BACKGROUND: Researches show that erythropoietin (EPO) can stimulate the proliferation and protraction of endothelial progenitor cells to form new vessels, so EPO may play an important role in proliferation and differentiation of bone marrow-derived mesenchymal stem cells (MSCs). OBJECTIVE: To explore the effects of EPO on the proliferation and cell cycle of MSCs. DESIGN, TIME AND SETTING: From July to November 2007, the observation of comparative cell trial was performed at the Hematologic Diseases Institute of General Hospital Lanzhou Military Area Command of Chinese PLA. MATERIALS: Bone marrow liquid specimens were provided by voluntary donors. The informed consents were obtained from all patients, and the experiment was approved by Hospital Ethics Committee. METHODS: Using Percoll solution, MSCs were isolated from bone marrow by the method of density centrifugation. Flow cytometry was applied to detect the cell cycles of the second and third passages. MSCs were incubated with different doses of EPO (0.25, 0.50, 1.00, 2.00, 5.00 U/mL) in serum free culture media, and cells cultured with no EPO were regarded as control. All cells were cultured for 24, 48, and 72 hours. MAIN OUTCOME MEASURES: ①Proliferation of MSCs was measured by MTT assays after 24, 48, and 72 hours; ②Cell cycle was measured by flow cytometry assays after incubated with 10 U/mL EPO for 72 hours. RESULTS: After MSCs was incubated with EPO, the cell proliferation index was significantly increased in a dose and time dependent manner. The effects on the proliferation of MSCs were highest in 5 U/mL group. Compared with the control group, EPO could significantly decrease G0 /G1 ratio, and increase S and G2/M stage ratio (P

BACKGROUND: No particular marker molecule of bone marrow mesenchymal stem cells (BMSCs) is presently found, so its determination is difficult. BrdU marker has no radioactive pollution. Some studies have confirmed that BrdU marker has no damage to cell function without ultraviolet radiation. OBJECTIVE: To investigate in vitro identification and labeling methods and biological characteristics of adult BMSCs. DESIGN, TIME AND SETTING: The cell experiment was conducted at the General Hospital of Lanzhou Military Area Command of Chinese PLA between June and December 2007. MATERIALS: Bone marrow was collected from 5 healthy adult volunteers. BrdU was purchased from Sigma, USA. METHODS: 10 mL adult bone marrow was harvested to isolate mononuclear cells by density gradient. Cells were cultured in DMEM containing 10% fetal bovine serum and proliferated at a density of 2?108 L-1. At the third passage, BMSCs was inoculated in medium supplemented with osteoblast inductor and stained with alkaline phosphatase. Subsequently, BMSCs were inoculated in medium supplemented with lipoblast inductor and stained with oil red O. Cells were incubated with BrdU at different concentrations of 5, 10 and 15 ?mol/L for 12, 24, 48, 72 and 96 hours, and then detected by immunocytochemistry. MAIN OUTCOME MEASURES: Cell growth and proliferation were observed under an inverted light microscope. Cell phenotype, osteoblast and lipoblast differentiation were identified by flow cytometry. BrdU-positive labeling rate and cell growth after labeling were investigated. RESULTS: In vitro cultured BMSCs were homogenous population and exhibited a spindle-shaped fibroblastic morphology. BMSCs expressed CD44 and CD71, but did not express CD34 and HLA-DR. BMSCs can differentiate into osteoblasts and lipoblasts. Most BMSCs were labeled by BrdU. With the increase in labeling concentration and over time, BrdU positive rate gradually increased and exceeded 90% after labeling with BrdU (10 ?mol/L) for 72 hours, with the labeling identifiable in nine consecutive passages. At the third passage, 90% BMSCs were in G0/G1 phase, and 88.62% in G0/G1 phase after labeling. BrdU has little effect on morphous and proliferation of labeled cells. CONCLUSION: Adult BMSCs can be identified through morphological character, specific surface antigens and multipotential differentiation in vitro. BrdU labeling provides a feasible means for a dynamic observation of the survival, growth and differentiation of the implanted BMSCs. The optimal dosage and timing of BrdU labeling are respectively 10 ?mol/L and 72 hours.

Objective:To analysis the Mimotopes of the peptide mimics to PGN using online softwares.Methods: Mimotopes of PGN were screened from 12-mers linear phage display peptide library by using anti-PGN McAb and the antigenicity of selected clones was identified by ELISA.The B cell epitopes,T cell epitopes of ′GRWxHxVxWAGL′ were estimated by DNAstar and online softwares.Results: 16 phage clones that bound with anti-PGN McAb were screened from 12-mers linear phage display peptide library.Among these positive clones,phage clones No.39 shared the conserved sequence:WxHx……AGL found in previous clone No.31(ATWxHxLxSAGL),which provoked an effective protective immunity against infection with S.aureus.To enhance the stability of the conformation as well as adding biotin on the N-ter-minal as a tag,the sequences ′39′ were redesigned and synthesized by adding S(serine)A(alanine) and GG(glycine)on the C-terminal of origin sequence(named SP39).Next,we estimated or predicted antigenic epitopes,T cell epitopes and scores binding to MHC of these peptides by using DNASTAR and online softwares(http://bio.dfci.harvard.edu/Tools/antigenic.pl,www.syfpeithi.de,http://www.darrenflower.info/mhcpred),indicating that SP39 contains sites bound both mice and human MHC.The sequences ′WxHxVxW-′ may be antigenic epitope as SP39,which contains a T cell epitope.Our results showed that both SP39 could bind to both anti-PGN McAb and a polyclonal antibody against S.aureus.Moreover PGN could inhibit the binding of SP39 to the anti-PGN McAb.These data indicated that SP39 mimic to epitopes on PGN.Conclusion: SP39(GRWxHxVxWAGLAGGS) probably display the mimotopes of PGN.

Objective:To prepare the polyclonal antibodies against advanced oxidation protein products (AOPP),and to provide an effective agent for research on the pathogenesis of AOPP and assess exactly the relationship between AOPP and relative diseases.Methods:AOPP-rabbit serum albumin (AOPP-RSA) was prepared by treating RSA with hypochloric acid.The rabbit anti-AOPP-RSA polyclonal antibodies were generated and purified by affinity chromatography. The titers and the specificity of the antibodies were measured by ELISA.The plasma AOPP and the localization of AOPP in nephridial tissues of some patients with chronic kidney disease (CKD) were determined using rabbit anti-AOPP-RSA.Results:Titers of the antibodies were 10-6.Purified antibodies reacted specifically with oxidized albumin from different genus,but could not react with normal albumin and glycosylated albumin.The high level of AOPP in plasma from CKD patients was confirmed by Western blot.The antibodies could be used to immunostain AOPP deposition in different regions of kidney tissues from both CKD patients and CKD rat models.Conclusion:We successfully generate rabbit anti-AOPP polyclonal antibodies with high titers and striking specificity.The presence of plasma AOPP and localizations of AOPP in kidney tissues of CKD patients can be demonstrated using the antibodies.The development of anti-AOPP polyclonal antibodies may provide a new tool to explore the pathogensis of AOPP and assess exactly the relationship between AOPP and relative diseases.

Objective:To screen epitope mimics to lipoteichoic acid from a random 12-mer phage display peptide library and i-dentify the specificity of the mimotopes of LTA.Methods:The monoclonal antibody against LTA was used as a target to screen the 12-mer phage display peptide library and the specificity of phage clones were identified by sandwich ELISA.The amino acid sequences of positive phage clones were deduced from DNA sequencing.The specificity of synthetic peptide were identified by sandwich ELISA.Results:4 clones were obtained after 3 rounds of screening.Amino acid sequence analysis revealed four different types of mimotope sequence.A linear peptide (GHxDFRQxxQPS),named L2,which derived from positive sequence was synthesized.ELISA result indicates that L2 can bind to anti-LTA mAb specifically in a dose-dependent manner.Conclusion:The mimotopes of LTA were obtained by using the phage display technology.

Objective:To evaluate the anterior expansion of sacral foramen and decompression of sacral plexus via the lateral-rectus approach (LRA) in the surgical treatment of sacral fractures complicated with sacral plexus injury.Methods:From January 2013 to June 2018, 11 patients were treated at Department of Orthopaedics, The Third Hospital Affiliated to Southern Medical University for obsolete sacral fractures complicated with sacral plexus injury. They were 8 males and 3 females, aged from 17 to 54 years (average, 38 years). According to the Denis classification, all the sacral fractures belonged to Denis Zone Ⅱ. According to British Medical Research Council (BMRC) grading system, the nerve injury was complete damage in 2 cases and partial damage in 9. The mean time from injury to surgery was 6 months (range, from 0.7 to 12.0 months). After the sacroiliac joint was exposed via the LRA, the lumbosacral trunk was exposed and released between iliac vessels and the iliopsoas. Next, the S1 foramen was expanded and the S1 nerve root was released after separation of the median sacral artery and the internal iliac artery. Reduction and fixation of the sacroiliac joint was carried out for patients with unstable sacral fracture. X-ray and CT examinations of the pelvis were performed to evaluate fracture healing and neurological function recovery postoperatively.Results:Of this cohort of 11 cases, operation succeeded in 10 but failed in one whose sacral fracture was found to have completely healed with the S1 foramina totally occluded. The surgical time averaged 110 min (range, from 70 to 220 min) and the blood loss 1, 100 mL (range, from 450 to 2, 800 mL). Postoperative X-ray and CT examinations showed that the sacral foramens were expanded significantly without any complications. The follow-up time averaged 18 months (range, from 12 months to 4 years). By the BMRC grading system at the last follow-up, the neural function was completely recovered in 5 cases, partially recovered in 4 cases and not recovered in one.Conclusion:Significant anterior expansion of sacral foramen and decompression of sacral plexus via the LRA is a viable and effective alternative for treatment of sacral fractures complicated with sacral plexus injury.

Objective To investigate the prevalence of infantile eczema in Uygur and Han nationalities and the relationship between pregnancy factors and 0-6 months infant eczema, so as to provide a scientific basis for formulating preventive strategies. Methods From January-August 2016 in the Xinjiang Autonomous Region People's Hospital of obstetric and follow-up to fill out the questionnaire in 570 pregnant women and their babies as the research object to establish the baseline survey cohort, pregnancy related information to pregnant women, and 6 months (January, March, on maternal June) follow-up, observe the occurrence of infantile eczema a total of 502, successful follow-up to maternal and child. Using nested case-control study, the cohort occurred in 182 cases of infant eczema as case group, will be in the queue without eczema in 320 infants as control group, using multivariate logistic regression analysis of risk factors during pregnancy baby eczema. Results 0-6 months infant eczema incidence rate was 36.25％, the incidence of Uygur infant eczema (20.43％) was significantly lower than that of Han nationality infant eczema incidence (50.19％) (OR=0.45,95％CI:0.35-0.69) P<0.01). Social and economic factors affecting the occurrence of infantile eczema comparison results showed that the ethnic factors and infant eczema, and more prone to eczema than that of Uygur nationality; and Uygur compared mothers influence allergic history of Han infantile eczema more (χ2=7.09, P=0.008). Multivariate logistic regression analysis showed that the mother has a history of atopy (OR=7.17,95％CI:4.15-12.37), mother′s education level (OR=1.91,95％CI:1.51-2.74), the higher the monthly income (OR=2.68,95％CI:2.17-3.33), maternal smoking during pregnancy (secondhand smoke) (OR=2.42,95％CI:1.96-2.97), frequently eating spicy spicy foods during pregnancy (OR=1.94,95％CI: 1.41-2.68) will increase the risk of infant eczema Supplementation of probiotics (OR=0.13,95％CI:0.07-0.25) and vitamin D (OR=0.13,95％CI:0.06-0.26) during pregnancy is a protective factor in the development of infantile eczema. Conclusions Compared to Uygur, Han Chinese infants are more likely occur eczema, and family history of allergies, education, economic income, pregnancy dietand smoking are risk factors for infant eczema. The same risk factors among Uygur and Han ethnic ,but it had a different results. Compared to Han, Uygur pregnant women eating spicy food caused the risk of infant eczema occurrence is higher, but the short period of housing renovation caused the risk of infant eczema occurrence is lower. Therefore, it is necessary to strengthen nutrition andmonitor living environment during pregnancy. Avoiding to eat spicy food and supplying probiotics and vitamin D, it can prevent or reduce the occurrence of infantile eczema.

Objective To analyze the pregnancy outcomes of fetal tetralogy of Fallot and to explore its prenatal diagnosis and treatment procedures. Methods The clinical data of 63 cases of fetal tetralogy of Fallot (62 cases were singleton and 1 case was one of twin) were collected retrospectively from November, 2013 to November, 2017 in Beijing Obstetrics and Gynecology Hospital. Results (1) Totally, 63 cases out of 46 352 pregnancies were diagnosed fetal tetralogy of Fallot by fetal ultrasonic cardiogram with about 0.136%(63/46 352) occurrence rate, and the mean gestational age was (23±3) weeks. And 50 cases (79%, 50/63) terminated pregnancy by induced labour. (2) Totally, 57 cases (90%,57/63) accepted genetic diagnosis.Eight cases (13%, 8/63) existed chromosome abnormality including 21-trimosy in 6 cases, 18-trisomy in 1 case and 22q11.2 microdeletion syndrome in 1 case; and these 8 cases were determined before 28 gestational weeks. (3) And 13 cases (21%, 13/63) of no fetal genetic abnormality selected to continue pregnancy. Twelve cases underwent full term delivery (5 cases were cesarean section delivery and 7 cases were vaginal delivery). Twelve newborns underwent surgical radical operation on heart malformation and got recovery. One case underwent preterm cesarean section at 35 gestational weeks for one of twin, and the newborn with tetralogy of Fallot was dead. The other the newborns survived and were followed up for tetralogy of Fallot surgery from 1 month to 3 years old after birth and recovered.Conclusions Fetal tetralogy of Fallot mainly is diagnosed by ultrasonic cardiogram in the second trimester. The gestational age of diagnosis may be as early as 15 gestational weeks. Fetal tetralogy of Fallot with no genetic abnormality could underwent radical heart malformation operation after birth. It is necessary to undergo genetic testing on fetal tetralogy of Fallot and prenatal multidisciplinary counseling as well.

Objective: To analyze the pregnancy outcomes of fetal tetralogy of Fallot and to explore its prenatal diagnosis and treatment procedures. Methods: The clinical data of 63 cases of fetal tetralogy of Fallot (62 cases were singleton and 1 case was one of twin) were collected retrospectively from November, 2013 to November, 2017 in Beijing Obstetrics and Gynecology Hospital. Results: (1) Totally, 63 cases out of 46 352 pregnancies were diagnosed fetal tetralogy of Fallot by fetal ultrasonic cardiogram with about 0.136%(63/46 352) occurrence rate, and the mean gestational age was (23±3) weeks. And 50 cases (79%, 50/63) terminated pregnancy by induced labour. (2) Totally, 57 cases (90%,57/63) accepted genetic diagnosis.Eight cases (13%, 8/63) existed chromosome abnormality including 21-trimosy in 6 cases, 18-trisomy in 1 case and 22q11.2 microdeletion syndrome in 1 case; and these 8 cases were determined before 28 gestational weeks. (3) And 13 cases (21%, 13/63) of no fetal genetic abnormality selected to continue pregnancy. Twelve cases underwent full term delivery (5 cases were cesarean section delivery and 7 cases were vaginal delivery). Twelve newborns underwent surgical radical operation on heart malformation and got recovery. One case underwent preterm cesarean section at 35 gestational weeks for one of twin, and the newborn with tetralogy of Fallot was dead. The other the newborns survived and were followed up for tetralogy of Fallot surgery from 1 month to 3 years old after birth and recovered. Conclusions Fetal tetralogy of Fallot mainly is diagnosed by ultrasonic cardiogram in the second trimester. The gestational age of diagnosis may be as early as 15 gestational weeks. Fetal tetralogy of Fallot with no genetic abnormality could underwent radical heart malformation operation after birth. It is necessary to undergo genetic testing on fetal tetralogy of Fallot and prenatal multidisciplinary counseling as well.