An“agar-island organ culture”technique is described.It consists of both agar substratum and liquid medium in the same culture dish,so that chemicals in different concentrations can be added into the liquid portion of the medium,a convenient method to test their effects on explants.The procedures are as follows: To one gram of agar in a flask,50 ml of aqua distillata is added,and then heated until the agar is completely dissolved.Then add in 50 ml of double-strength 199 solu- tio(?) into the flask.After shaking gently,mixing the media completely,pour it into a dish until it reaches 2/3 of its height.A few minutes later the agar will solidify.Cut off most of the agar substratum and leave enough agar to form“islands”(Eig.A,B).Then add in liguid medium which is composed of 8 vol 199 solution,2 vol calf serum,2 vol extract of 9-day chick embryo diluted into 50% in Hank's solution and a few drops of diluted penicillin and streptomycin.The explants are placed on the agar-islands,the culture dishes covered with glass tops,sealed with paraffin,are put into an incubator.The cul- ture dishes are not sealed they are put into an incubator supplemented with 5%CO_2 Using this technique,we have cultured 80 explanls of 13-day chick embryo meta- tarsal skin for 7～21 days.And the effect of vit.A with different concentrations are tested The results show that the effects of vit,A on the differentiation of the chick embryo metatarsal skin explants are different with the different concentrations.We believe that this technique can be much more convenient as a method to study the eff- ects of chemicals in different concentrations on explants.The other uses of this tech(?)i- que,such as carcinogenesis in culture,the study of embryonic induction,especially the diffuse mechanism of embryo induction are also briefly discussed.

The present study reported the effect of two male antifertility agents gossypol acetic acid and GTW on DNA of C3H10T1/2 mouse fibroblasts. Our results showed that the cells treated with gossypol or GTW at high concentration (2-3 ?g/ml) for 4 hours, show silver grains in their nuclei as much as the positive control group, N-methyl N′-nitro-N-nitrosoguanidine (MNNG) a known carcinogen. However, if the agents were used at moderate concentrations (0.5-1?g/ml), the silver grains were much less, if the concentrations of gossypol or GTW were of 0.1-0.3 ?g/ml, the silver grains were as less as the control group. In a colony-forming test, we found that the cells lost their proliferate ability, since no colonies could be formed, if gossypol or GTW were of high concentration; while at moderate or low concentrations, the colony-forming rate was as high as 8.1-10.5％. Taking all of these results into consideration, we suggest that high concentrations of gossypol or GTW can damage cell DNA severely, moderate concentration of the agents break cell DNA to a certain extent, but the cells can repair, while low concentration of gossypol or GTW exert no obvious effect on cells. The significance of these observations was briefly discussed.

ObjectiveTo investigate the features of the cortisol ( COR),thyroid hormone and autonomic function of anxiety disorder,and to observe the relationships between clinical subtypes,insomnia,gender factors and the neuroendocrine and autonomic function.Methods60 anxiety disorder patients and 60 healthy controls were randomly selected from a three-grade class A hospital over the same period.Blood at 8:00am to test the plasma COR,T3,T4,TSH level and examined the heart rate variability (HRV) on the same day,the anxiety disorder patients also finish the self-rating anxiety scale (SAS).ResultsThe plasma COR,TSH level and LF/HF index were significantly higher( (531.58 ± 218.59 ) nmol/L,P＜ 0.0l ; (3.50 ± 1.35 ) MIU/L,P＜ 0.05 ;3.87 ± 3.37,P＜0.0l ),while the SDNN,LF,HF index were significantly lower in anxiety disorder than that in the control group (27.34±13.13,P＜0.01;338.72 ±396.34,P＜0.01;135.77 ±149.95,P＜0.01).The plasma COR,TSH level and LF/HF index were significantly higher(P ＜ 0.05,P＜0.01,P＜0.01 ),while the SDNN and HF index were significantly lower in panic disorder than that in the generalized anxiety disorder (P ＜ 0.01 ).The plasma COR,TSH level and LF/HF index were significantly higher,while the SDNN,LF,HF index were significantly lower in panic disorder than that in the control group (P＜0.01 ).The plasma COR was significantly higher,while the SDNN and HF index were significantly lower in generalized anxiety disorder than that of the control group (P＜0.01 ).The plasma COR level and LF/HF index were significantly higher (P＜0.01,P＜0.05),while the SDNN,LF,HF index were significantly lower in anxiety disorder with or without insomnia than that in the control group (P＜ 0.01 ).The plasma TSH level was significantly higher in anxiety disorder with insomnia than that in the control group (P＜ 0.05 ).There were no significant difference of the neuroendocrine and autonomic function between anxiety disorder with or without insomnia (P ＞ 0.05 ).The plasma COR,TSH level were significantly higher,while the SDNN,LF,HF index were significantly lower in the female group than that in the male group (P＜0.01).The total SAS scores were significantly positively correhted with plasma COR levels,and was significantly negatively correlated with SDNN,HF index in the anxiety group.ConclusionThere are neuroendocrine and autonomic nervous system dysfunctions in anxiety disorder,and these dysfunctions are related with clinical subtypes,insomnia,and gender.

Objective To establish a rapid genotyping method of for methicillin-resistant Staphylococcus aureus(MRSA) based on polymerase chain reaction(PCR)-high resolution melting (HRM ) curve analysis and staphylococcal protein A (SPA ) classifica-tion .Methods 71 strains of MRSA clinically isolated were collected as test strains .Gene sequencing and HRM curve analysis were employed to conduct SPA gene typing .Results According to gene sequencing method ,SPA gene of 71 strains of MRSA was divided into four types ,namely t570 ,t030 ,t002 and t588 .The most predominant type was t570 (74 .65% ) ,followed by t030 and t002(both 7 cases) .The result of SPA gene typing by HRM analysis were basically consistent with that by gene sequencing .Con-clusion PCR-HRM analysis is expected to become a fast ,efficient genotyping for MRSA SPA gene ,providing the basis for hospital infection control .

Objective To report the integrated progress of the hydatid disease information management system,and to pro-vide the reference for further system improvements by analysis of results on simulation test feedback. Methods The work of in-stitutional code matching by collecting fundamental and integrated information of the system in epidemic areas of hydatid disease was carried out,and professional control agencies were selected to carry out the simulation test. Results The results of agen-cies code matching at stage indicated the average completion rate was 94.30% on administrative agencies,69.94% on registered professional agencies and 56.40% on professional institutions matching related to hydatid disease prevention and control imple-ments in seven provinces(autonomous regions)and Xinjiang Production and Construction Corps. Meanwhile,the response rate of open-ended proposals was 93.33% on fifteen feedbacks,and the statistics showed 21.43% believed the system was low fluen-cy,64.29% considered the system was inconvenience for data inputs and 42.86% considered it would be improved on system sta-tistics functions,of which 27.78% were provincial users,22.22% were the city users and 50.00% were the county users. Con-clusion The hydatid disease prevention information management system meets the fundamental needs of the majority agencies in hyperendemic areas of echinococcosis,it needs to develop the further test with more agencies joining after the work of the in-stitutional code matching completion and the system service improvement in the next stage.

Objective To investigate the tempo?spatial patterns of schistosomiasis in Jiangling County,Hubei Province,so as to identify the risk areas and provide the scientific evidence in following intervention plans for marshland epidemic areas in the stage of transmission control. Methods The schistosomiasis epidemiological data in Jiangling County from 2009 to 2013 to?gether with the related geographical information were collected and analyzed. The tempo?spatial distribution patterns were ana?lyzed by the spatial autocorrelation analysis and spatial clustering analysis. Results The human infection rate was decreased from 2.15%in 2009 to 0.63%in 2013,which was the historically low level. The results of tempo?spatial analysis showed that there were spatial clustering effects in human schistosomiasis infection for each of the years. The values of spatial autocorrelation index Moran’s I were statistically significant. Eighteen and thirty?five clusters were detected by using SatScan and FlexScan soft?ware,respectively. Conclusions From 2009 to 2013,the schistosomiasis endemic situation in Jiangling County presented a decline trend and reached the historical low level. The identified spatial clustering areas should be targeted as the prioritized ar?eas for schistosomiasis control.

Objective To evaluate the feasibility and efficacy of atlantoaxial fusion by flipping periosteum pedicle occipital outer plate.Methods Between March 2010 and June 2013,27 patients with atlantoaxial instability were treated by atlantoaxial fusion with flipping periosteum pedicle occipital outer plate and combining pedicle screws fixation.There were 16 males and 11 females with the age ranging from 23 to 56 years (with an average of 45.6 years) at time of surgery.There were 12 cases of fresh odontoid fracture with atlantoaxial instability,7 cases of old odontoid fracture combined with uneducable atlantoaxial dislocation,7 cases of congenital loose of odontoid process,and 1 case of rheumatoid arthritis.The visual analog scale (VAS) scores and Frankel grades were respectively used to evaluate the axial neck pain and the neurological deficit,and the results were compared before and after the operation.Bony fusion was observed by the midline sagittal CT scan images.Results All the patients were successfully operated.The operation time was 90 to 140 mins (with an average of 100 mins),and the blood loss was 100 to 600 ml (with an average of 160 ml).All patients were followed up for 18 to 39 months,with an average of 32.6 months.19 cases got solid fusion 3 months after surgery,and 7 cases got solid fusion 6 months after surgery,while 1 case got solid fusion 9 months after surgery.The average VAS score at final follow-up was 2.87± 1.03,which was significantly lower than that preoperatively 7.23± 1.65.Before the operation,nine cases presented spinal nerve function damage.At the final follow up time,7 cases returned to complete normal and 2 cases improved from grade C to grade D according to the Frankel classification.1 case was complicated with veiniplex injury with no heavy blood loss,due to successful hemostasia.Two screws were placed close to the vertebral artery canal,without clinical consequences.Conclusion The flipping periosteum pedicle occipital outer plate as a bone graft is one effective method for atlantoaxial fusion,which got great feasibility and safety.

Objective To study the risk factors of intracranial hemorrhage after endovascular therapy of cere-bral arteriovenous malformations embolization.Methods the clinical records and images of 230 patients with cAVMs were retrospectively reviewed.Patients with intracranial hemorrhage after endovascular therapy in 3 days were recorded and the risk factors of hemorrthage by using multivariate logistic regression for guiding the treatment strategy were studied.Results There were 15 patients occurred intracerebral hemorrhages.After treatment the area of cAVMs was significantly smaller than that before treatment[(2.18 ±0.91)cm vs (6.67 ±1.56)cm],the difference lvas a statis-tically significant (t =9.627,P <0.05).A partial AVM reduction of ≥36.5%(P =0.031)and hypertension(P =0.025)were considered as risk factors for hemorrhage after endovascular therapy of cAVWs.Conclusion Patients receives a partial AVMreduction of ≥36.5% or with hypertension history have a tendancy of increasing hemorrhage after endovascular therapy of cAVMs.It's helpful for preventing intracerebral hemorrhage if the arterial blood pressure maintains after endovascular therapy.

The aim of this study is to develop a rapid and accurately species typing method for Brucella isolates by using High Resolution Melting (HRM ) analysis .Six pairs of primers were used according to the reference for the sequence of pur‐pose gene .Nineteen biotypes of six species Brucella standard strains were identified by PCR‐HRM analysis and this analysis was used to detect the 35 clinical isolates .Results showed Brucella amplified specific melting curves were different from con‐trasted strains with primer Bspp .The six species Brucella standard strains have own characteristic curve shape from each oth‐ers by PCR‐HRM analysis with five pairs of primers .Thirty‐five clinical isolates of Brucella have entirely consistent with PCR‐HRM curve shape with Brucella melitensis standard strains .So ,PCR‐HRM analysis methods can accurately identify Brucella strains ,especially clinical isolated Brucella melitensis ,and may be used in clinical microbiology laboratories .

Objective To analyze the epidemiological characteristics of the imported malaria cases in 20 counties at the bor?der region of Yunnan Province from 2012 to 2014,so as to provide the evidence?based proof for adjusting the strategies in the elimination stage. Methods The malaria epidemic data of the 20 border counties in Yunnan Province from 2012 to 2014 were collected and analyzed by using Microsoft Excel 2010. Results From 2012 to 2014,a total of 1 558 malaria cases were report?ed in the 20 border counties in Yunnan Province,among which,1 336 were imported cases,accounting for 85.75%(1 336/1 558),and 222 were indigenous cases,accounting for 14.25%(222/1 558). The number of the imported cases in the above years took up 80.00%(544/680),89.10%(425/477)and 91.52%(367/401)of the total reported cases in the whole year,re?spectively. Among all the 1 336 imported cases,1 045(78.22%)were infected with Plasmodium vivax,284(21.26%)were in?fected with P. falciparum,3 were infected with P. malariae,3 were mixed infection and 1 was an unclassified case;2 patients died. And 95.58%of the cases were mainly infected in Myanmar(1 277 cases). Young and middle?aged adult of 20-40 years who worked overseas were the predominant(802 cases,60.03%)and most of the cases occurred from April to June of the year (679 cases,50.82%). Those cases mainly distributed in Tengchong(459 cases),Ruili(366 cases),Yingjiang(191 cases)and Mangshi(78 cases). Conclusions The epidemic situation of imported malaria is serious in the border region of Yunnan Prov?ince. Therefore,the surveillance system of malaria control needs to be well planned and managed to ensure timely case detection and prompt response at the elimination and post?elimination stage.