Induced pluripotent stem cells can differentiate into a variety of cell types,which promote the development of human disease model,drug toxicity screening and sources of autologous cells.However,there have been many problems in the induced pluripotent stem cells reprogramming,such as safety and low efficiency.Small molecules are considered as a promising method to improve the reprogramming processes of induced pluripotent stem cell,and more and more small molecules have been identified to maintain stem cell self-renewal,providing a new approach to produce the desired reprogramming cells.

BACKGROUND:Astragaloside Ⅳ is a major component of Huangqi,promoting proliferation and differentiation of bone marrow mesenchymal stem cells;however,the mechanism has been less reported yet.OBJECTIVE:To explore the effect of Astragaloside Ⅳ on expression of multiple hematopoietic growth factors in bone marrow mesenchymal stem cells.METHODS:Bone marrow mesenchymal stem cells were isolated from adult Wistar rats by using the method of adhesive culture and clone,and they were then plated on 96-well plate and separately incubated with 100 uL Astragaloside Ⅳ(25,50,100,200 g/L)for 72 hours.The cells in the control group were cultured with an equal volume of DMEM-LG culture liuquid.Indirect immunofluorescence was used to detect the biological activity,MTT method was used to evaluate the effect of Astragaloside Ⅳ on proliferation and differentiation of bone marrow mesenchymal stem cells,and RT-PCR method was used to measure the expression of hematopoietic growth factors in bone marrow mesenchymal stem cells.RESULTS AND CONCLUSION:The 3~(rd)-passage bone marrow mesenchymal stem cells highly expressed CD44 but lowly expressed CD45.As compared with control group,Astragaloside Ⅳ promoted proliferation of bone marrow mesenchymal stem cells in a time/dosage-dependent manner,in particular,the 200 g/L Astragaloside IV and 72-hour intervention(P＜ 0.05).SCF expression was significantly increased in the drug group compared with control group(P ＜ 0.01);however,TPO,GM-CSF,and TGF-β1 expressions were not changed significantly(P ＞ 0.05).Moreover,interleukin-3 expression was not found in the bone marrow mesenchymal stem cells.Astragaloside Ⅳ promoted in vitro proliferation of bone marrow mesenchymal stem cells,possibly involving in SCF secretion.

ObjectiveTo analyze the Uridine diphosphoglucuronyl transferase 1 A1 gene (UGT1A1) mutation in non-he-molytic indirect hyperbilirubinemia.MethodsA female patient was diagnosed with type I Crigler-Najjar syndrome (CNS-I) after excluding hemolysis and hypothyroidism. Phototherapy treatment is effective while oral phenobarbital was not. UGT1A1 were ampliifed by polymerase chain reaction and DNA was sequenced.ResultsThe patient was compound heterozygote of c.1070A>G p. (Gln357Arg) and 1091C>T p. (pro364Leu) and diagnosed with CNS-I. Heterozygotes of the mutation were found in her parents.ConclusionsIn patients highly suspected of CNS, the genetic tests should be carried out as soon as possible.

Objective To analyze the gene abnormality and liver pathology in Shwachman-Diamond syndrome (SDS) in a child. Method The clinical data of one child with SDS were analyzed retrospectively. Result A male patient was 1 month old at onset with neutrophil decrease as the first manifestation, accompanied by anemia, elevated transaminase and repeated infection. Exocrine pancreatic dysfunction was atypical. Pathological examination of liver biopsy showed slight damage of liver cells under light microscope. The blood samples of child and parents were collected, and homozygous mutations of SBDS (NM_016038.2) Intron2 c.258+2T>C p.? were detected by two generation gene sequencing. And these mutations were from both his parents. Conclusion Gene testing is helpful in diagnosing SDS and we suggest that liver biopsy should be performed if condition allows.

Objective To study the inhibitory effects of dehydroepiandrosterone (DHEA) and its metabolites-dehydroepiandrosterone sulfate (DHEAs) on the proliferation of HepG2 and HT-29 and their mechanism.Methods HepG2 and HT-29 were incubated by DHEA or DHEAs with different concentrations (1,10,50,100 and 200 ?mol?L-1) for 8,24,48,72 h and routine culture was used as control.The inhibitory rate was detected by using MTT chromometry and BrdU assay respectively.The activities of 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGR),glucose -6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) were examined simultaneously.Results ①MTT chromometry:DHEA with different concentrations obviously inhibited the growth of HepG2 and HT-29 cells compared with control group(P0.05).②BrdU assay:the growth of cells were significantly inhibited by DHEA with concentrations of 50,100 and 200 ?mol?L-1,especially to HepG2 cells(P0.05).Conclusion DHEA has strong anti-proliferative effects on both HepG2 and HT-29 cell lines and inhibitory effects on the activities of G6PD or HMGR,however,DHEAs has no obvious effect.

ObjectiveTo study the role and mechanism of resisitin in prophylactic and therapeutic treatments of rosiglitazone,a specific peroxisome proliferator-activated receptor-γ(PPARγ) ligand,in rats with severe acute pancreatitis (SAP) and pancreatitis-associated pulmonary injury.MethodsThe levels of amylase ( AMY ),Resistin,TNF-α,IL-1 β and C reactive protein (CRP) in blood plasma,lung myeloperoxidase ( MPO ) activity,pancreas/body weight ratio and lung wet/dry weight ratio were evaluated.Pancreatic and pulmonary pathology were observed.The expression of resistin in pancreas was detected byimmunohistochemistry.The gene expression of resistin mRNA was investigated by real-time PCR.ResultsBoth prophylactic and therapeutic treatments with rosiglitazone could obviously ameliorate the levels of AMY,resistin,TNF-αt,IL-1β and CRP ( all P ＜ 0.01 ).Compared with the control group,both prophylactic and therapeutic treatment groups were higher( all P ＜ 0.01 ).The prophylactic treatment group was not different from the therapeutic treatment group.Both prophylactic and therapeutic treatments with rosiglitazone could significantly reduce pancreas/body weight ratio,pancreatic pathology,MPO,pulmonary pathology ( all P ＜ 0.01 ).Compared with the SAP group,the expression of resistin mRNA in the prophylactic and therapeutic treatment groups were obviously decreased.ConclusionRosiglitazone could obviously ameliorate pancreatitis and pulmonary injury induced by L-arginine.

Clinical data of 108 patients with adult-onset Still's disease(AOSD)were analyzed.The results showed that 56 patients(51.9%)developed liver damage and abnormal liver functioil(alanine aminortransferase and aspartate aminortransferase were significantly increased).Gender,age,time from disease onset to confirmed diagnosis,fever,rash,arthritis,pharyngeal pain,splenomegaly,lymphadenopathy,anemia,leukocvtosis,thrmbocytosis,and increased erythrocyte sedimentation rate,serum C-reactive protein,and serum iton protein showed no significant difierence between the two groups (P＞0.05).Clinical symptoms of liver damage were atypical.Among the 56 patients with liver damage,8 were presented with gastrointestinal symptoms,and 24 showed enlarged liver iu B.ultrasound examination.Our results suggest the prevalence of liver damage in AOSD patients may be hiSh,which could attribute to the disease itself and the medication.The patients would expect good response to corticosteroid treatment and satisfactory outcomes.

Objective To investigate the clinical value of serum alpha fetoprotein ( AFP) combined with abnormal prothrombin(PIVKA-Ⅱ) in the detection of primary liver cancer.Methods From May 2017 to October 2017,80 cases of primary liver cancer in Department of Hepatobiliary Surgery of Deyang People 's Hospital,100 cases of non HCC patients ( including chronic liver disease ,cirrhosis,metastatic liver cancer ) were selected,and another 100 healthy people were selected as control group.The levels of serum AFP and PIVKA -Ⅱ were detected by SIEMENS XP automatic immunoassay analyzer and lumipulseg 1200 automatic immune analyzer respectively.All data were analyzed by SPSS17.0 analysis software.Results Comparison between primary liver cancer and non -primary liver cancer:the levels of serum AFP and PIVKA -Ⅱ(F＝1.763;F＝1.788,sig＝0.000,P＜0.05).Comparison between non-primary liver cancer and normal group:the levels of serum AFP and PIVKA -Ⅱ(F＝0.896;F＝0.346,sig＝0.000,P＜0.05).Serum AFP had a sensitivity of 75.0%and a specificity of 71.5%in the diagnosis of primary liver cancer, while PIVKA -Ⅱ had a sensitivity of 82.0% and a specificity of 78.7%.The diagnosis sensitivity and specificity of AFP combined with PIVKA -Ⅱreached to 92.0% and 95.0%.The sensitivity of AFP combined with PIVKA-Ⅱin the diagnosis of primary liver cancer had statistically significant differences compared with AFP and PIVKA-Ⅱalone(χ2＝4.112,P＝0.008;χ2＝2.991,P＝0.010).Conclusion Combined detection of serum AFP and PIVKA-II has important clinical significance in the diagnosis of primary liver cancer .

OBJECTIVE: To explore the genetic basis for a child with infantile liver failure syndrome type 2 (ILFS type 2). METHODS: Clinical features of the child were analyzed. Next generation sequencing was also carried out for him. RESULTS: The child was found to harbor compound heterozygous variants of the NBAS gene, which included a novel nonsense c.2746A>T (p.R916X, 1456) variant in exon 24 and a missense c.3596G>A (p.C1199Y) mutation in exon 31, which has been associated with ILFS type 2. The two variants were respectively inherited from his father and mother. CONCLUSION The compound heterozygous variants of c.3596G>A and c.2746A>T of the NBAS gene probably underlay the ILFS type 2 in this child.
Child ; Exons/genetics* ; Genetic Testing ; High-Throughput Nucleotide Sequencing ; Humans ; Liver Failure ; Male ; Mutation

Objective To evaluate the effect of transcutaneous neuromuscular electrical stimulation on prevention of intensive care unit-acquired weakness (ICU-AW) in chronic obstructive pulmonary disease (COPD) patients with mechanical ventilation. Methods A prospective randomized controlled study was conducted. Sixty COPD patients aged 18-85 years old who were accepted mechanical ventilation therapy admitted to general intensive care unit (ICU) of the First Affiliated Hospital of Hunan University of Medicine from October 2017 to October 2018 were enrolled. Patients were divided into control group (n = 30) and intervention group (n = 30) by random number table method. All patients were accepted routine treatment, and on this basis, the intervention group was applied transcutaneous neuromuscular electrical stimulation on the extremities (twice a day, 30 minutes each time) after 24 hours of admission until ICU discharge. The Medical Research Council muscle strength score (MRC-Score), grip strength, incidence of ICU-AW on the 7th day after admission and on the day of ICU discharge; modified Barthel index score on the day of ICU discharge; and duration of mechanical ventilation, the length of ICU stay, and the length of hospital stay were compared between the two groups. Results Twenty-nine and 27 patients in the control group and the intervention group respectively finally completed the study in dividually. There was no significant difference in gender, age, Barthel index score before 2 weeks of ICU admission, body mass index or acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) in ICU between the two groups. There was no significant difference in the MRC-Score, grip strength or incidence of ICU-AW on the 7th day after ICU admission between the two groups. Compared to the control group, the MRC-Score, grip strength and Barthel index score in the intervention group were significantly increased [MRC-Score: 55.97±8.43 vs. 46.32±7.36, grip strength (kg): 33.46±11.62 vs. 27.42±9.64, Barthel index score:46.04±5.46 vs. 42.13±3.32, all P < 0.05], the incidence rate of ICU-AW was significantly decreased [7.4% (2/27) vs. 31.0% (9/29), P < 0.05], and duration of mechanical ventilation, the length of ICU stay, the length of hospital stay were significantly shortened [duration of mechanical ventilation (days): 5.12±2.01 vs. 7.24±4.35, the length of ICU stay (days): 8.34±2.36 vs. 10.45±2.62, the length of hospital stay (days): 13.21±2.21 vs. 15.38±3.67, all P < 0.05]. Conclusion Transcutaneous neuromuscular electrical stimulation can effectively improve the muscle strength of COPD patients with mechanical ventilation and reduce the incidence of ICU-AW.