BACKGROUND: Neural stem cells are always derived from animals, and unsuitable for human transplantation treatment. OBJECTIVE: To explore the in vitro culture methods of human embryonic striatum-derived neural stem cells, and to observe the biological characteristics. METHODS: The human embryonic striatum were separated from the embryo at a gestational age of 8-16 weeks that received induction of labor with water bag, and then the embryonic striatum was in vitro cultured in the serum-free Dulbecco’s modified Eagle’s medium. The cells were passaged after neurospheres formation, and then the cells were induced to differentiation with the Dulbecco’s modified Eagle’s medium/F12 containing 10% fetal bovine serum. RESULTS AND CONCLUSION: The in vitro cultured human embryonic striatum-derived neural stem cells grew rapidly and could express nestin. Colony formation assay showed the cel clone formation rate was 6.0%-7.0%. 5-Bromodeoxyuridine incorporation assay showed the cel proliferation rate was 37.9%. Immunofluorescence staining showed that the cells after induction and differentiation could express Tuj-1, glial fibril ary acidic protein and nestin, but not express myelin basic protein. The results indicate that human embryonic striatum-derived neural stem cells cultured in the serum-free medium can maintain their biological characteristics and have self-renewal capacity, and the cells can differentiate into the neurons and astrocytes induced by the fetal bovine serum.

Adolescent ; Adult ; Female ; Humans ; Humeral Fractures ; complications ; Humerus ; surgery ; Joint Deformities, Acquired ; surgery ; Male ; Orthopedic Fixation Devices ; Osteotomy ; methods

Objective To evaluate treatment results and prognostic factors of 47 patients withprimary urethral transitional cell carcinoma treated with post-operative adjuvant radiotherapy. Methods From October 1998 to October 2008, 47 patients with primary urethral transitional cell carcinoma received postoperative adjuvant radiotherapy. Thirty-one patients had stage T_3/T_4 disease, 7 had lymph node metastasis. Thirty-nine patients had G3 tumor, 13 had stump-positive. The median radiotherapy dose was 60 Gy (36-64 Gy). 81% patients (38/47) were treated with regional irradiation. Results The median follow-up time was 21 months (6 -88 months). The follow-up rate was 92%. The median overall survival time was 35 months (5 -88 months). The 2-and 5-year overall survival rates were 57% and 49%, respectively. In univariate analysis, the median overall survival time was better in patients with stage T_1 or T_2 compared with stage T_3 or T_4 tumor (42 months vs. 19 months,Χ~2 =7. 28,P=0. 007), with age of ≤65 years compared with >65 years (28 mouths vs 18 months,Χ~2 =8.23 ,P =0. 004). There was no significantdifference in the long term survival in patients with non-radical surgery compared with radical mastectomy (21 months vs. 20 months, Χ~2 = 0. 90, P = 0. 344). In multivariate analysis, the stage T_3 or T_4 (Χ~2 = 7. 89, P =0. 005), >65 years old (Χ~2 = 4.85, P = 0. 028), renal pelvis involvement (Χ~2= 5.65, P = 0. 018), and tumor located in the mid or inferior segment (Χ~2=6. 08 ,P =0. 014) were factors associated with poorer prognosis. Conclusions Postoperative adjuvant radiotherapy can improve the efficacy of patients with locally advanced urethral transitional cell carcinoma. Advanced T stage and > 65 years age are associated with poorer prognosis.

Objective To analyze the outcomes of radiotherapy for 74 patients with intracranial ger-minoma. Methods Between 1990 and 2007,74 patients with intracranial germinoma(9 pathologically diag-nosed and 65 clinically diagnosed) were treated with radiotherapy in our hospital. The median age at diagno-sis was 15 (range 5-45) years. Radiation treatment fields varied among patients, including craniospinal irra-diation(CSI), whole brain irradiation, whole ventricular irradiation with primary tumor boost, and involved-field irradiation only to the primary tumor plus margin. The dose was 38.5 -50.0 Gy to the tumor,18-25 Gy to the whole brain/ventricular,and 21-25 Gy to the whole spinal cord in fractions of 1.6-2.0 Gy per day,5 fractions per week. Results The median follow-up time was 80(range 12-168) months and the fol-low-up rate was 97%. Fourteen patients had been followed up for over 10 years. The 1-,5- and 10-year o-verall survival rates were 99% ,96% and 93%. The corresponding disease free survival rates were 97%, 90% and 83%, respectively. Relapses occurred in 9 patients. For the 6 patients with in-field relapse, the dose to the tumor was 38.5-40.0 Gy in 3 patients, 41-45 Gy in 2 and 46-50 Gy in 1. Relapse in the spinal cord was found in 3 patients and none of them received spinal irradiation. Twenty-one patients re-quired hormonal replacement therapy because of radiation induced hypofunction of prehypophysis. Conclu-sions Radiotherapy alone is a curative treatment for intracranial germinoma. The proper dose should be de-termined by tumor numbers and the examination of cerebrospinal fluid.

Objective To study the efficacy,late complications and prognostic factors of postopera-tive radiotherapy for bladder cancer: Methods Between July 1995 and January 21307,109 patients with primary bladder cancer who had received adjuvant pelvic radiation therapy were retrospectively analyzed. The age ranged from 41 to 94 (median 68) years. There were 29 patients with T1 disease,60 with T2,17 with T3, 3 with T4, and 7 with positive lymph node. Three-dimensional radiotherapy was given with a median total dose of 49.2 Gy(39.3 -62.2 Gy) in conventional fractionation after surgery. Field-in-field intensity modu-lated radiotherapy was given to 57 patients. Results The total follow-up rate was 98% ,with a median fol-low up time of 36(2-144) months. The 1-,3- and 5-year local control rate was 63% ,47% and 42% ,respec-tively. The 1-,3- and 5-year overall survival rate was 80% ,48% and 37% ,respectively. Among the 109 pa-tients,33 died of tumor progression or metastasis,42 showed local recurrence,and 22 had lymph node metas-tasis. According to the RTOG criteria,grade 1,2,3 and 4 radiation related early urological side effects were 22% ,59% ,12% and 1% ,respectively;The corresponding late urological side effects were 29% ,28% ,2% and 1%, respectively. Two patients underwent whole bladder cystectomy due to the radiation related side effects. Conclusions Postoperative three-dimensional radiotherapy, achieving high response rate with tol-erable side effects, can be offered as an alternative option to the radical cystectomy in the bladder conserving treatment.

Objective To analyze the efficacy,late complications and prognostic factors of post-op-erative radiotherapy for cervical cancer. Methods From Nov. 1999 to Feb. 2005,114 patients with cervi-cal cancer received adjuvant pelvic radiotherapy after radical hysterectomy and pelvic lymphadenectomy. The median age was 42.5 (24 to 72) years old. According to the FIGO staging system,6,51,18,26 and 13 pa-tients had stage ⅠA, Ⅰb1, Ⅰb2, ⅡA and ⅡBdisease. The pathological type was squamous cell carcinoma,ade-nocarcinoma, squamous-adenocarcinoma and undifferentiated carcinoma in 92,19,2 and 1 patients, respec-tively. The whole-pelvic external beam irradiation of 50 Gy (40 to 60 Gy) was given with 6 MV or 15 MV X-ray beams using four-field box technique. Eighty-one patients received intravaginal brachytherapy of 16 Gy (4 -30 Gy in 1 -6 fractions) 4 weeks after the beginning of radiotherapy, with the referrence point being at 0.5 cm under the vaginal mucosa. Eighty-seven patients received preoperative and/or concurrent chemother-apy. The survival and independent prognostic factors were analyzed by Log-rank method and Cox model. Results The median follow-up time was 26.0 (5 - 75) months. The overall survival rate, disease-free sur-rival rate and local control rate were 93.1%, 88.1% and 94.6% at 2-year, and 75.7%, 62.3% and 85.6% at 5-year,respectively. The independent prognostic factors were lymph node metastasis and positive surgical margin for overall survival, positive surgical margin for local control, and stage, uterine body invasion and positive surgical margin for disease-free survival. Sixteen patients ( 14% ) had distant metastasis, and the most common sites were the lung,inguinal region,bone,liver and brain. According to RTOG grading sys-tem, the incidence of late gastrointestinal side effects of grade 1,2 and 3 was 11.4%, 11.4% and 3.5%. The corresponding genitourinary side effects were 14.0% ,6.1% and 0.9%, respectively. The incidence of leg lymphedema was 7 % . Conclusions Post - operative radiotherapy can achieve good local control in cervical cancer with acceptable late side effects. Distant metastasis is the main cause of death.

Carcinoma ; genetics ; metabolism ; pathology ; surgery ; China ; Humans ; Male ; Oncogene Proteins, Fusion ; genetics ; Prostatic Hyperplasia ; genetics ; metabolism ; pathology ; surgery ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-ets ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serine Endopeptidases ; genetics ; metabolism

OBJECTIVETo establish a simple but effective method of laser scanning confocal microscopic imaging for Ca2+ oscillations of pancreatic acinar cells in adult mice.

METHODSPancreatic acinar cells from adult Kunming mice were isolated acutely with collagenase, and then loaded with fluo-4-AM, a Ca2+ indicator. A laser scanning confocal microscope armed with 488 nm laser was employed to record the dynamic fluorescent signals in-time and synchronously while acetylcholine (ACh) was added in the pancreatic acinar cells.

RESULTS(1) The classic pancreatic acinar cell Ca2+ oscillations were induced by a certain concentration of ACh (100 nmol/L) successfully and steadily, which could be blocked by atropine completely. (2) Plasmic Ca2+ oscillations from different parts of one acinar cell were usually with different amplitudes and almost the same frequencies. But both of amplitudes and frequencies were different among different cells. (3) The acinar cell Ca2+ oscillations were induced by ACh in a concentration-dependent manner.

CONCLUSIONThe laser scanning confocal microscopic imaging for adult mouse pancreatic acinar cell Ca2+ oscillations was established successfully. The features of being easy to use, direct to see lively, high efficiency and good flexibility make it a popular tool for researchers to choose.

Acinar Cells ; chemistry ; Animals ; Calcium ; analysis ; Calcium Signaling ; Cells, Cultured ; Mice ; Microscopy, Confocal ; methods ; Pancreas ; cytology

Zhigancao Decoction is from Shang Han Lun,which is widely used to treat cardiovascular diseases nowadays,such as arrhythmia,myocarditis,heart failure and so on.Moreover,according to records in Wai Tai Mi Yao:Zhigancao Decoction can also be used to treat lung"atrophy". This article introduced application of Zhigancao Decoction in treating respiratory diseases with good efficacy, which could provide new ideas for the TCM treatment of advanced respiratory diseases.

Objective To investigate the effects of anxiety and depression on the quality of life (QOL) in patients with irritable bowel syndrome with diarrhea (IBS‐D) and the difference in gender .Methods IBS‐D patients met the Rome Ⅲ diagnostic and subtyping criteria were consecutively enrolled .The intestinal symptoms , psychological status , and QOL of patients were evaluated using IBS‐specific symptom questionnaires , the Hamilton anxiety scale (HAMA) ,the Hamilton depression scale (HAMD) ,and the Chinese Version of IBS‐QOL instrument .The data were analyzed by chi‐square test variance analysis ,t test or Spearman rank correlation analysis .Results A total of 155 IBS‐D patients were enrolled .Among them ,115 were complicated with anxiety and/or depression .The number of male and female with comorbid psychological disorders was 69 cases(71 .13% ) and 46 cases(79 .31% ) ,respectively ,and the difference was not statistically significant (χ2 = 1 .267 ,P= 0 .26) . Compared with 40 patients without anxiety or depression ,there was no statistically significant difference (F=1 .143 ,P> 0 .05) in the scores of intestinal symptoms in 59 patients with comorbid anxiety alone and 56 patients with anxiety and/or depression (20 .85 ± 5 .84 vs .21 .71 ± 7 .47 vs .22 .87 ± 6 .09) .The total IBS‐QOL score of the 155 patients was 71 .61 ± 19 .22 .There was negative correlation between IBS‐QOL score and HAMA , HAMD scores(r= -0 .262 and -0 .268 ,both P= 0 .001) .The total IBS‐QOL score of patients with comorbid anxiety and depression or depression were lower than that of patients without anxiety or depression (66 .05 ± 22 .88 vs 77 .22 ± 15 .35 , F = 4 .412 , P = 0 .005) .Except health worrying and sexual ,the scores in six dimensions(including dysphoria ,interference with activity ,body image ,food avoidance and social reaction) were all significantly decreased (F= 3 .309 ,3 .279 ,4 .177 ,3 .765 ,6 .041 and 3 .830 , P= 0 .013 ,0 .012 ,0 .013 , 0 .007 ,0 .001 and 0 .010) .In male patients ,the total score of IBS‐QOL and the scores of dysphoria ,body image and social reaction of patients with anxiety and depression or depression were lower than those of patients without anxiety and depression (t = 2 .143 ,2 .110 ,2 .279 ,3 .061 ; P = 0 .036 ,0 .039 ,0 .027 ,0 .003) .In female patients ,the score of food avoidance domain significantly decreased (t= 2 .812 ,P= 0 .008) .The scores of IBS‐QOL and six dimensions (including dysphoria ,interference with activity ,body image ,food avoidance and social reaction) of patients with history of severe mental disorders and being abused were even lower than those patients without such histories (t= 3 .241 ,3 .433 ,2 .499 ,2 .296 ,2 .514 ,3 .413 and 2 .601 ;P= 0 .001 ,0 .001 ,0 .014 , 0 .023 ,0 .013 ,0 .001 and 0 .010) .Conclusions The QOL of patients with IBS‐D was significantly affected by psychological factors with gender differences .The approaches to improve the psychological status of IBS patients were helpful to enhance the efficacy of comprehensive therapy .