Objective To test whether feeder cells derived from mouse embryonic stem cells(mESCs) could support the growth of mESCs themselves. Methods mESCs were induced to form mouse embryoid bodies(EB),and then fibroblast-like cells were derived from further differentiated mEB(mEB-dF),which served as feeder cells.The undifferentiation of mESCs grown on mEB-dF was confirmed by morphological analysis,colony efficiency and cell differentiation rate of mESCs,immunocytochemistry,alkaline phosphatase staining and RT-PCR.The pluripotency of mESCs grown on mEB-dF was examined by RT-PCR,inducing their differentiation in vivo and in vitro. Results(Forty-eight) fibroblast-like cells lines were derived from the same EB at three periods(d 10,d 15 and d 20),and five of them,mostly derived from d15 EB,were able to maintain mESCs in undifferentiated status and pluripotential ability over 10 passages.mESCs cultured on these feeder cell lines expressed alkaline phosphatase and specific mESCs markers,including SSEA-1,OCT-4,NANOG,and formed EB in vitro and teratomas in vivo.However,the majority of mEB-dF lines(43/48) has no such ability. Conclusion This study not only provides a novel feeder system for mESCs culture,avoiding lot of disadvantages of mouse embryo fibroblasts used as the feeder,but also indicates that fibroblast-like cells derived from mESCs take on different functions.The molecular mechanism of different function of these fibroblast cells is worthy of further investigations.

BACKGROUND:Gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite materials were developed in our previous studies. OBJECTIVE:To observe the capability of gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite materials in the repair of rabbit radius defects. METHODS: Thirty healthy adult New Zealand rabbits were selected to make critical-size lacunar bone defects of the upper radius (15 mm×6 mm). Then, the rabbit models were randomized into experimental group (n=15), autogenous bone graft group (n=10) and blank group (n=5). Gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite materials were implanted into radial bone defects in the experimental group. Bone defect in blank group was implanted without any materials; in the autogenous bone graft group, the contralateral radius with same length was taken and implanted into the defect. General observation, histological observation and X-ray observation were performed respectively at 2, 4, 8, 12 weeks postoperatively. RESULTS AND CONCLUSION:At 12 weeks after operation, the experimental group showed obvious new blood vessels at the defect region, complete bony union and disappearance of the composite implant, but lamelar bone structure appeared, smal blood vessels were visible, the edge of new bone was connected to the original bone edge, exhibiting a continuity of bone, the bone density was slightly lowered, and the defect region became unobvious. In the autogenous bone graft group, bony union and trabecular bone reconstruction were distinct, the lamelar bone became mature, the medulary cavity was recanalized, the fracture line disappeared completely, and the bone density was completely consistent with that of the original bone. In the blank group, there was no obvious bone formation, which led to bone nonunion, and there were a great amount of fiber tissues and inflammatory cel infiltrated. To sum up, the gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite material can obviously promote the repair of critical-size bone defects, and the repairing effect is basicaly the same with that of autologous bone grafting.

ObjectiveTo observe the effect of sport walking on heart and lungs functions in old women.MethodsForty-two old women aged from 60 to 69 had sport walking for 4 months; the speed and intensity were controlled according to their heart rate respectively. The indexes of respiration, circulation and rheoencephalography were measured before and after exercise.ResultsAfter exercise, blood-pumping function of heart improved obviously, stroke volume increased from (65.22±11.41)ml to (72.10± 10.78)ml, ejection fraction increased from (60.10±5.03)% to (68.78±6.25)%, while heart rate declined from (77.45±8.69) times/min to (7.89±8.21)times/min, capacity increased from (2.86±0.36)L to (3.34±0.53)L, and maximal voluntary ventilation for every minute increased from (96.14± 15.21)L to (114.02±16.01)L, significantly different compared with that before treatment ( P<0.01). The fluid time of rheoencephalography reduced from (0.171±0.058)s to (0.128±0.049)s ( P<0.01).ConclusionSport walking under proper intensity can improve the function of respiration and circulation system in old women, so it is a good way to keep health for the elders.

Objective To explore the clinical features,diagnosi s and prognosis of incontinentia pigmenti.Methods Analyzing and summarizing the clinical characteristic, diagnosis and prognosis of neonatal incontinentia pigmenti in 6 neonatal infants that were hospita- lized in our department during the period from January 1 998 to December 2003 were studied,and some relevant literature were reviewed. Results 1.Three of 6 infants were male which was unusual;2.Four infants had typical skin lesions at birth and 1 case at 6 days old.Four cases had typical 3 stages o f skin lesions including the erythematous and vesicular inflammatory stage,verr ucous lesions and hyperkeratosis stage,macular hyperpigmentation stage,but the re was overlap;3.Four infants were complicated by central nervous system involv ement (two cases presented mental retardation,2 infants were temporary damage). Two cases were complicated by ocular manifestations ( one case had optical nerve atrophy and blind in left eye,the other had severe bilateral retinal lesions); 4.On specific examination 5 infants were diagnosed by skin biopsy.Gene analysis was made in 1 case,but we didn′t find the mutations of NEMO. Conclusions Incontinentia pigmenti is a rare X-linked dominant multisystem disease.It may be misdiagnosed in the initial stages.Except typical clinical features,skin biops y and gene analysis are main evidence for diagnosis.Early detection and interven tion are important for prognosis. J Appl Clin Pediatr,2005,20(2):123-125

Objective To summarize the clinical experiences of mini-incision urological surgery.Methods The clinical data of 43 patients who received mini-incision surgery were reviewed retrospectively.The subjects included 5 adrenal tumor excisions,9 nephrectomy,13 unroofing of solitary renal cyst,4 pyeloplasty.12 pyelolithotomy and ureterolithotomy.Results All surgical procedures were successful in the 43 cases.The length of the incision ranged from 3 to 8 cm.The average operation time was 80 minutes and average blood loss was 100 ml. No patients needed blood transfusion during the operation.No serious complications such as the surrounding organ damage happened.The postoperative hospitalization was 5-7 d. Conclusion Mini-incision approach for urological operation has the advantages of minimal invasion,safety,rapid recovery and no requirement for special equipments.Its easy to be popularized in the primary hospital.

Objective The study aimed to explore the relationship between AIF related pathway and the inju-ring of cultured SGNs (spiral ganglion neurons)by glutamate toxicity,and to find AIF expression and distribution changes in SGNs.Methods SGNs of 40 newborn rats within 3 day were obtained and cultured in vitro.Cultured cells were divided into four groups:the normal control group,10 mM,20 mM and 40 mM glutamate injured group, separately.After 48 h hours culturing,optical microscopy,immune fluorescence staining and real-time fluores-cence quantitative PCR were used to observe the morphology,AIF distribution,and AIF,calpain,Caspase3 expres-sion changes in SGNs in vitro.TUNEL was used to verify the cell apoptosis.ResuIts Noticeable morphological chan-ges and cell apoptosis were occurred in 20 mM glutamate group,with AIF nuclear translocation.AIF gene expression was significantly higher than normal after glutamate administration (P<0.05);moreover,calpain gene expression increased(P<0.05);but caspase3 expression was not statistically significantly increased in all glutamate treated groups (P>0.05). ConcIusion In the process of cultured SGNs injured by glutamate,AIF participated in the cell apoptosis.Noticeable cell apoptosis were occurred in 20 mM glutamate group with AIF nuclear translocation.Calpain up-expression also contributed to excitatory neurotransmitter injury on SGNs,but Caspase 3 had no obvious effects.

Objective To study the immune responses of Th1 and Th2 subsets of T cells in 26 children with mycoplasma pneumo-niae pneumonia(MPP).Methods ELISA was used to detect the levels of interferon- ?( IFN- ?) and interleukin-4(IL-4) in the serum in 12 healthy children as normal controls and 26 patients of acute stage as acute stage MPP group, 9 of whom in recovery stage were as recovery stage MPP group. Results IFN - ? level in acute stage MPP group was significant higher than that in normal controls (P 0.05); and IFN - 7/IL - 4 ratio was significant higher than that in normal controls( P0.05).Conclusion Thl responses increase and Th2 responses decrease in children with MPP,and this kind of response persists during the recovery stage.

Objective To improve the recognition of intramedullary spinal abscess by a case of congenital dermal sinus with intramedullary spinal abscess and reduco the incidence of congenital dermal sinus with intramedullary spinal abscess.Methods Clinical,laboratory data and image of a confirmed case about one infant of congenital dermal sinus with multiple intramedullary spinal abscess were investigated,the related literature was reviewed.Results In this case,when the infant with congential dermal sinus had infection,he failed to gain antibiotic therapy, timely surgical treatment,his infection had diffused, and multiple intramedullary spinal abscess flared up.Conclusions Intramedullary spinal abscess is a rare disease.If treatment is delayed, the prognosis is poor and the mortality rate is high.MRI is the ideal investigation for diagnosis.Intramedullary spinal abscess can happen subsequent to congenital dermal sinus with infection, and cause neurological sequela. So an infant with congenital dermal sinus should be offered to avoid complication caused by infection.

A rapid and convenient ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS) method for analysis of ribavirin and its two main kinds of metabolites, 1,2,4-triazole-3-carboxamide (TCONH2 ) and 1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxylic acid (RTCOOH), in fresh eggs has been developed and validated. The sample was extracted with acetonitrile water (9 ∶ 1, V/ V), added N-hexane to move liposoluble substances, after centrifugation, the upper solvent was cleaned-up by C18 and GCB, then determined by UPLC-MS / MS coupled with Agilent ZORBAX SB-Aq column (100 mm×3. 0 mm, 1. 8 μm). Over the concentration range of 2. 0-200. 0 μg / L (0. 5-200. 0 μg / L and 5. 0-200. 0 μg / L) for ribavirin (TCONH2 and RTCOOH) in fresh egg matrix, the correlation coefficients (R2 ) of the calibration curves were above 0. 99, and the limits of detection (LODs) were 0. 54 μg / L (0. 09 and 1. 54 μg / L). The limits of quantitation (LOQs) were 1. 79, 0. 31 and 5. 13 μg / L, respectively. At three spiked concentration levels of 5. 0, 10. 0 and 50. 0 μg / L, the recoveries of ribavirin and RTCOOH ranged from 96. 1% to 99. 6%and 42. 9% to 58. 3% . And at three spiked concentration levels of 0. 5, 2. 0 and 5. 0 μg / L, the recoveries of TCONH2 ranged from 75. 9% to 106. 7% . The results of the determination in various real samples showed that the method is simple, accurate, rapid and suitable determination of ribavirin and its two main kinds of metabolites in fresh eggs.

Objective To investigate the clinical efficacy of a new minimal incision suture technique in repair of acute close rupture of Achilles tendon.Methods From August 2008 through October 2014,134 patients with acute close rupture of Achilles tendon were treated with a self-designed minimally invasive suture technique at our institution.They were 130 men and 4 women,with an average age of 37.4 years (range,from 18 to 52 years).The left side was injured in 93 patients and the right side in 41.The interval from injury to surgery was 1 to 7 days (average,2.5 days).The tendon rupture was located at 4.5 to 6.3 cm proximal to its insertion in 133 patients,and at 1.2 cm in one.With the ankle held in full plantar flexion,a 1.0 to 1.5 cm transverse incision was made over the palpable defect.Forceps were then used to mobilize the tendon from beneath the paratenon.The safe puncture site was determined by course and distribution of the sural nerve indirectly located by magnetic resonance imaging (MRI).Ethicon MB66 nonabsorbable suture was passed diagonally through the tendon bulk under epidural puncture needle guidance.The proximal and distal ends of the suture were tied together and buried.The tendon ends were apposed through the transverse incision.Postoperative individual rehabilitation was established based on pathological differentiation for tendon tissue.Results All the patients were followed up for 8 to 48 months (average,19.6 months).All the wounds healed at the first stage.No complications happened related to incision.The nervus suralis was injured in 3 patients;tendon re-rupture occurred in 4 patients,3 of whom received re-operation and one of whom conservative treatment.Their American Orthopaedic Foot and Ankle Society (AOFAS) scores averaged 99.6,giving a good to excellent rate of 100％ (133excellentcasesandonegoodcase).According to the Arner-Lindholm evaluation criteria,ankle function was excellent in 99 cases,good in 34 ones and poor in one,giving a good to excellent rate of 99.3％.Conclusion The new minimal incision suture technique is worthy of promotion in repair of ruptured Achilles tendon,because it is simple and reliable,does not interfere with the blood circulation to the Achilles tendon,prevents such complications as sural nerve injury and recurrent rupture,and leads to little adhesion and quick functional recovery.