Objective To isolate protoplasts from tube plant leaves of Rhodiola sachalinensis and regenerate plantlets after protoplast culture.Methods Preculture treatment and size of explants,enzyme concentration,mannitol concentration in enzyme mixture related with protoplasts isolation were studied to determine the superior optimized conditions.Results Explants could be used to isolate protoplast without dark preculture,and leaf length should be longer than 1.5 cm.The best incubating enzyme solution contains 1.0% cellulase Onzuka R-10,0.5% Macerozyme R-10,10 mmol/L CaCl2?2H2O,0.1% MES,0.7 mmol/L KH2PO4,and 0.5 mol/L mannitol.The enzyme and explants mixture were shaken for 4 h at 25 ℃.The protoplasts yield and viability were 39.43?106/g fresh weight and 78.6%,respectively.Purified protoplasts were cultured in medium 1/2 MS+1 mg/L 2,4-D+0.5 mg/L ZT+0.5 mol/L mannitol +500 mg/L hydrolysis of casein initially with shallow liquid layers,and calli formed within 40 d.After calli were transfered to MS+1 mg/L 6-BA+0.1 mg/L NAA,adventitious buds were induced from calli.Shoots longer than 2 cm rooted within 30 d when they were transfered to 1/2 MS medium.Conclusion The study provides the scientific base for protoplast fusion in polyploidy breeding of R.sachalinensis.

OBJECTIVE:To survey and objectively evaluate the present situation and trend of the usages of interferon.METHODS:To make a survey of the interferon used in 136 hospitals of 6 cities in the Yangtse valley of China during the period 1998～2000 in respect to the sum of money for drugs,production enterprises etc.and to analyse the situation of clinical use.RESULTS:The sums of money for the interferon were 25 925 795 yuans and 28 512 161 yuans and 28 038 402 yuans in 1998,in 1999 and in 2000,respectively.The proportion of domestic interferon in total consumption of interferon increased year by year.CONCLUSION:With the interferon declining in price and interferon-? having been fixed as a drug in state Basic Drugs for Medical Insurance,its consumption will increase at a high speed.

The present study is designed to establish a xenograft model of human promyelocytic leukemia cell mutant (HL-60-AR) deficient in hypoxanthine guanine phosphoribosyltransferase (HGPRT) in nude mice. A solid leukemia sarcoma developed after subcutaneous inoculation with HL-60-AR cells. Comparative studies of HL-60-AR/Nu tumor cells in nude mice and cultured HL-60-AR cells in vitro revealed virtual identity as shown by light microscopic morphology, ultrastructure of cell, cytochemistry, chromosome analysis, LDH isoenzyme pattern, genetic markers and differentiated characters assay. Up to now, twelve generations haw been transmitted in rive by inoculating with the solid tumor Cells developed in nude mice. This nude mice model in which human leukemia cells grew could be considered as a useful model for in rive studies of human leukemic cells proliferation, differentiation and the screening for anti-leukemia drugs.

Objective To analyze spectral curves and iodine levels in normal adult thyroid by using dual-source CT scanning technique and provide the basis for the differential diagnosis of thyroid space-occupying lesions.Methods A total of 70 subjects were selected for this study,including 38 females and 32 males.They were excluded from thyroid disease by ultrasound examination and had all taken the dual-source CT scan.The image data were transported to the CT workstation,the analysis of iodine levels and spectral curves were completed in theLiver VNCandMonoenergetic modes respectively.The gender differences in ratio iodine content were compared.Results The iodine ratio of thyroid was (1.35±0.19) mg/mlin plain scan,(1.35±0.23) and (1.35±0.21) mg/ml in left lobe and right lobe respectively.There was no statistically significant difference between left and right lobe (P ＞ 0.05),as well as between females and males (P ＞ 0.05).The slopes of the energy spectrum curve in the left and right lobe of the thyroid were 9.98 ± 2.43 and 9.74 ± 1.63,respectively,and 9.88 ± 1.74 and 9.74 ±0.09 for the male and female,respectively.There was no significant difference of slopes of the energy spectrum curve between the left and right lobe,as well as the male and female.The average dose length product of all patients was (208.29 ±21.80) mGy·cm,and the average effective dose was (1.23 ± 0.13) mSv.Conclusions Dual-source dual-energy CT scanning technique have definitive,diagnostic characteristic in the measurement of spectral curves and calculation of the ratio of iodine in normal adult thyroid tissue,and therefore has guidance value in the diagnosis of thyroid space-occupying lesions.

Objective To observe the developmental changes of projection and termination of proprioceptive afferent fibers in the mouse spinal cord. Methods Parvalbumin (PV) immunohistochemistry was used to label the proprioceptive afferents. Single and dual immunofluorescence histochemistry were used to examine the growth pattern of proprioceptive afferents and their relationships with motoneurons in the spinal ventral horn (VH). The stained sections were observed under a confocal laserscanning microscope. Results PV-like immunoreactive (LI) proprioceptive fibers first appeared in the dorsal column on embryonic (E) day 14, then entered the gray matter on El5 and reached the intermediate gray matter and VH more obviously on E16. The number and intensity of PV-LI proprioceptive afferent fibers and punctata increased in the VH with age and reached a maximum during earlier postnatal (P) period (P0-P7). After P14, the number and intensity of proprioceptive afferents gradually decreased. The proprioceptive terminals seemed to form close relationship with motoneurons from E17. Conclusion The present study indicates that the somatotopic organization of proprioceptive afferents in the spinal cord is established during the late embryonic and early postnatal stages. These results provide evidence for understanding the development of the reflex movements.

The present study was designed to examine the developmental changes in projection and termination of nociceptive and proprioceptive afferent fibers in the spinal cord by labeling those two fibers with calcitonion gene-related peptide (CGRP) and parvalbumin (PV)separately in mouse embryos and neonatal pups aged embryonic day 15 to posanatal day 3 (E15 -P3). CGRP-like immunoreactive (LI)nociceptive fibers first appeared in the superficial dorsal horn (DH) at E16. The afferent projections extended laterally to the DH and entered into the deep portions of the DH at E17 and E18. After birth, the projection pattern of CGRP-LI fibers remained unchanged but the intensity of afferent terminals increased in the superficial laminae and their branching patterns became more complicated. In addition,CGRP-LI collaterals that projected into the contralateral DH were also examined after E16. Around birth, the contralateral projections were also found originated from the lateral part of the DH. PV-LI proprioceptive afferents were first observed entering the gray matter at E15 and reached the intermediate gray matter (IG) and the ventral horn (VH) more obviously on E16. The number and intensity of PV-LI fibers increased in the the VH with age and reached a maximum during earlier postnatal period ( P0-P3 ). The proprioceptive terminals seemed to form close relationship with motoneurons in the VH from E17. Our results indicate that the somatotopic organization of nociceptive and proprioceptive afferents in the spinal cord both are established during the late embryonic and early postnatal stages. These results help to understand the development of the sensory transmission in more details.

Objective:To investigate the role of TSLP and NF-κB in the pathogenesis of adenomyosis.Methods:Immunohisto-chemistry was employed to detect the expression of TSLP and NF-κB p65 in the endometrial glandular cell and stromal cell of 35 adeno-myosis patients and 20 hysteromyoma patients as control , and analyze the correlation of the two proteins.The concentration of TSLP was measured by specific ELISA in the serum of healthy subjects besides of the patients with adenomyosis and hysteromyoma .Results:TSLP and NF-κB p65 were both positively expressd in the glandular cells and stromal cells of ectopic and eutopic endometrium of patients with adenomyosis , their expression was significantly higher than that of the control group ( P<0.01 ) , and the expression of TSLP and NF-κB p65 in ectopic endometrium was significantly higher than that in eutopic endometrium ( P<0.01 ).There was positive correlation between TSLP and NF-κB p65 in ectopic endometrium.The concentration of TSLP was higher in the serum of patients with adenomyosis.Conclusion:The TSLP is highly expressed in endometrial glandular and stromal cells of patients with adenomyosis , and there is higher level of TSLP in serum , and there is correlation between the expression of TSLP and NF-κB in ectopic endometrium.

BACKGROUND:Studies have shown that dibutyl phthalate has certain toxicity to the reproductive system of male animals.
OBJECTIVE:To investigate the effects of dibutyl phthalate on secretion of rat ovarian granulose cells. METHODS:Ovarian granulosa cells from 25-day-old female Sprague-Dawley rats were cultured in vitro and exposed to different concentrations of dibutyl phthalate (0, 5, 20, 80μmol/L) for 24 hours. Estradiol and progesterone in ovarian granulosa cells were measured by the method of radio-immunity. Expression of the cytochrome P450aromatase and P450 side chain cleavage enzyme mRNA in granulosa cells were examined by real-time PCR.
RESULTS AND CONCLUSION:The levels of estradiol and progesterone in granulosa cells decreased after exposure to dibutyl phthalate. With the increase of the dibutyl phthalate concentration, the expression of
P450arom and P450scc mRNA decreased gradual y. Dibutyl phthalate could induce reproductive dysfunction and toxicity to female rats.

Objective To investigate the clinical efficacy,safety and the correlation factors of recurrence of radiofrequency catheter ablation in treatment of atrial fibrillation.Methods The clinical data of 127 patients with atrial fibrillation having underwent circumferential pulmonary vein ablation guided by 3-D mapping system were analyzed retrospectively.Then they were followed up for 12 months.The status of patients after ablation and the success rate of ablation was recorded,then the relationship between the factors and the recurrence after ablation of atrial fibrillation was analyzed.Results After a follow up of 12 months,94 patients maintained sinus rhythm,and atrial fibrillation recurred in 33 cases.The first clinical success rate was 74.02％(94/127).Thirty-three patients with recurred atrial fibrillation received ablation again and 11 patients got success.The total clinical success rate was 82.68％ (105/127).The success rate of paroxysmal atrial fibrillation was 88.46％ (92/104) and success rate of persistent atrial fibrillation was 56.52％ (13/23).Univariate analysis showed that the type,the course,the diameters of left atrial,the early recurrence,the body mass index were related to the recurrence of atrial fibrillation,and multivariable Logistic analysis revealed only the type and the diameters of left atrial were independent risk factors responsible for the recurrence of atrial fibrillation (P ＜ 0.05).Conclusion Radiofrequency catheter ablation in treatment of atrial fibrillation is effective and safe,and the type and the diameters of left atrial are independent risk factors responsible for the recurrence of atrial fibrillation.

The fine ultrastructure and localization of acid phosphatase in cell ultrastructuralevel of a HGPRT-human promyelocytic leukemia cell mutant(HL-60-AR)arestudied by scanning electron microscopy,transmission electron microscopy,freezeetching,and electron microscopic cytochemistry techniques.The results of theobservation show that the ultrastructural characteristics of HL-60-AR cells aresimilar to that of HL-60 cells.There are microvilli and ridges over cell surface.Thecells have large nucleus with prominent nucleoli,and numerous nuclear pores.Thereare less developed Golgi complex,expanded rough endoplasmic reticulum andabundant polyribosomes.After treatment with retinoic acid(RA)at 10~(-6) mol/L for 5days,HL-60-AR cells differentiate along myeloid pathway and have a decreasednucleocytoplasmic ratio accompanied with nuclear condensation and segmention.A (?)ignificant increase of specific granules is demonstrated.Microvilli of the cellsdisappear,surface features of the treated cells become more irregular and largeprotrusion and blunt pseudopodia appear.Increase of acid phosphatase content localizedon azurophilic granules(lysosomes)and Golgi complex is showed.The application offour kinds of electron microscopic techniques might provide the best way foridentifying cell ultrastructure.