1.Research on bone marrow mesenchymal stem cells in the treatment of severe acute pancreatitis
Yuqiang SHAN ; Liang LUO ; Qi XIE ; Yi WU ; Zhong JIA ; Huicheng JIN ; Guanghua FENG
Chinese Journal of Hepatobiliary Surgery 2013;19(9):711-715
Objective To investigate the mechanism of acute inflammatory response and tissue repair when rats accepted transplanted bone marrow mesenchymal stem cells (MSC) in severe acute pancreatitis (SAP).Methods A total of 40 rats were randomly divided into 4 groups which included the normal group (n=10),the model severe acute pancreatitis group (n=10),the transplanted bone marrow mesenchymal stem cells group (n =10),and the combination of bone marrow mesenchymal stem cells and granulocyte colony-stimulating factor (G-CSF) group (n=10).To cure the acute severe pancreatic injury caused by SAP,rats were injected with EDU-labeled MSCs and granulocyte colonystimulating factor (Gr-CSF).The conversion rate of MSCs to pancreatic cells or MSCs to endothelial cells was detected to assess the role of MSCs in tissue regeneration and repair.The expression of amylase,interleukin-6 (IL-6),and interleukin-10 (IL-10) in serum was detected to assess the role of MSCs in an acute inflammatory response.Results The concentrations of amylase and IL-6 were reduced and the concentration of IL-10 was increased in MSC and MSC+G-CSF groups after the onset of SAP.Flow cytometry showed a small amount of MSCs converting to endothelial or pancreatic cells,but the conversion rate was relatively low.Conclusions MSCs can play an important role in the antipre-release of inflammatory mediators,reducing acute immune response to control the acute inflammatory response in SAP.Moreover,MSCs can repair a damaged pancreas by converting into both pancreatic and endothelial cells.
2.The efficiency and function detection of NK cell differentiation from human umbilical cord hematopoietic stem cells in vitro
Qi LUO ; Jie YIN ; Yang LI ; Shan HUANG ; Xi WANG ; Jinghua HE
Tianjin Medical Journal 2015;(3):225-228
Objective To detect the efficiency and function of NK cell differentiation from human umbilical cord he?matopoietic stem cells (HSCs) in vitro. Methods CD34+hematopoietic stem cells were isolated from human umbilical cord blood, and inoculated into SCGM medium containing 20 μg/L FMS like tyrosine kinase 3 ligand (Flt-3L), stem cell fac?tor (SCF), interleukin (IL)-7, IL-15 and IL-21. And CD34+HSCs were differentiated into NK cells in directional inducing. The growth state of cells was observed. The expressions of CD56, NKG2D, NKp46, CD3, CD19 and CD34 were detected by flow cytometry in the differentiation of 7, 14, 21 and 28 d. In the differentiation of 21 d and 28 d, the differentiation cells were used as effector cells, and K562 cells as target cells. The ratios of effector cells and target cells were 8∶1, 4∶1, 2∶1 and 1∶1. The killing activity of the differentiated cells was detected by lactate dehydrogenase (LDH) cell toxicity assay and 7AAD/CFSE labeling method. Results CD34+HSCs derived from human umbilical cord blood can proliferate in vitro under appropriate condition. There were no significant differences in the expression of CD3 and CD19 between different differentia?tion stages (7, 14, 21 and 28 d, P>0.05). The expressions of CD56, NKG2D and NKp46 were significantly different (P<0.05), and the ultimate expression amount was (72.57±1.60)%, (32.83±1.29)%and (29.53±2.40)%. The expression of CD34 decreased gradually, and the lowest was (12.13 ± 2.01)%. The maximum killing activity detected by LDH cell toxicity assay and 7AAD/CFSE labeling method reached(49.91±2.76)%and (40.87±1.12)%.The killing activity of NK cells was decreased in the order of 8∶1, 4∶1, 2∶1 and 1∶1 groups (P<0.05). There was no significant difference in the killing activity between NK cells of 28 d and 21 d. Conclusion Human umbilical cord hematopoietic stem cells can differentiate into NK cells un? der appropriate conditions in vitro, and the NK cells induced from differentiation are with killing activity.
3.Cloning, Sequence Analysis and Expression of dhaT Gene from Citrobacter freundii and Purification and Property of Corresponding Recombinant Enzyme
Xiang-Hui QI ; Zhao-Fei LUO ; Yu-Tuo WEI ; Fa-Zhong CHEN ; Shan-Shan WANG ; Shou-Hai HOU ; Dong-Qing LIAO ; Ri-Bo HUANG ;
China Biotechnology 2006;0(07):-
1,3-propanediol (1,3-PD) is an important material for chemical industry, therefore, there is much interest in the production of 1,3-PD. The gene dhaT encoding 1, 3-propanediol dehydrogenase ( PDOR) of Citrobacter freundii was amplified by PCR. Sequence analysis of the similarity at the nucleotide and amino acid level between the gene encoding C. freundii PDOR and that of C. freundii ( U09771 ) were 78% and 90% , respectively. The recombinant plasmid pSE-dhaT was constructed by inserting dhaT gene into expression vector pSE380 and then transformed E. coli JM109. The recombinant strain was induced by IPTG to express dhaT. Further more the recombinant enzyme was purifed from recombinant E. coli by Ni-nitrilotriacetate affinity chromatography followed by Sephacral S-300 gel filtration. A single obvious protein about 42kDa could be obtained by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis of recombinant enzyme. The purified enzyme was used to determined enzyme property on the substrate of propionaldehyde and 1, 3-PD. The optimal temperature and optimal pH of the purified enzyme were 37℃, 8.0 for reduction and 25℃, 10. 5 for oxidation, respectively; and the kinetic property of PDOR about Km and V max were 10. 05mmol/L, 37. 27?mol/min/mg for propionaldehyde and 1. 28mmol/L, 25. 55?mol/min/mg for 1,3-PD, respectively; The deduced dhaT gene product (388 amino acids) showed a specific reduction activity of 49. 50U/mg and oxidation activity of 79. 92U/ mg. There also have a putative iron-binding motif ( G-XX-H-X-X-A-H-X-X-G-X-X-X-X-X-P-H-G) as a fingerprint pattern in the recombinant enzyme, the motif is fully conserved among these 1, 3-propanediol dehydrogenase. It is beneficial to the researches of high producing 1, 3-propanediol by gene engineering strain.
4.Protection of Tianshen Yizhi Recipe against low expression of nicotinic receptor and neurotoxicity induced by beta-amyloid peptide
Ran GU ; Ruyu LIU ; Lanjiang ZHANG ; Xiaoyan HAO ; Yan XIAO ; Xiaolan QI ; Keren SHAN ; Xilin REN ; Jun LUO ; Zhizhong GUAN
Journal of Integrative Medicine 2007;5(5):564-9
OBJECTIVE: To investigate the inhibition effects of Tianshen Yizhi Recipe (TSYZR), a compound traditional Chinese herbal medicine, on decreased expression of nicotinic acetylcholine receptor (nAChR) and the neurotoxicity as well as lipid peroxidation induced by beta-amyloid peptide (Abeta) in human SH-SY5Y neuroblastoma cells. METHODS: The SH-SY5Y cells were treated by a certain concentration of TSYZR, and then exposed to Abeta(25-35). Methyl thiazolyl tetrazolium reduction assay was carried out to understand the influences of the drugs on cellular viability. Expressions of nAChR subunits (alpha3 and alpha7) at protein and mRNA levels were detected by Western-blotting and reverse transcription polymerase chain reaction, respectively. Lipid peroxidation was measured by thiobarbituric acid to observe the capacity of antioxidant of the drugs. RESULTS: TSYZR at a safe concentration could increase alpha7 protein in the cells, inhibit decreased expressions of alpha3 and alpha7 nAChR subunit proteins, prevent lower expression of alpha7 mRNA in SH-SY5Y cells induced by Abeta, reduce the neurotoxicity and lipid peroxidation resulting from Abeta, but had no significant effect on the lower expression of alpha3 mRNA. CONCLUSIONS: TSYZR can up-regulate the expression of alpha7 nAChR subunit protein and prevent decreased expressions of nAChRs and neurotoxicity as well as lipid peroxidation induced by Abeta. This drug may play an important therapeutic role in treatment of Alzheimer disease.
5. Interaction of MicroRNAs and Endoplasmic Reticulum StressSignaling Pathways
Qi-Qi LUO ; Guang-Jin QU ; Shan-Shun LUO
Chinese Journal of Biochemistry and Molecular Biology 2022;38(2):191-196
Endoplasmic reticulum stress (ERS) is a cellular defensive response to restore homeostasisand reduce the protein load. Over-activation of ERS can induce cell differentiation, proliferation, apoptosis, and autophagy. MicroRNAs (miRNAs) are endogenous non-coding RNAs (ncRNAs) thatregulate the expression of key proteins and genes in the ERS signaling pathway through post-transcriptional action. Meanwhile, activated ERS signaling pathway can indirectly regulate the expressionand function of target genes by decreasing miRNA stability. Based on a brief introduction of ERS classicalsignaling pathways, this paper further elaborated how microRNAs regulate ERS signaling pathways topromote apoptosis and proliferation, and what effect they would have on the expression profile of diseasesbased on this association. We also summarize the regulation of ERS on miRNAs expression and thecurrent research status. The mutual regulation between the two could provide a new idea for the follow-upresearch on the therapeutic targets of diseases.
6.The status on care and nutrition of 774 children staying in rural areas while parents were in towns
Xi-Chun PENG ; Jia-You LUO ; Kuan-Bao YAO ; Ru-Shan HU ; Qi-Yun DU ; Ming-Yuan ZHU
Chinese Journal of Epidemiology 2008;29(9):860-864
Objective To understand the status on care and nutrition of children living in the rural areas (so called 'left-behind' children) while their parents were seeking for jobs in the urban areas. Methods Cross-sectional study was employed in this investigation.The group of‘left-behind' children (n=774) and the comparison group (n=774) were identified.The ascertainment methods mainly included questionnaire,anthropometries measurements,food-frequency,and laboratory examination.Results be poorly attended (X2=6.671,P=0.036;X2=15.053,P=0.001 ).Mothers who chose to work outside of their households would tend to choose bottle-feeding or decrease the duration of breastfeeding for their infants (X2=5.051,P=0.031;t=-7.201,P=0.001).The intake of milk and bean products in children were obviously lower than that seen in the control group (t=-2.150,P=0.032 ;t=-2.054,significant difference when comparing with the control group (X2=4.560,P=0.033 ).Conclusion communities,parents and extended families would facilitate more attention and effective intervention programs to improve the situation.
7.A case-control study on genetic and environmental factors regarding polydactyly and syndactyly
Jia-You LUO ; Chu-Hui FU ; Kuan-Bao YAO ; Ru-Shan HU ; Qi-Yun DU ; Zhi-Yu LIU
Chinese Journal of Epidemiology 2009;30(9):903-906
king condition before pregnancy were associated with the development of polydactyly and syndactyly.
8.Clinical Analysis of Mitoxantrone Liposome in the Treatment of Children with High-Risk Acute Myeloid Leukemia
Yu-Qing JIAO ; Hao XIONG ; Zhi CHEN ; Li YANG ; Fang TAO ; Ming SUN ; Shan-Shan QI ; Wen-Jie LU ; Zhuo WANG ; Yu DU ; Lin-Lin LUO
Journal of Experimental Hematology 2024;32(2):365-369
Objective:To investigate the safety and efficacy of mitoxantrone liposome in the treatment of children with high-risk acute myeloid leukemia(AML).Methods:The children with high-risk AML who received the mitoxantrone liposome regimen at Wuhan Children's Hospital from January 2022 to February 2023 were collected as the observation group,and the children with high-risk AML who received idarubicin regimen were enrolled as controls,and their clinical data were analyzed.Time to bone marrow recovery,the complete remission rate of bone marrow cytology,the clearance rate of minimal residual disease,and treatment-related adverse reactions were compared between the two groups.Results:The patients treated with mitoxantrone liposome showed shorter time to recovery of leukocytes(17 vs 21 day),granulocytes(18 vs 24 day),platelets(17 vs 24 day),and hemoglobin(20 vs 26 day)compared with those treated with idarubicin,there were statistical differences(P<0.05).The effective rate and MRD turning negative rate in the observation group were 90.9%and 72.7%,respectively,while those in the control group were 94.1%and 76.4%,with no statistical difference(P>0.05).The overall response rate of the two groups of patients was similar.Conclusion:The efficacy of mitoxantrone liposome is not inferior to that of idarubicin in children with high-risk AML,but mitoxantrone liposome allows a significantly shorter duration of bone marrow suppression and the safety is better.
9.Clinical Analysis of Reversible Posterior Encephalopathy Syndrome after Allogeneic Hematopoietic Stem Cell Transplantation in Children
Zuo-Feng LI ; Hao XIONG ; Zhi CHEN ; Li YANG ; Ming SUN ; Wen-Jie LU ; Shan-Shan QI ; Fang TAO ; Lin-Lin LUO ; Yu-Qing JIAO
Journal of Experimental Hematology 2024;32(5):1560-1565
Objective:To summarize the clinical features of reversible posterior encephalopathy syndrome(PRES)after allogeneic hematopoietic stem cell transplantation(allo-HSCT)in children.Methods:The clinical data of six children who developed PRES after undergoing allo-HSCT in the Department of Hematology of Wuhan Children's Hospital from June 2016 to December 2022 were retrospectively analyzed,and their clinical characteristics,imaging examination,laboratory examination,and treatment regression were summarized.Results:Among 281 children underwent allo-HSCT,6 cases(2.14%)developed PRES,with a median age of 5.1(1.5-9.7)years old.4 cases underwent related haploidentical donor transplantation,and 2 cases underwent sibling allografting and unrelated donor allografting donor transplantation,respectively.All six children had an acute onset of illness,with clinical manifestations of nausea and vomiting,seizures,psychiatric disorders,visual disturbances.The five cases elevated blood pressure.All children with PRES were treated with oral immunosuppressive drugs during seizures,and 3 cases were combined with different degrees of graft-versus-host disease.Most of the children showed effective improvement in clinical symptoms and imaging after adjusting/discontinuing suspected medications(cyclosporine,etc.)and symptomatic supportive treatments(oral antihypertensive,diazepam for antispasmodic,mannitol to lower cranial blood pressure),and one of them relapsed more than 8 months after the first seizure.Conclusion:PRES is rare after hematopoietic stem cell transplantation in children,and its onset may be related to hypertension,cytotoxic drugs,graft-versus-host disease,etc.Most of them can be recovered after active treatment,but not completely reversible,and the prognosis of those who combined with TMA is poor.
10.Therapeutic effects and complications of percutaneous pedicle screw fixation for thoracolumbar fractures.
Peng LUO ; Liang-Feng XU ; Wen-Fei NI ; Xiang-Yang WANG ; Yan LIN ; Fang-Min MAO ; Qi-Shan HUANG ; Hua-Zi XU ; Yong-Long CHI
Chinese Journal of Surgery 2011;49(2):130-134
OBJECTIVETo investigate the therapeutic effects and complications of percutaneous pedicle screw fixation for thoracolumbar fractures.
METHODSFrom January 2002 to December 2008, 103 patients with thoracolumbar fractures were treated with percutaneous pedicle screw fixation, including 75 males and 28 females, the average age was 45.6 years (range, 18 - 72 years). All of them were of no neurological deficits. There were 65 cases of traffic injury, 23 cases of fall injury and 15 cases of smashed injury. According to the Denis classification, 64 patients were of compression fractures, and 39 patients of burst fractures. There were 5 cases had fractures in T(11), 30 in T(12), 42 in L(1), 15 in L(2), 4 in L(3), 3 in L(4), 2 in T(11-12), 1 in L(1-2), and 1 in L(2-3). Radiological examinations, including X-ray and CT examinations, and clinical examinations were carried out to evaluate the therapeutic effects.
RESULTSTwenty one patients were lost to follow up, the remaining were followed up from 10 to 48 months with an average of 27.4 months. Before the operation, the vertebral height, the kyphosis angle and the occupation of spinal canal were (54.5 ± 8.7)%, 16.4° ± 2.9° and 1.2 ± 1.0, and were improved to (88.6 ± 6.4)%, 11.6° ± 2.7° and 0.5 ± 0.6 respectively after the operation. Preoperatively the visual analogue scale and the Oswestry disability index were 8.0 ± 1.2 and 41.2 ± 9.3, and were improved to 1.7 ± 1.8 and 6.7 ± 5.6 postoperatively, respectively. All of these values between pre- and post-operatively were significantly different (P < 0.01). Screw misplacement was found in 7 patients, superficial wound infection in 1, screw breakage in 3, screw dislodgment in 2, cement leakage in 5, transient neurological symptoms in 4, and 8 patients with low back pain remained, of which 2 patients required occasional oral analgesics. Bone fusion achieved in all cases.
CONCLUSIONSThe clinical efficacy of percutaneous pedicle screw fixation is similar with conventional open surgery. With the advantages of convenient procedure, less invasive, and rapid recovery, percutaneous pedicle screw fixation is an alternative method for thoracolumbar fractures without neurological deficits.
Adolescent ; Adult ; Aged ; Bone Screws ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; Treatment Outcome ; Young Adult