Object To establish a calli culture system for the rapid propagation of Speiraea japonica L. f.. Methods Callus and shoot induction were carried out on MS, 6, 7 V or B 5 media with different parts of the plant such as stem tip, tender leaves and petiole as explants. Results A calli culture system was established for the rapid propagation of S. japonica. Conclusion MS cultural medium was found to be most suitable for calli induction. MS with 2.0 mg/L 2,4 D+0.3 mg/L KT can induce calli when the explants were used for the induction, with stem tips being the most satisfactory explant. Clusters of seedlings can be induced on MS+2 mg/L BA+0.1 mg/L NAA and when these seedlings were transferred to 1/2 MS +0.25 mg/L BA+0.5 mg/L NAA medium, root were developed to give young seedlings.

BACKGROUND:There is few mesenchymal stem cells(MSCs) in the bone marrow,about one BMSC in 1?104-1?105 monocytes.Following in vitro isolation,purification and amplification,it is satisfactory for in vivo requirement.OBJECTIVE:To observe biological characteristics and potentiality of multi-directional differentiation of BMSCs in vitro.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Institute of Urinary Surgery,Union Hospital Affiliated to Fujian Medical University from October 2007 to December 2008.MATERIALS:A total of 30 clean male Sprague Dawley rats of inbreeding line were supplied by Silaike,Shanghai,China.METHODS:BMSCs from rats were isolated and cultured in vitro by using the whole bone marrow adherence method.When 80%-90% confluency,BMSCs were digested by trypsin.At the third passage,BMSCs were induced to differentiate into osteoblasts and adipocytes.MAIN OUTCOME MEASURES:Cell morphology was observed under the inverted phase contrast microscope.BMSCs surface markers were detected using flow cytometry.Osteogenic ability was examined by alizarin red staining.Adipogenic ability was measured by Oil red O staining.RESULTS:The primary cells and the passage cells were mostly fusiform in shape,to be similar to fibroblasts.Cell still kept a high potential of growth and amplification following over 10 subcultures.At the third passage,BMSCs were positive for CD44,CD90,CD106,but negative for CD34,CD45,CD11b.Following 21 days of osteogenic induction,cell alizarin red staining showed that alizarin red was positive in osteoblasts.Following 2 weeks of adipogenic induction,oil red O staining showed that red lipid droplet existed in adipocytes.CONCLUSION:Whole bone marrow adherence method can isolate,purify and amplify BMSCs in vitro.The obtained cells have general biological characteristics of MSCs,and also have potentiality of multi-directional differentiation.

Objective To evaluate the efficacy of microvasoepididymostomy for the treatment of obstructive azoospermia. Methods Ninety-eight patients were confirmed as obstructive azoospermia by infertility investigations.The mean (range) age was 31 (20-43) years.The mean duration of obstruetion was 4 years.All patients were seen by the surgeon for a complete history and physical examination.Semen analyses proved azoopermia,serum levels of sexual hormone were normal and testicular function for production of sperms were normal certified by biopsy.All patients with suspected epididymal or vasal obstruction were offered scrotal exploration.The decision for microvasoepididymostomy was made during surgery,based on the pateney of the vas towards the abdomen documented by saline and sperm fragments containing in the epididymal fluid on intraoperative light-microscopic examination.Microvasoepididymostomy was performed in 58 patients documented epididymal obstruction The initial semen analysis was then done after usually 3 months postoperatively.Patency was arbitrarily defined as＞10 000 sperm/mL in ejaculate in at least one semen analysis after surgery.Postoperative patency rate and postoperative impregnation rate were followed. Results Fifty patients were followed up for 3-29 months,8 cases lost.Sperm was found by semen analysis in 36 patients.Sperm density was 4×104-2×108 sperms/ml and motility was 2%-70%.Semen analysis reveals azoospermia in 6 patients and the patients were counseled to undergo further testing to determine the ultimate outcome of the procedure.Natural conception occurred in 14 patients followed for more than 12 months.The overall pateney rate was 72%(36/50).Among patients with a follow-up of＞6 months,the natural paternity rate was 28%(14/50).The median time to achieve a natural pregnancy was 6.6 (4.0-10.0)months. Conclusion A more favourable patency can be achieved using microsurgical intussusception vasoepididymostomy and a part of patients with obstructive azoospermia can be cured by this means.

Objective To compare the circumferential approach and suprapubic approach for correcting the concealed penis. Methods Thirty-four patients received circumferential approach (group A,18 cases)or suprapubic approach(group B,15 cases)randomly for the concealed penis.The length of penis without erection before and after surgery treatment,patients'satisfaction and surgical complications were evaluated.All the patients were followed at least 6 months after operation.Results The penile length before and after surgery in group A were(1.78±0.41)cm vs(3.97±0.47)cm,the length of the postoperative penis was significantly longer than that of the preoperative penis(P<0.01).83%of those in group A(15 cases)were satisfied with the cosmic results.The length of preoperative and postoperative penis in group B were(2.07±0.53)cm vs(4.05±0.81)cm respectively,the length of the postoperative penis was significantly longer(P<0.01).Satisfaction was 87%(13 cases).The postoperative penile length between the 2 groups was not different(P> 0.05).Prepuce edema was reported to Occur in 56%and 13%in group A and group B and spontaneously disappeared within 3 months.Fat synchysis occurred in 2 of group B. Conclusions The cosmic effect of the 2 surgical approaches is similar,but circumferential approach is simpler.There is no serious complication in those receiving circumferential approach.

Objective To investigate the diagnostic value of the T-SPOT.TB in connective tissue disease(CTD) combined with tuberculosis.Methods This is a case-control study.Forty-four patients with CTD combined with tuberculosis were enrolled from Xiangya Hospital of Central South University from September 2011 to July 2012.Another forty-four CTD patients without tuberculosis were evaluated as a control group.The diagnostic value of T-SPOT.TB and risk factors of the false negative results by T-SPOT.TB were analyzed.Results The sensitivity of T-SPOT.TB (70.5％,31/44) was significantly higher than that of TST(27.3％,12/44) for CTD combined with tuberculosis patients (x2 =16.42,P ＜ 0.001).The specificity of T-SPOT.TB and TST were 93.2％ (41/44) and 88.6％ (39/44),respectively.There was no significant difference between the specificity (x2 =0.14,P =0.711).The positive predictive value of T-SPOT.TB was 91.2％ (31/34).The negative predictive value was 75.9％ (41/54).Youden's index was 0.64,and the positive likelihood ratio was 10.3.All the index were higher than that of TST (0.16 and 2.4).While the negative likelihood ratio which was 0.32 was lower than that of TST (0.82).When spot forming cell frequencies of T-SPOT.TB of PBMC was set to 38SFCs/106 PBMC,it had the best cut-off value.Age,use of glucocorticoids or immunosuppressant therapy,lymphocytopenia and hypoalbuminemia were not associated with false negative T-SPOT.TB assay.Conclusion The T-SPOT.TB assay is much more useful than TST for diagnosing CTD combined with tuberculosis.

To study the chemical constituents of Lysimachia patungensis Hand.-Mazz., silica gel column chromatography, reverse phase ODS column chromatography, MCI and Sephadex LH-20, were used to separate the 95% EtOH extract of the whole plant of Lysimachia patungensis Hand.-Mazz.. The structures of the isolated compounds have been established on the basis of chemical and NMR spectroscopic evidence as well as ESI-MS in some cases. Twelve phenolic compounds were obtained and identified as quercetin-3, 3'-di- O-alpha-L-rhamnoside (1), myricetrin (2), quercitrin (3), rutin (4), 2-hydroxynaringenin-4'-O-glucopyranoside (5), naringenin 7-O-glucopyranoside (6), liquiritin apioside (7), licochalcone B (8), tetrahydroxymethoxy chalcone (9), methyl-p-coumarate (10), 2, 4, 6-trihydroxy acetophenone-2-O-glucopyranoside (11) and vaccihein A (12). Among them, compound 1 is a new compound, and compounds 5, 11 and 12 are isolated from the genus Lysimachia L. for the first time, and the others are isolated from the plant for the first time.

Surgical excision and postoperative radiotherapy are widely accepted therapeutic modalities for sinonasal teratocarcinosarcoma (SNTCS). Previous studies have shown that intensity-modulated radiation therapy (IMRT) is safe and effective for the treatment of some sinonasal malignancies. We hypothesize that use of IMRT for SNTCS may result in clinical benefits. We report here two cases of SNTCS that were treated by IMRT. One patient was given extensive IMRT involving elective neck irradiation. Follow-up examinations revealed no recurrence and metastasis 3.5 years after IMRT. Another patient simultaneously suffered from multiple systematical diseases and was administered involved-field radiotherapy. He was found to have a marginal recurrence during the follow up and eventually died 8 months after disease diagnosis. It was suggested in this study that appropriate use of IMRT for the treatment of SNTCS may achieve excellent local control.

This study mainly explored the pulse parameters in children with RRTI with different TCM syndromes,aiming at providing therapeutic indexes and objective basis for its diagnosis and treatment.Three hundred and forty-eight cases of RRTI were divided into five groups,including the group of qi deficiency in the lung (or Fei Qi Xu,FQX),the group of invasion of the lung by wind-heat (or Feng Re Fan Fei,FRFF),the group of invasion of lung by wind-cold (or Feng Han Fan Fei,FHFF),the group of obstruction of phlegm-damp in the lung (or Tan Shi Zu Fei,TSZF) and the group of obstruction of phlegm-heat in the lung (or Tan Re Yong Fei,TRYF).65 children of good health were involved in the control group.Z-BOX pulsemeter apparatus was applied to the paraticipants for analyzing their pulse parameters.As a result,it was found that values of h1,h3,h4,h5,t and h4/h1 of RRTI children decreased,compared with the children of good health (P ＜ 0.01);while h1,h3,h4 and h5 of children in FQX group declined (P ＜ 0.01);and the values of h4,h5,t,w,h3/h1,h4/h1 and h5/h1 of children in FRFF group went down (P ＜ 0.01);while the values of h1,h3,h4,h5,t,w,h3/h1 and h4/h1 of TRYF group fell (P ＜ 0.01);and those of h5,t and h5/h1 of children in FHFF group decreased (P ＜ 0.01).Compared with FQX group,h1 value of FRFF group increased (P ＜ 0.01),while the values of w,h3/h1,hs/h1 and w/t of FRFF group declined (P ＜ 0.01);and the h1 value of TSZF group boosted (P ＜ 0.01),while the value of w and h3/h1 of TRYF group decreased (P ＜ 0.01);and the h5/h1 value of FHFF group fell (P ＜ 0.01).In comparison with FRFF group,the values of t,w and h5/h1 of TSZF group went up (P ＜ 0.01),while the values of h1 and h3 of TRYF group declined (P ＜ 0.01).In comparison with TSZF group,the values of h3,h4,t and w of TRYF group went down (P ＜ 0.01),and the t value of FHFF group decreased (P ＜ 0.01).In conclusion,the pulse parameters of RRTI children can be recognized as objective indicators for TCM syndrome differentiations.

Objective To investigate the expressions of fatty acid-binding protein 5 (FABP5)and dihydroli-poamide dehydrogenase(DLD)in skin lesions of symmetrical acrokeratoderma(SAK), and to explore their significance. Methods Biopsy specimens were obtained from skin lesions on the wrists and perilesional skin of 9 patients with SAK, and from normal skin in the wrists of 9 healthy volunteers (control group). Reverse transcription PCR (RT-PCR)and immunohistochemical staining were performed to measure the expressions of FABP5 and DLD in these specimens. Results RT-PCR showed no significant differences in the mRNA expressions of FABP5 or DLD between lesional, perilesional and normal control skin specimens(both P > 0.05). Immunohistochemically, there was a significant increase in the extent and intensity of staining for FABP5 in SAK lesions. Concretely speaking, FABP5 was strongly expressed in the stratum corneum, granular and spinous layers in SAK lesions, but weakly expressed in the stratum corneum, granular and spinous layers in perilesional skin, and only in spinous and basal layers in normal control skin. The expression of DLD decreased in SAK lesions, and was observed only in the stratum corneum and spinous layer in a few cases of SAK. However, the full-thickness epidermis stained positive for DLD in perilesional skin, with the nuclei and cytoplasm both stained deep brown. Conclusion The overexpression of FABP5 in SAK lesions may participate in dysdifferentiation of keratinocytes, while the down-regulation of DLD expression suggests an imbalance in energy metabolism.

Objective To discuss the pathogenesis of the statin-induced rhabdomyolysis in renal transplant recipients.Methods We presented two renal transplant recipients who developed rhabdomyolysis in 2012 in our hospital.The clinical presentation,laboratory results,diagnosis and treatment of the two patients were analyzed retrospectively.The basic immunosuppressive agent of two patients was cyclosporine A.The recipients developed rhabdomyolysis following simvastatin lipidlowering therapy,and one patient suffered acute renal failure simultaneously.Acute tubular injury was confirmed by renal biopsy.Finally,the symptoms of the two patients were relieved completely,creatine kinase (CK) returned to normal after the satins discontinued and saline,sodium bicarbonate and diuretics were given.The renal failure patient underwent plasma exchange and CRRT,and the renal function returned to normal.Results The level of cyclosporine A should be monitored when the renal transplant patient was given statins,especially whose basic immunosuppressive agent was cyclosporine A.At the same time we should pay more attention to the symptoms of the myotoxic side effects and avoid using the drug which was also metabolized by CYP3A4.Conclusion Physicians should be aware of the potential risks of combined therapy of statins which are metabolized by P450CYP3A4 and cyclosporine A in transplant patients.If using it is advisable to begin with small dosage and monitor the CK level.