discusses reforming information search course, integrating the education of users training and information search course for training the persons with ability of innovation or scientific research in new century, strengthening the combination of theory and practice, reforming the way of exams, emboding the individuation in teaching amd strengthening the ability of information analysis and catch .