Objective To develop a method for determination of activity and viability of Hansenula polymorpha by dual fluorescent staining with carboxyfluorescein diacetate(CFDA)and propidium iodide(PI).Methods The time durations(5,10,20,40,60 and 120 min for CFDA,5,10,15,20 and 25 min for PI)and working solution concentrations(50,100,200and 300 μg for CFDA,2,10,20 and 30 μmol/L for PI)for the dual fluorescent staining were optimized by single factor test. Under the optimal condition,the H.polymorpha samples at theoretical survival rates of 0,25%,50%,75% and 100%were determined,of which the fluorescent intensity was observed under fluorescent microscope,and the gray value was analyzed by ImageJ software. The live and dead cells were counted,based on which the actual survival and death rates were calculated. Meanwhile,the relationships of actual gray value,actual survival rate and actual death rate to the corresponding theoretical values were analyzed. Activity and viability of three batches of cultured H.polymorpha were detected by CFDA-PI dual fluorescence staining.Results The optimal time durations for staining with CFDA and PI were 60 and 5 min,while the optimal working solution concentrations were 200 and 2 μmol/L,respectively. The actual gray value,actual survival rate and actual death rate of H.polymorpha samples at various theoretical survival rates were significantly correlated to the corresponding theoretical values(R~2=0. 998 3～0. 999 2,P < 0. 05). The CVs of activity and viability values in three detections of three batches of H.polymorpha culture were 3. 20%～4. 03% and 1. 10%～2. 27%, respectively.Conclusion The CFDA-PI dual fluorescent staining was successfully developed,which may be used for determination of activity and vitality of H.polymorpha.

Asian Continental Ancestry Group ; Humans ; Language ; Medicine, Chinese Traditional ; Translations

As a kind of inherent membrane surface molecules of our human body,connexins are closely related to the cell growth,proliferation and differentiation,which are highly expressed in kidney.Many kidney diseases have a close relationship with connexins' altered expression and disordered function.This paper is dedicated to review the characteristics of connexins molecules,the expression and corresponding functions in the kidney tissues,as well as the possible regulation mechanisms in the kidney tissues in order to explore their applications and value in medical study.

OBJECTIVE: In China, there are little literatures about nonspecific low back pain(NLBP) and no standardized diagnostic classification of it. This paper reviews the clinical and basic studies on NLBP, and discusses the clinical features and progress in related studies. It is necessary for rehabilitation and intervention of NLBP science norm.DATA SOURCES: Medline, China Wanfang, China Biomedicine (CBM),China Academy Journal(CAJ) and its VIP database.STUDY SELECTION: By searching "nonspecific" and"low back pain" for all data and information about NLBP, we obtained 163 English articles and 22 Chinese ones. These documents were further restricted by "in title" and "2000-2004" to obtain subject documents in the latest 5 years(28 English and 9 Chinese documents) . The main references of important documents were manually searched for full-text articles. Types of these documents were basic study, clinical study.DATA EXTRACTION: All the related information about NLBP in the full-text documents was classified and extracted.DATA SYNTHESIS:Low back pain is generally classified into 3 types according to the present clinical guidelines: NLBP, specific low back pain and sciatica. NLBP constitutes 85% of low back pain and its cause is not clear although it is related to many risk factors. Of the treatment methods for NLBP, all are selective ones.CONCLUSION: Standard classification of diagnosis of low back pain needs to be established to guide clinical practice. Prevention is better than treatment. Tertiary prevention combined with multidisciplines is a promising way to relieve pain and prevent recurrence and disability.

Objective:To summarize the experiences of the diagnosis and surgical treament for thymoma.Methods:Sixty-five patients with thymoma,surgically treated in our department in the period 1970 to 2000,were retrospectively analysed in respect to the surgical approach,simple thymoma resection and extended thymoma resection.Results:Imaging examination,such as chest X-ray,was the main diagnostic method.Myasthemia gravis was common concomitant disease with an incidence of 43.1%(28/65).Extended thymoma resection was superior to simple resection in therapeutic efficacy.Radiotherapy and chemotherapy could raise therapeutic effects in tumors over stage-Ⅱ.Conclusion:It is inappropriate to adopt simple thymoma resection.No matter which stage the tumor is,extended resection is the effective method to decrease the recurrent rate.

Used three different methods(ELISA、EFIA、Tr-FIA)to measurate concentration of thyroid stimulating hormone(TSH)in dried blood samples on filterpaper of 32 Uygur neonates again,that were collected two or three year ago.At the same time,collected dried blood samples on filterpaper from the 32 children.Measurated concentration of TSH in the dried blood samples and did clinical examination.The results revealed that there is definite error in different methods.Eliminating measure error,the cut off of mensurating congenital hypothyroidism with dried blood samples on filterpaper are as follows:It is negative,if the concentration of TSH in dried blood samples on filterpaper is≤10?U/ml.It is dubious,if the concentration of TSH in dried blood samples on filterpaper is 10~20?U/ml.These neonates will be counterchecked in definite time to diagnose.It is positive,if the concentration of TSH in dried blood samples on filterpaper is≥20?U/ml.

0.05). Conclusions:With a low rate of missed diagnosis which had no relation with the degree of CIN, colposcopic cervical biopsies was a convenient, economical,less wounded and a reliable method in diagnosing cervical intraepithelial neoplasia.

Objective:To establish a high performance liquid chromatography/tandem mass spectrometry(HPLC-MS) method for determination of cyclosporin A level in the whole blood.Methods: A Hypersil C_(8) column(65 ℃,4.6 mm ?150 mm,5 ?m) was used with a mobile phase of acetonitrile-acetic acid(pH 4.50) 7030 at a flow rate of 1.0 ml?min~(-1).Atomospheric pressure electronic spray ionization was used to quantify cyclosporin A,with the capillary voltage being 4 000 V,the flow of drying gas being 8.0 L?min~(-1),nebulizer pressure being 30 psi,and fragment voltage being 140 V.Results: The calibration curve of cyclosporin A was linear in a range of 26.68-1 067 ?g?L~(-1): y=0.001 1x-0.004 6(r=0.999 3).The lowest limit of detection was 4.35 ?g?L~(-1).The method recoveries of low,medium and high concentrations cyclosporin A were((89.81?4.91)%,)(104.5?3.73)%,and(95.17?2.33)%,respectively;the extraction recoveries of 3 concentrations were(77.64?4.38)%,(78.01?3.79)%,and(83.02?1.59)%,respectively.The intra-day and inter-day RSD was less than 6%,and the RSD was less than 5% in 150 days.Conclusion: The present method is fast,sensitive and selective.It is suitable for clinical monitoring of cycolsporin A.

1 The history and main problems in prenatal diagnosis and clinical diagnosis of inherited diseasesPrenatal diagnosis is to diagnose the fetal congenital defects and inherited diseases. In the earlier 1970s, thegroup of prenatal diagnosis in Peking Union Hospital was founded by Dr. Qiao-zhi LIN?Meanwhile, under the lead-ership of Dr. LIN, the group successfully completed the culture of amniotic fluid cells and began to diagnose fetalchromosomal diseases in the second trimester of pregnancy. In 1988, they succeeded in diagnosing chromosomaldiseases by taking the chorionic villus in the early pregnancy. Since then, the genetic and prenatal counseling tothose women who previously had children with severe defects were gradually developed. The work of the Peking U-nion Hospital developed prenatal diagnosis in our country. In 1990s, due to the unsettled problems about the posi-tion of technicians in prenatal and genetic disease diagnosis in medical institutions, the falling behind of standardtraining, the lost of person with proper ability, and the lack of standard criteria for some highly difficult techniquessuch as culturing amniotic fluid cell,the chorionic villus sampling,clinical diagnosis of chromosomal and geneticdiseases, medical friction appeared in some regions.At the same time, this type of clinical service, with difficulttechnique and high risk, is short of lawprotection, which reduced the enthusiasm of advanced clinical service. Inthe whole country, there is a shortage of persons with ability and clinical mature techniques?The Standardized crite-ria universally used in developed countries were not introduced to our country in time. The above factors leaded tothe lag-off on prenatal and genetic disease diagnosis in clinical service.

To provide new thinking on improving the teaching quality of health statistics of medical postgraduates,the author advanced his thoughts and opinions on teaching reform of health statistics combined with medical research methods.WTHZ〗