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.

OBJECTIVETo investigate clinical characteristics of posterior cruciate ligament (PCL) injuries.

METHODSTo review all 326 patients with PCL injuries from January 2006 to June 2013 retrospectively, which were grouped according to injury patterns. The survey included the gender, age, sides, duration from injury to surgery, cause of injury and combined injuries. Compare the patient-specific factors between two groups.

RESULTSMale patients were the majority of PCL injuries (73%), and 30 to 50 years old patients accounted for 59%. 62.6% of all patients went for a doctor within one month after trauma. Traffic accidents were the main cause of PCL injuries (59%), especially motorcycle accidents (74.3%). Anterior cruciate ligament (ACL) injuries were the most common combined injuries (46.9%),followed by medial collateral ligament (MCL) (29.8%) and posterolateral corner (PLC) (26.1%). The meniscal tears accounted for 30.4% of total cases. The in-substance PCL injuries (71.5%) were far more than avulsion fractures (28.5%), and the former had more combined injuries than the later (P < 0.05). The causes of injuries were statistically different between two groups (P < 0.05). The prevalence of isolated avulsions was higher than isolated in-substance injuries (P < 0.05).

CONCLUSIONIn the general population, in-substance PCL tears were dominant on PCL injuries. The prevalence of combined injuries was quite high, among which ACL injuries were the most common. Insertion avulsion fractures emerged mainly at the tibial side, which had less combined injuries than those suffered in-substance PCL injuries. The incidence of isolated insertion avulsion fractures was higher than isolated in-substance injuries.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Female ; Humans ; Incidence ; Knee Injuries ; epidemiology ; Male ; Middle Aged ; Posterior Cruciate Ligament ; injuries ; Retrospective Studies ; Young Adult

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.

Objective To investigate the relationship between abnormal methylation in promoter regions for OPN and VSMC phenotype switching in varicosity.Methods Immunohistochemistry and Western-blot were used to evaluate the expression of SMA and OPN in VSMC.Methylation-specific PCR was used to evaluate the methylation level of OPN in VSMC of vein samples.Ultrastructure change of VSMC was observed by transmission electron microscope (TEM).Results Compared to normal vein,OPN in VSMCs were significantly highly expressed,mainly in the neointimal region (P ＜ 0.01).SMA in neointima region was in low expression (P ＜ 0.01).The density of OPN in varicose group was significantly higher (P ＜0.01).DNA methylation level of OPN was lower in varicose veins.Conclusions Hypomethylation of the promoter regions for OPN may cause high expression of OPN leading to VSMC phenotype switching and development of varicosity.

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.