Objective To study the characteristics of mesenchymal stem cells(MSCs)derived from chronic myelogenous leukemia(CML)patients' bone marrow and examine their hematopoiesis support in vitro.Methods MSCs from CML were obtained and cultured.Immunophenotype and in vitro differentiation capacities were investigated.Moreover,the Ph chromosome and bcr/abl gene of CML derived MSCs were detected.The expression of cytokines was detected by RT-PCR,and hema- topoiesis support of MSCs in vitro was detected by long-term bone marrow culture and the methylcel- lulose progenitor assay.Results CML derived MSCs showed a typical fibroblast like morphology. They were positive for CD29,CD44 and CD105,while negative for CD14,CD31,CD45,CD34 and HLA-DR.Under suitable conditions,CML derived MSCs could differentiate into osteoblasts and adi- pocytes.Further,CML derived MSCs showed a normal choromosome,and did not express bcr/abl gene.At last,they expressed hematopoiesis cytokines and possessed ability of hematopoiesis support in vitro.Conclusion There are MSCs with multidirectional differentiation potential in CML bone mar- row and they possess the ability of hematopoiesis support.

Mesenchymal stem cells (MSCs) are promising candidates to develop new cell-based therapeutic strategies and bone marrow represents the main source of MSCs for experimental and clinical application currently. How to obtain enough MSCs is the biggest challenge faced by the experimental and clinical application. The difference of morphology, growth pattern, immunophenotype and multilineage differentiation of MSCs derived from human umbilical cord vein and their multiple differentiation capacity were investigated. The results showed that MSCs derived from human umbilical cord vein and normal adult bone marrow were similar in biological characteristics. They not only grew in fibroblast-like cells mode but had great expansion potency and multilineage differentiation ability in vitro. The MSCs derived from human umbilical cord vein may be an excellent alternative source for experimental and clinical application.

Objective: To explore the application value of optical coherence tomography angiography (OCTA) in retinal microvascular blood flow in children with anisometropic amblyopia. Methods: 31 children with monocular anisometropic amblyopia in the Department of Ophthalmology in Shanghai General Hospital, Shanghai Jiao Tong University from June to August in 2018 were included. The 3 mm × 3 mm and 6 mm × 6 mm area centered on the macular area and 4.5 mm × 4.5 mm area centered on the optic disc area were detected by OCTA. The software automatically calculated the average blood flow densities of superficial capillary plexus (SCP) and deep capillary plexus (DCP), and the average blood flow densities of optic disc and the peripapillary area, the average area of foveal avascular zone (FAZ), as well as the average foveal thickness in the anisometropic amblyopia and the contralateral eyes. The anisometropic amblyopia eyes of the children were classified as the amblyopia group, and the contralateral non-ambly eyes were classified as the contralateral eye group. The differences in blood flow parameters between two groups were compared. Results: In the 3 mm × 3 mm macular scan, the average blood flow densities of SCP and DCP in the amblyopia group were (46.40±4.72)% and (52.17±2.82)%, respectively, and the average blood flow densities in the contralateral eye group were (48.48±3.46)% and (54.31±2.18)%, respectively. The differences in the average blood flow densities of SCP and DCP between two groups were statistically significant (P=0.012, P=0.012). In the 6 mm × 6 mm macular scan, the average blood flow densities of SCP and DCP in the amblyopia group were (47.41±3.04)% and (48.92±5.34)%, respectively, and the average blood flow densities in the contralateral eye group were (50.36±2.70)% and (51.54±4.69)%, respectively. The differences between two groups were statistically significant (P=0.016, P=0.046). In the 4.5 mm × 4.5 mm optic disc scan, the average blood flow densities of the optic disc in the amblyopia group and the contralateral eye group were (48.98±4.03)% and (52.06±3.90)%, respectively. The difference was also statistically significant (P= 0.040). The average blood flow densities of the peripapillary area in the amblyopia group and the contralateral eye group were (52.16±2.22)% and (52.44±1.50)%, respectively. The difference was not statistically significant. Conclusion: The average blood flow densities of SCP, DCP and optic disc in the anisometropic amblyopia eyes are significantly lower than that in the contralateral eyes. OCTA has certain application value in evaluating retinal blood flow in children with anisometropic amblyopia.

Objective To explore the impact factors for post-thaw embryo survival rate and clinical pregnancy rate in frozen-thawed embryo transfer program. Methods The clinical data of 573 cycles of frozen-thawed embryo transfers were retrospectively analysed. Groups were divided according to the pre-freeze embryo quality, pre-freeze embryonic developmental stage, frozen-thawed embryo quality and cryopreservation technique, respectively, and post-thaw embryo survival rates and/or clinical pregnancy rates were compared among groups. Results The clinical pregnancy rate of high quality pre-freeze embryo was significantly higher than that of low quality pre-freeze embryo (31.8% vs 20.0%) (P< 0.05). There was no significant difference in the post-thaw survival rates and clinical pregnancy rates between embryos frozen at day 2 of ferrtilization and those frozen at day 3 of ferrtilization(79. 1% vs 82.9% and 25.5% vs 31.2%, respectively) (P>0.05). The clinical pregnancy rates of the transfer cycles only with fully intact embryos and with mixed embryos were significantly higher than that only with partially damaged embryos(36.7% vs 24.1% and 29.2% vs 24.1%, respectively)(P<0.05). The post-thaw survival rate and post-thaw high-quality embryo rate were significantly higher in those processed with modified cryopreservation technique than in those processed with original cryopreservation technique (82.0% vs 66.3% and 50.0% vs 27.5%, respectively)(P<0.05). Conclusion Pre-freeze embryo quality, post-thaw embryo survival rate and post-thaw embryo quality have a positive correlation to subsequent clinical pregnancy rate. Favorable cryopreservation technique may ensure the success of post-thaw embryo recovery and transfer.

Objective:To investigate the effects of vascular endothelial growth factor (VEGF) on the expression of secreted protein acidic and rich in cysteine (SPARC) and the fibrosis in cultured human Tenon's fibroblast (HTF) in vitro. Methods:HTF cells were obtained from Tenon's capsule tissues of patients undergoing strabismus surgery. Immunofluorescence was used to identify the HTF cells. HTF cells were cultured with different concentrations of VEGF, and which were divided into four groups, i.e., 0 ng/mL group, 25 ng/mL group, 50 ng/mL group and 100 ng/mL group. The expression of SPARC, collagen- , and matrix metalloprotein 9 (MMP-9) and the activity of extracellular signal-regulated kinase (ERK) pathway were analyzed by Western blotting and real-time quantitative PCR (qPCR). The abilities of proliferation and migration of HTF cells were detected by MTS assay and scratch test, respectively. Results:HTF cells were observed and identified by inverted phase contrast microscope and immunofluorescence. Under the stimulation of VEGF, the expression of protein and mRNA of SPARC, collagen-I and MMP-9 of HTF cells in other three groups were increased compared with 0 ng/mL group; the phosphorylation activities of ERK pathway were up-regulated, and the proliferation and migration abilities of HTF cells were up-regulated. And the effect was the most obvious in the 50 ng/mL group. Conclusion:VEGF is involved in promoting the fibrosis of HTF cells accompanied by the up-regulation of the SPARC, which suggests SPARC may become a potential regulatory site.

Objective To explore the clinical effect of Fu's subcutaneous needling combined with western medicine in the treatment of irritable bowel syndrome(IBS).Methods A total of 112 patients with IBS were randomly divided into the observation group and the control group,with 56 patients in each group.The control group was given conventional western medicine treatment,and the observation group was treated with Chinese medicine Fu's subcutaneous needling on the basis of the treatment in the control group.Both groups were treated for 4 consecutive weeks.After 1 month of treatment,the clinical efficacy of the two groups was evaluated,and the changes in the IBS system severity system(IBS-SSS)scores were observed before and after the treatment,as well as the time of the symptoms of abdominal pain,bloating and diarrhoea subsiding in the two groups,and the changes in the defecation thresholds,pain thresholds,sensory thresholds and the changes in the numbers of intestinal flora such as Bifidobacterium,Lactobacillus,Enterobacteriaceae,and Bacteriodendrobacteriaceae in the two groups were compared.The changes in the levels of vasoactive intestinal peptide,gastric motility and substance P were observed in the two groups.The safety and occurrence of adverse reactions were also evaluated in the two groups.Results(1)The total effective rate of the observation group was 96.43%(54/56),and the control group was 83.93%(47/56).The efficacy of the observation group was superior to that of the control group,and the difference was statistically significant(P＜0.05).(2)The time for abdominal pain,abdominal distension and diarrhoea to subside was significantly shorter in the observation group than in the control group,and the difference was statistically significant when compared with the control group(P＜0.05).(3)After treatment,the defecation threshold,pain threshold,and sensation threshold of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in terms of improving defecation threshold,pain threshold,and sensation threshold,and the difference was statistically significant(P＜0.05).(4)After treatment,the numbers of Bifidobacterium,Lactobacillus,Enterobacter,Bacteroides were significantly improved in the two groups(P＜0.05),and the observation group was significantly superior to the control group in terms of improving the numbers of Bifidobacterium,Lactobacillus,Enterobacter and Bacteroides,and the difference was statistically significant(P＜0.05).(5)After treatment,the levels of vasoactive intestinal peptide,gastric actin,and substance P of the patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the levels of vasoactive intestinal peptide,gastric actin,and substance P,and the differences were all statistically significant(P＜0.05).(6)There was no significant difference in the incidence of adverse reactions between the two groups(P＞0.05).Conclusion Fu's subcutaneous needling combined with western medicine in the treatment of IBS can significantly shorten the relief time of patients'clinical symptoms,strengthen patients'gastrointestinal function,regulate the number of intestinal flora and improve the function of gastrointestinal mucous membrane barrier,and the therapeutic efficacy is remarkable.

OBJECTIVETo investigate whether the protective effects of puerarine (Pur) against cerebral ischemia is associated with depressing the extracellular levels of amino acid transmitters in brain of rats.

METHODSMale Sprague-Dawley rats were subjected to transient middle cerebral artery occlusion (MCAO) for 60 min followed by 24 h reperfusion. Pur (50, 100 mg/kg, i.p.) was administered at the onset of MCAO. The infarct rate and edema rate were detected on TTC (2,3,5-triphenyltetrazolium chloride)-stained coronal sections. The extracellular levels of amino acid transmitters were monitored in striatum of rats with ischemic/reperfusion injury using in vivo microdialysis technique. Furthermore, the protective effects of Pur against glutamate-induced neurotoxicity were detected. Glutamate-induced apoptotic and necrotic cells in hippocampus were estimated by flow cytometric analysis of Annexin-V and PI labeling cells.

RESULTSPur (100 mg/kg) significantly decreased infarct size by 31.6% (P<0.05), reduced edema volume (P<0.05), and improved neurological functions (P<0.05) following MCAO. In these rats, the ischemia-induced extracellular levels of aspartate (Asp), glutamate (Glu), y-aminobutyric acid (GABA), and taurine (Tau) were significantly reduced in striatum of vehicle-treated animals by 54.7%, 56.7%, 75.8%, and 68.1% (P<0.01 and P<0.05). Pur reduced the peak values of Glu and Asp more obviously than those of GABA and Tau, and the rate of Glu/GABA during MCAO markedly decreased in Pur-treated MCAO rats, compared with the vehicle-treated MCAO rats. Meanwhile, apoptosis and necrosis induced by Glu in cultured hippocampal neurons were significantly reduced after Pur treatment.

CONCLUSIONAcute treatment with Pur at the onset of occlusion significantly depresses ischemia-induced efflux of amino acids, especially, excitotoxicity in the striatum, a mechanism underlying the neuroprotective effect on cellular survival.

Animals ; Biological Transport ; Brain Ischemia ; pathology ; prevention & control ; Excitatory Amino Acids ; metabolism ; Flow Cytometry ; Hippocampus ; drug effects ; pathology ; Isoflavones ; pharmacology ; Male ; Microdialysis ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the toxicity of Pulsatilla chinensis (Bunge) Regel saponins (PRS) against Oncomelania hupensis (O. hupensis).

METHODSO. hupensis snails were exposed to 40% and 80% of 24 h LC50 of PRS for 24 h, and then choline esterase (CHE), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activities in cephalopodium and liver of snails were determined. Niclosamide (NIC) was used as the reference molluscicide. Zebra fish lethality test was evaluated to non-target aquatic species of PRS.

RESULTSThe molluscicidal activity of PRS (LC50 at 24 h: 0.48 mg/L) was similar to that of NIC (LC50 at 24 h: 0.16 mg/L). Significant alterations about CHE, ALP, and ALT activities both in the cephalopodium and the liver of snails were observed when O. hupensis was exposed to 40% and 80% LC50 of PRS or NIC for 24 h. PRS and NIC could not affect LDH activity in the cephalopodium and the liver. Lower toxicity to fish of PRS was observed up to the highest concentration tested than NIC.

CONCLUSIONPRS, as compared with the reference molluscicide NIC, is thought to be used for the control of harmful vector snails safely.

Animals ; Molluscacides ; pharmacology ; Pulsatilla ; chemistry ; Saponins ; pharmacology ; Snails ; drug effects

OBJECTIVETo explore the relationship between the proliferation, differentiation of rat hepatic stem like cell line WB-F344 and cytokines in vitro.

METHODS(3)H thymidine labelling of new synthesized DNA was used to examine the mitogenic responsiveness of WB-F344 cells to cytokines, western blot was used to study the expression of cytokines receptors on hepatic stem cells, and apoptotic cells were detected by Flow cytometry.

RESULTSWB-F344 cells showed a proliferative response to the cytokines of hepatocyte growth factor (HGF), epidermal growth factor (EGF), fibroblast growth factor (FGF), Insulin at the dose of 80 ng/ml, and the relative cpm values are 982.95, 906.32, 863.98 and 968.67 respectively, while non response to interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) at the same dose, and an inhibition or apoptosis response to transforming growth factor-beta (TGF-beta) at 80 ng/ml with a 26.89% apoptotic rate. Western blot showed that there were HGF, EGF, FGF, TGF-beta receptors expressed on WB-F344 cells. When WB-F344 cells were cultured in the differential system (DMEM, 10% Fcs, HGF 10 to approximately 50 ng/ml, EGF 20 ng/ml, Insulin 1 microg/ml, Dex 1 micromol/L), the cells could differentiated into hepatocytes. In addition, HGF could scattered WB-F344 cells.

CONCLUSIONThe proliferation and differentiation of liver stem cells are regulated by various cytokines which may play an important role when liver is damaged seriously.

Animals ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; Cytokines ; pharmacology ; Epidermal Growth Factor ; pharmacology ; Hepatocyte Growth Factor ; pharmacology ; Insulin ; pharmacology ; Liver ; cytology ; Rats ; Rats, Inbred F344 ; Stem Cells ; cytology ; drug effects ; Transforming Growth Factor beta ; pharmacology

The objective of this study was to evaluate the role of reticulated platelets (RF) and platelet-associated antibody (PAIg) in differential diagnosis of idiopathic thrombocytopenic purpura (ITP). The immature platelet fraction (IPF) of 34 ITP patients, 36 non-ITP patients and 411 normal people were detected by Sysmex XE-5000 automated blood cell analyser and PAIgG content was detected by enzyme-linked immunosorbent assay. All results were analysed statistically. The results showed that IPF of ITP patients was significantly higher than that of non-ITP patients and normal people respectively (p < 0.05). For diagnosis of ITP, IPF positive rate of ITP patients was significantly higher than that of non-ITP patients (p < 0.05). There was no significant difference between the positive rate of PAIgG in ITP and non-ITP patients (p > 0.05). The sensibility, specificity and negative predictive value of IPF were higher than those of PAIgG, while the positive predictive value of IPF was a little lower than that of PAIgG. The area under receiver operating characteristic curve of IPF was larger than that of PAIgG. It is concluded that both IPF and PAIgG detection are valuable in differential diagnosis of ITP. The diagnostic value of IPF is better than that of PAIgG.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blood Platelets ; immunology ; Case-Control Studies ; Diagnosis, Differential ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; blood ; diagnosis ; Young Adult