AIM: Mesenchymal stem cells (MSCs) isolated from human umbilical cord blood can differentiate into hepatocyte-like cells under a suitable induction condition. The optimal condition is still unclear. This study investigated the feasibility of differentiation of MSCs isolated from human umbilical cord blood into hepatocyte-like cells with hepatocyte growth factor (HGF) and interleukin-6 (IL-6) in vitro to find a new cell source for live tissue engineering. METHODS: The experiment was performed from March to September 2007 at the Central Laboratory of First Hospital of Lanzhou University. ①Three cases of umbilical cord blood were collected from full-term pregnant women. Pregnant women had signed an informed consent. The experiment was approved by Hospital’s Ethical Committee. ②MSCs from umbilical cord blood were separated by density gradient centrifugation and adherent method. Cell surface molecule was measured by flow cytometry. Third passage of cells were divided into four groups and cultured in DMEM medium with HGF (HGF group), IL-6 (IL-6 group), HGF+IL-6 (HGF+IL-6 group) or no growth factor (control group). ③The characteristics of proliferation and growth of MSCs were studied by microscopy and methyl thiazolyl tetrazolium (MTT). The phenotypes of MSCs were identified by flow cytometry and immunohistochemical method. Albumin levels in culture supernatants were determined with enzyme-labeled immunosorbent assay (ELISA). RESULTS: ①Growth and division of adherent cells obtained from human umbilical cord blood were good. The shape of MSCs changed into triangle, polygonal or round on days 21-28 in induction groups. ②Positive staining reaction for alpha fetoprotein (AFP) on day 7, for CK18 on days 21, 28 after induction. Albumin production by induced MSCs increased in a time-dependent manner. The positive percents of every hepatocytic marker were higher in the HGF+IL-6 group than in the IL-6 group and HGF group (P

Objective To explore the possibility of differentiation of mesenchymal stem cells isolated from patients with hepatocirrhosis into hepatocyte-like cells induced by EGF and HGF and to lay basis for transplanted autologous bone marrow MSCs in treatment of liver disease at terminal stage.Methods Bone marrow cells were obtained from volunteers with liver cirrhosis.MSCs were separated by density gradient centrifugation and were cultured through adhere culture.MSCs were cultured in DMEM medium with HGF,EGF,HGF+EGF or no growth factor.The phenotypes of MSCs were identified by flow cytometry,immunohistochemistry,and Albumin levels in culture supernatants were determined by ELISA.Results Growth and division of adherent cells obtained from the patients with hepatocirrhosis were good and the phenotypes of MSCs were CD29 positive and CD34 negative.The shape of MSCs changed from long fusiform to polygonal or round on 21th-28th days in grow factor induced groups.Immunocytochemical analysis for CK18 and AFP showed positive staining reaction for AFP on 7th day,for CK18 on 21st and 28th day in grow factor induced groups with MSCs-induced Alb production increasing in a time-dependent manner.No markers of hepatocyte linear cells were detected in no growth factor induction group.Conclusion Both HGF and EGF can induce mesenchymal stem cells to differentiate hepatocyte-like cells alone or coordinately.

BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into hepatic stem cells in a specific environment, and participate in the repair and reconstruction of liver function.OBJECTIVE:To investigate the therapeutic effect of autologous bone marrow mesenchymal stem cell transplantation on decompensated hepatitis B.METHODS:Eighty-four patients with decompensated hepatitis B were randomly divided into two groups:normal group (n=42) with symptomatic treatment, and oral entecavir, Fuzheng Huayu Capsule; stem cell group (n=42) with the left and right hepatic artery transplantation of autologous bone marrow mesenchymal stem cells (1×106/kg) based on conventional treatments. Degree of liver fibrosis, liver function and score on Model for End-Stage Liver Disease (MELD) system, 1-year survival rate of patients were detected and analyzed with statistics before and 12, 24 weeks after treatment.RESULTS AND CONCLUSION:(1) Liver fibrosis after treatment in two groups:hyaluronic acid, laminin, type Ⅲ collagen and type IV collagen levels after treatment were lower than those before treatment in both two groups (P < 0.05), while these indexes in the stem cell group were lower than those in the normal group at 12 and 24 hours after treatment (P <0.05). (2) Liver function:after treatment, decreased alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels were found in both two groups (P < 0.05), and albumin, cholinesterase, prothrombin activity levels were higher than those before treatment (P < 0.05). The alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels in the stem cell group were lower than those in the normal group at 12 and 24 weeks after treatment, while the cholinesterase level was higher in the stem cell group (P < 0.05). (3) MELD scores:MELD scores were both decreased in the two groups after treatment (P < 0.05), and lower in the stem cell group compared with the normal group at 24 weeks after treatment (P < 0.05). (4) The 1-year survival rate was higher in the stem cell group (69%) than the normal group (50%; P < 0.05). To conclude, the use of autologous bone marrow mesenchymal stem cell transplantation in the treatment of hepatitis B can significantly improve the patients' liver fibrosis and liver function, and improve the 1-year survival rate of patients.

Objective To investigate the effect of stem cells transplantation on immune function,liver function and related indexes in the patients with end-stage liver disease(ESLD).Methods A total of 163 cases of ESLD in Nanyang Municipal Central Hospital of Affiliated Hospital of Zhengzhou University were selected and divided into 2 groups by the randomized single blind method.Eighty-one cases in the control group were given the conventional symptomatic treatment,while 82 cases in the observation group received bone marrow mesenchymal stem cell(BMSC) transplantation based on the control group.The changes of immune function,liver function,alpha fetoprotein(AFP),rate of prothrombin activity(PTA) and plasma total protein(TP) level before and after treatment were observed in the two groups.Results The immune function indexes CD3+,CD4+,CD8+,CD4+/CD8+,TP and PTA levels after treatment in the observation group were significantly higher than those before treatment and in the control group (P＜0.05),while the levels of AFP,alanine aminotransferase (ALT),aspartate aminotransferase (AST) and total bilirubin (TBIL)were lower than those before treatment in the same group and the control group(P＜0.05).There was no statistically significant difference in complication occurrence rate between the two groups(P＞0.05).Conclusion BMSC transplantation for treating ESLD can improve the immune function,improves the liver function and reduces the AFP level.

Objective To explore the sleep characteristics of adult partial epilepsy patients. Methods 45 patients newly diagnosed as adult partial epilepsy and 40 cases of healthy controls were monitored with sleep encephalogram (EEG), the sleep parameters were analyzed.Epworth Sleepiness Scale (ESS) was used to evaluate the daytime sleepiness next day. The sleep parameters and ESS score were compared between 2 groups, and the correlation between sleep parameters and ESS score in the patients were analyzed. Results The ESS score was higher in the patients than in the controls (P<0.05); Compared with the controls, the total sleep time decreased (P<0.05), the sleep efficiency reduced (P<0.05), the percentage of non-rapid eye movement sleep (NREM) stages 1 and 2 and the wakefulness stage increased (P<0.05), the percentage of NREM stages 3 and 4 decreased (P<0.05) in the patients; There was no significant difference in total recording time, sleep latency, REM latency, and arousal times between the patients and the controls (P>0.05). The ESS score was higher in the patients who attacked 3 times a month or more (group A) than less than 3 (group B). Compared with group B, the total sleep time decreased (P<0.05), the sleep efficiency reduced (P<0.05), the percentage of NREM stages 1 and 2 and the wakefulness stage increased (P<0.05), the percentage of NREM stages 3 and 4, and REM decreased (P<0.05) in group A. There was no significant difference in sleep latency, REM latency and arousal times between 2 groups (P>0.05). The ESS score of the patients was negatively correlated with sleep efficiency (r=-0.49, P<0.05) and the percentage of NREM stages 3 and 4 (r=-0.54, P<0.05), and was not correlated with other sleep parameters (P>0.05). Conclusion The sleep architecture of patients with adult partial epilepsy is abnormal, and their daytime sleepiness is more than healthy persons. Epileptic seizure can aggravate sleep architecture abnormality and daytime sleepiness. Abnormal sleep parameters are related with ESS score.

We established a rapid detection method of New Delhi Metallo-beta-Lactamase Gene (NDM-1) based on Loop-mediated Isothermal Amplification (LAMP). With the application of LAMP, we designed four sets of LAMP premiers, using NDM-1 gene as the target sequence, and selected the set of optimal primers. Meanwhile, we established optimal reaction systems and conditions to carry out the sensitivity and specificity experiments. The experiment results showed that the whole detection process took only one hour and could be observed visually. In the experiment of sensitivity, NDM-1 gene had a detection limit of 6 copies in each reaction. In the experiment of specificity, we detected NDM-1 gene in 4 pathogen strains (Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae), and the total DNA from intestinal microbes and the total DNA from soil microbes. We had not detected the amplification reactions. The detection method established could rapidly detect NDM-1 gene and visualize the experiment result. The method is easy to operate and has high sensitivity and specificity and thus has great application value in basic research laboratories, emergent detection and spot detection.
Bacteria ; enzymology ; genetics ; Bacteriological Techniques ; methods ; DNA, Bacterial ; genetics ; Escherichia coli ; enzymology ; genetics ; Klebsiella pneumoniae ; enzymology ; genetics ; Nucleic Acid Amplification Techniques ; methods ; Sensitivity and Specificity ; Staphylococcus aureus ; enzymology ; genetics ; Streptococcus pneumoniae ; enzymology ; genetics ; beta-Lactamases ; genetics

Objective To investigate the influence of collateral circulation recruitment on cognitive functions in patients with severe unilateral stenosis or unilateral occlusion of the internal carotid artery by using TCD.Methods A total of 176 patients with severe unilateral stenosis or unilateral occlusion of the internal carotid artery were enrolled and were divided into a single vessel collateral group(n=80,45.5 ％),a multiple vessel collateral group(n=74,42.1％) and a no collateral group(n=22,12.5 ％).In order to study the influence of single vessel collateral circulation on cognitive functions,the single vessel collateral group was further divided into an anterior communicating subgroup(AcoA),a posterior communicating subgroup(PcoA),an ocular artery subgroup(OA),and a normal control subgroup.All patients and 34 normal controls(NC) received MoCA,and scores for the overall assessment and individual domains were analyzed.Results Compared with the control group (26.3± 1.1,4.7 ± 0.5,2.0 ± 0.0,3.6 ± 1.0),the multiple vessel collateral group,the single vessel collateral group and the no collateral group had lower overall scores (24.2 ± 1.7,21.9 ± 2.3,19.0 ± 2.4),lower executive/visuospatial function(3.9 ± 0.7,3.2 ± 0.8,2.4 ± 0.6),lower abstraction (1.7 ± 0.5,1.6±0.5,1.3±0.5),and lower delayed recall(2.9±0.8,1.9±0.8,1.6±0.5)(F=80.52,63.21,12,48.99,all P＜0.05);both collateral groups had lower scores in subtraction and attention (4.6±0.6 vs.5.2±0.7,4.3±0.7 vs.5.2±0.7);the no collateral group had lower scores in orientation(4.7±0.7 vs.5.7±0.5)(P＜0.05).Compared with the multiple vessel collateral group,both the single vessel collateral group and the no collateral group had lower overall scores(21.9±2.3 vs.24.2 ± 1.7,19.0± 2.4 vs.24.2 ± 1.7),executive/visuospatial function (3.2 ± 0.8 vs.3.9 ± 0.7,2.4±0.6 vs.3.9±0.7),subtraction and attention(4.6±0.6 vs.5.1±0.5,4.3±0.7 vs.5.1±0.5) (all P＜ 0.05);the no collateral group had lower scores in abstraction (1.3 ± 0.5 vs.1.7 ± 0.5),delayed recall(1.6 ± 0.5 vs.2.9 ± 0.8) and orientation (4.7 ± 0.7 vs.5.7 ± 0.5) (all P ＜ 0.05).Compared with the single vessel collateral group,the no collateral group had lower overall scores(19.0 ±2.4 vs.21.9±2.3),executive/visuospatial function(2.4±0.6 vs.3.2±0.8)and orientation(4.7± 0.7 vs.5.7 ± 0.6) (all P ＜0.05).In single vessel collateral patients,the AcoA subgroup had higher MoCAscores than the PcoA subgroup(22.9± 1.7 vs.21.2±2.7) (P＜0.05),and the AcoA subgroup had higher scores in delayed recall than the OA subgroup(2.2±0.8 vs.1.7±0.6) (P＜ 0.05).Conclusions Severe unilateral stenosis or unilateral occlusion in the internal carotid artery can result in cognitive impairment,especially in executive/visuospatial function,abstraction,delayed recall,subtraction and attention.However,collateral circulation can protect cognitive function in patients with unilateral internal carotid artery stenosis or occlusion and multiple vessel collateral circulation is more effective than single vessel collateral circulation,and AcoA is more effective than either PcoA or OA,but the difference between PcoA and OA is not significant.

OBJECTIVES: To investigate the effect of recombinant human fibroblast growth factor 21 (rhFGF21) on the proliferation and mineralization of cementoblasts and its mechanism. METHODS: Hematoxylin eosin, immunohistochemical staining, and immunofluorescence were used to detect the expression and distribution of fibroblast growth factor 21 (FGF21) in rat periodontal tissues and cementoblasts (OCCM-30), separately. Cell Counting Kit-8 was used to detect the proliferation of OCCM-30 under treatment with rhFGF21. Alkaline phosphatase staining and Alizarin Red staining were used to detect the mineralization state of OCCM-30 after 3 and 7 days of mineralization induction. The transcription and protein expression of the osteogenic-related genes Runx2 and Osterix were detected by real-time quantitative polymerase chain reaction (PCR) and Western blot analysis. The expression levels of genes of transforming growth factor β (TGFβ)/bone morphogenetic protein (BMP) signaling pathway in OCCM-30 were detected through PCR array analysis. RESULTS: FGF21 was expressed in rat periodontal tissues and OCCM-30. Although rhFGF21 had no significant effect on the proliferation of OCCM-30, treatment with 50 ng/mL rhFGF21 could promote the mineralization of OCCM-30 cells after 7 days of mineralization induction. The transcriptional levels of Runx2 and Osterix increased significantly at 3 days of mineralization induction and decreased at 5 days of mineralization induction. Western blot analysis showed that the protein expression levels of Runx2 and Osterix increased during mineralization induction. rhFGF21 up-regulated Bmpr1b protein expression in cells. CONCLUSIONS rhFGF21 can promote the mineralization ability of OCCM-30. This effect is related to the activation of the TGFβ/BMP signaling pathway.
Humans ; Rats ; Animals ; Dental Cementum ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Cell Differentiation ; Bone Morphogenetic Proteins/metabolism* ; Transforming Growth Factor beta/pharmacology*

ObjectiveTo investigate the high-risk detection rate and aggregation of cardiovascular diseases（CVD） in 8 districts of Shanghai and influencing factors， and to provide scientific references for prevention and control of CVD. MethodsBased on the Cardiovascular Disease Screening and Management Program in Shanghai from 2016 to 2021， 104 685 participants aged 35 to 75 in 8 districts of Shanghai were selected for analysis. χ² test and multivariate logistic regression were used for statistical analysis of the influencing factors of CVD and aggregation of CVD. ResultsThe proportion of high-risk CVD individuals in the population was 19.17%， including the high-risk individuals with hypertension （8.65%）， dyslipidemia （6.33%）， CVD history （5.58%）， and WHO assessed risk ≥20% types （2.69%）， respectively. Old age， overweight and obesity， central obesity， smoking， drinking， farmers， unmarried， and low family income were the risk factors of CVD， while high education level was the protective factor. In the participants， 16 323 people （81.34%） were classified as CVD high-risk groups； The number of aggregation of 1， 2 and ≥3 high risk types of CVD were 16 323（81.34%）， 3 236（16.13%）， 509（2.54%）， respectively. Old age， low education level， low annual family income， farmers， unmarried， smoking， drinking， overweight， obesity and central obesity were associated with the risk of aggregation of high risk types of CVD， and the correlation strength increased with the increase of aggregation types. ConclusionThe prevention and control of CVD in Shanghai should focus on the hypertension， elderly， overweight， obesity， central obesity， smoking， drinking， low educated， low family income， farmers and unmarried people， and targeted intervention measures should be taken to reduce the risk of CVD among residents.

Chronic heart failure (CHF) is a severe public health problem with increasing morbidity and mortality, any treatment targeting a single session is insufficient to tackle this. CHF is characterized by reduced cardiac output resulting from neurohumoral dysregulation and cardiac remodeling, which might be related to oxidative stress, inflammation, endoplasmic reticulum stress, apoptosis, autophagy, mitochondrial function, and angiogenesis. These molecular mechanisms interact with each other through crosstalk. Historically, Chinese medicinal herbs have been widely applied in the treatment of CHF, and therapeutic effects of Chinese medicinal herbs and their ingredients have been scientifically confirmed over the past decades. Traditional Chinese medicine (TCM) with multiple components can confront the different pathogenesis of CHF through multiple targets. This review analyzes commonly used TCM patent drugs and TCM decoctions that are applicable to different stages of CHF based on clinical trials. Diverse bioactive ingredients in Chinese medicinal herbs have been found to treat CHF via multiple molecular mechanisms. This review comprehensively covers the key works on the effects and underlying mechanisms of TCM, herbal ingredients and synergistic effects of constituent compatibility in treating CHF, providing additional ideas to address this threat.