BACKGROUND:Currently,there is not a standard method for in vitro culture and large scale amplification of umbilical cord blood mesenchymal stem cells(UCB-MSCs).OBJECTIVE:To investigate the isolation,purification and culture of UCB-MSCs in vitro,and to detect its surface marker variation.METHODS:The monocytes were harvested from UCB using 1.077 g/cm3 lymphocytes separating solution and density gradient centrifugation,followed by incubation in an incubator containing 5%CO2 at 37℃.The cell morphological changes were observed at different time points and the expression of surface marker was detected using flow cytometry.RESULTS AND CONCLUSION:The monocytes isolated from the UCB grew initially into numerous hematopoietic cell clones,most of which were granulocyte/macrophage colony-forming units and burst forming unit-erithroid,increasing by(37.1±2.3)and (10.4±1.7),respectively.Switzerland staining showed most of them were granulocyte clones(80,1±85.2)%,next was erythroid clones(14.2±1.8)%.At 7 days after culture,some shuttle fibroblast-like cells and fiat osteogenic-like cell spread the whole plastic well.At 14 days after culture,flow cytometry showed CD38+ cells accounted for 1.64%,and CD34+/CD38+ cells accounted for 1,71%,and CD34+/CD38- were 0.55%.PI+ and Annexin-V+ cells accounted for 0.05% and 0.18% respectively.At 21 days after culture,CD38+,CD34+/CD38+ and CD34+/CD38- cells were 74.32%,1.61%,and 0.24%.The results reveled that UCB-MSCs can be isolated and cultured in vitro.

Aim To observe the changes of endothelin receptors and their subtypes of left ventricules in normal SD rats and dilated cardiomyopathy rats. Methods To establish the best conditions of the binding experiment, different protein concentrations, incubation temperature ?and?incubating?time?were?tested? with 125 I-ET-1 ligand respectively. With the selected conditions, saturation binding experiments were performed to determine the amount of endothelin receptor and its subtypes in normal SD rats and in dilated cardiomyopathy ones. Results (1) The optimal incubating temperature was 37 ℃. Under this condition, the binding amount of 125 I-ET-1 increased rapidly in 0～30 minutes, and reached to saturation point at 60 minutes, and there was a linear correlation between 125 I-ET-1 binding amount and cell membrane protein concentration. (2) Endothelin-1, bosentan,BQ123,BQ788 etc. could competitively suppress the bound of 125 I-ET-1 to endothelin receptors. (3) The amount of endothelin receptor in left ventricle of dilated cardiomyopathy rats was ( 92.21? 34.34) nmol?kg -1 protein, which was significantly low than that in normal SD ones. There was no change on the ratio of endothelin receptor subtypes A and B. Conclusion 125I-ET-1 can be used to determine the amount of endothelin receptor and its subtypes in varied tissues specifically. The amount of endothelin receptor in left ventricle of dilated cardiomyopathy rats is down regulated, but the ratio of endothelin receptor subtype A vs B remains to be 21.

Objective To investigate the imaging characters of abdominal cocoon.Methods Six cases of abdominal cocoon proved by surgery and pathologic findings were retrospectively analyzed. Abdominal plain X-ray and CT were performed in 6 cases.The gastrointestinal barium meal series were undergone in 4 cases.The imaging findings were analyzed.Results Abdominal plain X-ray suggested intestinal obstruction in 3 of 6 cases.The gastrointestinal barium meal showed"cauliflower sign"or "concertina pattern"in all of the 4 cases;CT images revealed a conglomeration of multiple small bowel loops in all 6 cases and the intestinal loops seemed to be encapsulated in a membranelike sac.Conclusion The imaging features of gastrointestinal barium meal and CT scan could suggest the diagnose of abdominal cocoon.

Recent years, China has released a series of policies to encourage drug research and development in pediatric populations, aiming to meet pediatric populations' medical needs. Because of the physical and psychological developmental characteristics, tolerance of aversive feelings when taking medications are different between pediatric population and adults. So pediatric populations are at a relatively higher risk of not taking medications as prescribed when the medication tasted unpleasant. Therefore, sound design and evaluation of oral sensory features have important clinical significance and value in developing pediatric medications. "Technical guidance for the design and evaluation of the oral sensory features of pediatric drugs (trial version) " was released in November 2022, by Centre for Drug Evaluation, National Medical Products Administration of China. Based on the guidance, this article will introduce the drafting background and review considerations, hoping to provide reference for the design and evaluation of oral sensory features, and promote drug developing in pediatric population.

Objective To explore the predictive value of brain lobe location of stroke lesion to development of long-term post-stroke depression(PSD)in the first-episode ischemic stroke patients for providing evidence for early intervention.Methods In the prospective study,158 patients aged 60 and over with first-episode ischemic stroke were continuously admitted into Department of Neurology of Shangqiu First People's Hospital from January 2013 to July 2013.The 2 to 3 years follow-up after stroke episode was performed in 126 cases for inquiring into correlation between brain lobe location of stroke lesion and development of PSD.The diagnosis of depression was in accordance with Mental Disorders Diagnostic and Statistical Manual 4(DSM-Ⅳ)standard,and divided into groups of stroke with depression(n=52)and stroke without depression(n=74).The degree of depression was evaluated by 17-item Hamilton Rating Scale for Depression(HAMD17).The location,number and volume of stroke lesions were determined by head MRI.The relationship between PSD and pathogenetic loci was analyzed by unconditional Logistic regression analysis.A difference in hemisphere commensalism.Results The morbidity of PSD was 41.3%(52/126),with 21.4%(27/126),12.7%(16/126),7.1%(9/126)in mild,moderate and severe PSD respectively.The frontal lobe(OR=2.824,95%CI=1.189-6.706)and the temporal lobe(OR=3.579,95%CI=1.233-10.393)cerebral infarction were correlated with the occurrence of PSD.The long-term PSD severity was more in frontal lobe than in temporal lobe(χ2=6.399,P<0.05).The average volume of cerebral infarction was larger in PSD group than in non-PSD group(t=3.271,P<0.05),and the average number of cerebral infarction loci was more in PSD group than in non-PSD group(t=3.176,P<0.05).The more severe the degree of depression according to HAMD17,the larger the average volume of cerebral infarction(F=6.280,P<0.05)and the more the average number of lesions(F=6.132,P<0.05).Conclusions The development of long-term PSD in the first-episode elderly patients is affected by the invasion site.The frontal lobe and temporal lobe infarction are independent risk factors for long-term PSD in patients with ischemic stroke,and the PSD was more severe in frontal lobe infarction than in temporal lobe infarction.

Objective To study the differences of esophageal manometry (M),pH set-up method (P) and X-ray (X) on positioning of the 24-hour esophageal pH catheter and relative factors. Methods Fifty subjects underwent M, then pH catheter was located by P and X. The difference between methods and the relative factors such as body height,lower esophageal sphincter (LES) length etc were analyzed. The difference of less than 2 cm between two methods was accepted. Results The length between the location determined by M,Pand X and nose were (37.9 ±2.9),(40.8 ±4.4) and (40.7 ±5.3) cm, respectively.There was significant difference between P and M as well as between X and M (P＜ 0.01 ). The coincidence rate was 62.0%(31/50) between P and M;84.0%(42/50) between P and X;and 58.0%(29/50) between X and M. Compared with P,M was (2.9 ±3.9) cm nearer to the stomach. Age, body height and LES length were main factors which affect the difference between P and M (P＜ 0.01 or ＜ 0.05),body height and LES length were main factors which affect the difference between X and M (P＜0.05 or ＜0.01). Conclusions Compared with M, the location determined by P is nearer to stomach. The location of X is varied. Body height and LES length are main relative factors.

Objective To explore the clinical characteristics of and to provide help to the prevention and treatment of malignant chest tumor with nosocomial mixed fungal-bacterial infection.Methods From July 2007 to June 2015,pathogenic bacteria in sputum,blood,urine,chest incision,thoracic and abdominal fluid,and implantable medical biological material were cultivated in 5067 patients with malignant chest tumor suspected with infection.The clinical characteristics,source of specimen and pathogenic bacteria,the types of diseases,medical intervention activities of 142 cases detected with mixed fungal-bacterial infection were retrospectively analyzed.Fesult In 142 patients,104 patients at clinical stage Ⅲ-Ⅳ accounted for 73.2％,and 94 patients used antibiotics more than 14 days (66.2％);104 cases had implanted biological materials (74.7％);96 cases died (67.6％).A total of 167 strains bacteria were isolated.Sixty-one strains of G+ bacteria accounting for 36.5％ were mainly Epidermis staphylococcus and Staphylococcus aureus;106 strains of G-bacteria accounting for 63.5％ were mainly klebsiella pneumonia,Escherichia coli and baumanii;172 strains fungus mainly of Candida albicans were isolated (77.3％).Pathogenic bacteria sources were mainly sputum specimens + pharynx strip,blood culture and medical implant materials.In 72 lung cancer patients,squamous carcinoma and small cell carcinoma were 52.8％ and 33.3％ respectively,higher than adenocarcinoma (12.5％);In 42 esophageal cancer patients,postoperative patients were 42.9％.Parenteral nutrition patients with more than 10 days were 80.9％ higher than that of parenteral nutrition in patients with less than 10 days (19.1％).Conclusion Among malignant chest tumor patients with nosocomial mixed fungal-bacterial infection,the bacteria were found in staphylococcus aureus,klebsiella pneumoniae and E.coli and the fungus was Candida albicans.For clinical stage Ⅲ-Ⅳ,patients with parenteral nutrition for more than 10 days,having history of chemo or radiotherapy,with antimicrobial use for more than 14 days,and with implanted biological materials,should be warned about nosocomial mixed fungal-bacterial infection.

Objective To detect and analyze the mutation of fibroblast growth factor receptor 3(FGFR3) gene among a family with congenital achondroplasia(ACH).Methods Six blood samples of family member in this pedigree were cellected according to the informed consent process for genetic research,and 2 unralted healthy human blood sample were taken as controls.The mutation at nucleotide position 1 138 on FGFR3 gene was detected by using Polymerase chain reaction and single-strand conformation polymorphism(PCR-SSCP)and polyme-rase chain reaction and restriction endonuclease technology(PCR-RFLP) methods.Results Using PCR-SSCP method firstly,only the proband with ACH and his father in this family had the same abnormal band.The amplified products including 1 138 loci on FGFR3 gene further was analyzed by Sfe Ⅰ digestion,3 fragments including 164 bp,109 bp and 55 bp were detected in the proband and his father again,and the other members in the family and 2 controls just showed 164 bp band.It indicated that just 2 patients (proband and his father) showed heterozygous G→A transition mutation at nucleotide position 1 138 on the FGFR3 gene.The amplified products at 1 138 loci was also detected by MspⅠ digestion,just 1 band was observed in all members in this family and 2 controls.It showed that there was no G→C substitution at nucleotide position 1 138.Conclusions The G→A transition mutation at nucleotide position 1 138 in transmembrane domain of FGFR3 gene may be the main cause of achondroplasia in this family.In this pedigree,the proband showed's father a de novo mutation which was transferred to his child again.

In industrial glutamate fermentation, intermitted feeding glucose with the help of off-line glucose measurement is generally necessary. This kind of feeding strategy could cause large variations in glucose concentration so that it is not favorable for the achievement of efficient and stable glutamate fermentation. Glutamate fermentation is characterized with typical non-growth association behavior, and during glutamate production phase glucose consumption is closely correlated with ammonia consumption. In this study, glucose concentration was controlled at various pre-determined levels by predicting glucose consumption amount and thus its concentration with the aid of on-line monitoring ammonia consumption. When glucose concentration was controlled around a lower level of 5 g/L~10 g/L, the final glutamate concentration could reach a relatively higher level of 80 g/L. In this way, the huge osmotic stress change due to the large glucose concentration variation with the intermitted feeding method could be avoided and the glutamate fermentation performance enhancement be expected.

ObjectiveTo observe the effect of ultrashort waves combined with medication on children with cerebral dysfunction syndrome (CDS) accompanied with pneumonia.Methods76 children with CDS accompanied with pneumonia were randomly divided into treatment group (n=38, treated with anti-inflammatory and ultrashort waves one time per day for ten days) and control group (n=38, treated only with anti-inflammatory). The effective rate and the days of rales vanished of children in two groups after treatment were compared.ResultsIn the treatment group, 14 cases cured, 20 cases got significant result, 4 cases were effective, total effective rate was 100%; in the control group, 10 cases cured, 14 cases got significant result, 9 cases were effective, 5 cases were invalid, total effective rate was 86.8%. There was a significant difference between therapeutic effects of two groups ( P<0.05).ConclusionThe ultrashort waves therapy is an effective method for the children with CDS accompanied with pneumonia.