Comparison of interscalene brachial plexus block and superior trunk block with ropivacaine in shoulder arthroscopic surgery / 药学实践杂志

Objective To compare the clinical effects of interscalene brachial plexus block and superior trunk block in arthroscopic shoulder surgery with 0.25% ropivacaine. Methods 46 patients undergoing shoulder arthroscopy surgery were included and randomly divided into group ISB (n=23) and group ST (n=23). Patients in group ISB received 10 ml 0.25% ropivacaine on the lateral side of C5 and C6. Patients in group ST were treated with 5 ml 0.25% ropivacaine on both sides of the superior trunk of brachial plexus. The diaphragmatic excursion, Numerical Rating Scale(NRS), duration of the block, handgrip strength were recorded at different time. Results No statistical difference was detected between the two groups in the reduction of diaphragmatic excursion within 30 min after block (P>0.05). Compared with ISB patients, ST patients had significantly less diaphragmatic excursion at 3 h after block（P<0.05). 30 minutes after block, 8.7% patients in ISB group reached complete HDP and 52.2% patients reached partial HDP. At the same time, no complete HDP and 26.1% partial HDP were detected in ST group. 3 hours after block, patients in ST group had lower complete HDP rate (0.0% vs 17.4%) and lower partial HDP rate (39.1% vs 65.2%) than patients in ISB group. At 30 minutes and 3 h after block, the reduction of grip strength in ST group was significantly lower than that in ISB group (P<0.001). ST group had lower NRS than ISB group (P<0.05). The average block time in ISB group (8.3±1.97 )h was significantly lower than that in ST group (10.9±1.26)h (P<0.01). Conclusion Superior trunk block with 10 ml 0.25% ropivacaine is superior compared to interscalene brachial plexus block in occurrence of HDP, decrease of grip strength, postoperative pain and block duration.

Oxycodone versus sufentanil in patients undergoing laparoscopic gastrointestinal surgery / 药学实践杂志

Objective To compare the perioperative application of sufentanil and oxycodone in patients undergoing laparoscopic surgery for gastric or colorectal cancer. Methods 59 patients were selected and randomly divided into group O and group S. Anesthesia was induced with sufentanil 0.3 μg/kg in group S and oxycodone 0.3 mg/kg in group O. Anesthesia was maintained with sevoflurane balanced anesthesia. When heart rate or blood pressure reached 20% over the baseline, additional dose of oxycodone 0.1 mg/kg was given in group O and sufentanil 0.1 μg/kg in group S. 30 minutes before the end of surgery, patients in group S received sufentanil 0.1 μg/kg and group O with oxycodone 0.1 mg/kg separately. Two hours in the PACU, a rescue dose of sufentanil 0.1 μg/kg or oxycodone 0.1 mg/kg was given to the patients with VAS score bigger than 4. Hemodynamic index, VAS score, Ramsay score, adverse responses and analgesics rescue were recorded. Results No difference was found in hemodynamic index, VAS score and analgesics rescue between the two groups (P>0.05). Ramsay score of group S is lower than that of group O (P=0.014). Induction period bucking incidence in group O was obviously lower than that in group S(P=0.002). The incidence of emergency agitation in group O was significantly lower than that in group S(P=0.045).There was no significant difference in respiratory depression, postoperative nausea and vomiting between two groups (P>0.05). Conclusion Compared with sufentanil, oxycodone significantly reduced the incidence of bucking and emergency agitation. Oxycodone provided better sedation to patients who received laparoscopic surgery for gastric or colorectal cancer.

Novel mutations p.Lys327X and p.Leu424CysfsX8 underlying congenital factor Ⅺ deficiency / 中华医学遗传学杂志

OBJECTIVE@#To analyze the phenotype and genetic mutations in a pedigree affected with factor Ⅺ (FⅪ) deficiency.@*METHODS@#Activated partial thromboplastin time (APTT), FⅪ activity (FⅪ:C) and FⅪ antigen (FⅪ:Ag) were determined for the proband and his family members. All exons and exon-intron boundaries of the FⅪ gene of the proband were analyzed by direct sequencing. Suspected mutation was verified in his family members.@*RESULTS@#The proband had APTT of 82.4 s, FⅪ:C of 0.8%, and FⅪ:Ag of T (Lys327X) mutation in exon 10 and c.1325delT (Leu424CysfsX8) mutation in exon 12 of the FⅪ gene. His elder sister, son, daughter, two granddaughters and one grandson were heterozygous carriers of the c.1033A>T mutation, while his older sister and younger brother were heteozygous carriers of the c.1325delT mutation. Analysis using Mutation Taster software showed that both p.Lys327X and p.Leu424CysfsX8 may affect the function of protein and lead to the corresponding disease.@*CONCLUSION@#The novel mutations of Lys327X and Leu424CysfsX8 of the the FⅪ gene probably underlie the pathogenesis of congenital coagulation factor Ⅺ deficiency in this pedigree.

Rare thalassemia mutations among southern Chinese population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect rare types of thalassemia mutations among southern Chinese population.</p><p><b>METHODS</b>Peripheral blood samples from 327 patients from various regions of southern China were collected. The patients were suspected as rare-type thalassemia for their inconsistency between hematological phenotypes and results of routine mutation screening. The samples were further analyzed with GAP-PCR and DNA sequencing.</p><p><b>RESULTS</b>One hundred and eight cases were diagnosed as rare types of thalassemia. Among whom 10 rare α-globin gene mutations including --THAI, HKα, αααanti3.7, αααanti4.2, -α2.8, -α27.6, CD74 GAC>CAC (Hb Q-Thailand), CD30 (-GAG), CD31 AGG>AAG and CD118 (+TCA), and 12 rare β-globin gene mutations including CD37 TGG>TAG, CD39 CAG>TAG/CD39 CAG>TAG, β II-2 (-T), -90(C>T), -31(A>C), -88(C>T), CD7(-A), CD138(+T), CD89-93 (--AGTGAGCTGCACTG), CD54-58 (-TATGGGCAACCCT), Chinese G γ +(A γδβ)0 and Vietnamese HPFH (HPFH-6) were identified. -88(C>T) (HBB: c.-138C>T) and CD39 CAG>TAG (HBB: c.118C>T) were discovered for the first time in Chinese population. CD7(-A) (HBB: c.23delA) and CD138(+T) (HBB: c.416_417insT) were new types of β-globin gene mutations.</p><p><b>CONCLUSION</b>The present study have enriched the mutation spectrum of thalassemia in southern China, which has provided necessary information for its diagnosis.</p>

The establishment of a high resolution melting analysis based rapid screening system for UGT1A1 gene mutation / 中华检验医学杂志

Objective To develop a high-resolution melting ( HRM ) assay for rapidly screening Gilbert syndrome ( GS) and Crigler-Najjar syndrome ( CNS) associated with UGT1A1 defects.Method Methodology was developed .Then, we applied the established method to analyze 61 clinical samples from neonatal patients with severe unexplained unconjugated hyperbilirubinemia .Neonates with known risk factors for developing hyperbilirubinemia , such as ABO hemolysis, G6PD deficiency, sepsis, hypoxic ischemic encephalopathy were excluded .Five pairs of PCR primers were designed to detect the five common mutations (G211A, C686A, C1091T, C1352T and T1456G) in Asia population.PCR and HRM Assay conditions were optimized.UGT1A1 genotyping in clinical samples was performed by using the established HRM analysis , and all results were subsequently confirmed by direct DNA sequencing .Results The mutants were readily differentiated by using HRM analysis .In this study, 42 neonates were identified with UGT1A1 mutation, and 4 different known variants were detected .Conclusion HRM analysis in this study was economical, convenient, rapid, effective for screening UGT1A1 gene mutations, which can serve as an reliable method for the clinical diagnosis of GS and CNS and the large-scale molecular epidemiological research of UGT1A1 gene-related diseases.

Molecular epidemiology of G6PD deficiency in Chaozhou area of eastern Guangdong Province / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To determine the incidence and molecular characteristics of G6PD deficiency in Chaozhou region of eastern Guangdong Province.</p><p><b>METHODS</b>G6PD enzyme activity was assayed with an auto-bioanalyzer. Reverse dot blotting (RDB) was used for detecting 6 common G6PD mutations. Samples with no mutation detected by RDB were further sequenced for unknown mutations.</p><p><b>RESULTS</b>The rate of G6PD deficiency was 3.36% (142/4224). 2.33% (47/2013) of males and 4.3% (95/2208) of females were affected. 12 mutations were detected among the 142 patients, which included c.1376G>T, c.1388G>A, c.1024C>T, c.392G>T, c.871G>A, c.95A>G, c.517T>C, c.131C>G, c.1376G>T/c.517T>C, c.871G>A/IVS-1193T>C/c.1311C>T, c.1376G>T/IVS-11, 93T>C/c.1311C>T and c.1376G>T/c.486_34delT (rs3216174).</p><p><b>CONCLUSION</b>The incidence of G6PD deficiency in Chaozhou region was lower than that of the Hakka population of Guangdong Province, and the mutation types were diversely distributed in this region. c.1376G>T, c.1388G>A and c.1024C>T were the most common mutations, which was followed by c.517T>C. In addition, c.131C>G has been first discovered in the Chinese population. c.1376G>T/c.517T>C and c.1376G>T/c.486_34delT(rs3216174) were new types of compound heterozygous mutations in females.</p>

Phenotypes and characteristics of human skin-derived precursors / 中国组织工程研究

BACKGROUND:Human skin-derived precursors can be cultured for a long term in vitro, and differentiated into neurons, glial cel s, smooth muscle cel s, Schwann cel s and cel s with peripheral neurons phenotype. OBJECTIVE:To investigate the culture conditions and multiple differentiation capacity of multipotential stem cel s from human skin, especial y the potentials of differentiating into neurons and osteoblasts. METHODS:Human skin-derived precursor cel s were cultured with trypsin digestion method, and identified with immunocytochemistry. Cel s at passages 3-4 were induced to differentiate into neurons and osteoblasts, and underwent von Kossa staining protocol for calcium, chondrocyte induction, toluidine blue staining, immunohistochemical staining and Sudan black staining. The expression of nestin, vimentin,βIII-tubulin, S100 and col agen II in the human skin-derived precursors was detected. RESULTS AND CONCLUSION:The human skin-derived precursor cel s cultured with trypsin digestion method could proliferate and form suspending spheres, and nestin positive cel s were detected at any time point of the culture. Al the cultured cel s expressed vimentin, and some adherent cel s expressedβIII-tubulin. Human skin-derived precursor cel s were induced with Salvia miltiorrhiza to differentiate into neuron-like cel s, and expressed marker of nerve cel s. Skin-derived precursors could be induced to differentiate into osteoblasts and von Kossa staining displayed black calcified nodules in the culture dish. Skin-derived precursors could also be induced to differentiate into chondrocytes, and toluidine blue staining was strongly positive, and some cel s expressed col agen II, which suggested that, the differentiated cel s contained chondrocytes. Experimental findings indicate that, skin contains multipotential stem cel s that are capable of differentiating into osteoblasts, chondrocytes, Schwann cel s and oligodendrocytes.

Culture and identification of human embryo-derived myoblasts / 中国组织工程研究

BACKGROUND:There are myoblasts in human embryonic skeletal muscle. It remains poorly understand whether myoblasts in vitro can form myotube and what are the corresponding markers for identifying myoblasts and myotubes. OBJECTIVE:To investigate whether in vitro cultured myoblasts from human embryonic skeletal muscle can form myotube and whether they can express neural markers. METHODS:Human embryonic muscle-derived myoblasts were cultured in serum-containing medium. When the primary culture was established, cultured cel s were identified with immunocytochemistry for neural markers, such asβ-tubulin markers (desmin, myogenin, smooth muscle actin and myosin). RESULTS AND CONCLUSION:A population of myoblasts could migrate from human embryonic muscle tissues. They could express the markers for skeletal muscle such as desmin and myogenin, and they could express neuron specific enolase, nestin and neurofilament 200. They could form myotubes in vitro, and myotubes expressedβⅢ-tubulin, neurofilament 200 and glial fibril ary acidic protein. The data support the hypothesis that myoblasts from human embryonic muscle express neural markers and muscle markers, and cultured myoblasts and myotubes expressed neuron specific enolase,β-tubulin Ⅲ, nestin, neurofilament 200 and glial fibrillary acidic protein. This indicates that these markers could not be used for cel identification of trans-differentiation study from muscle origin to nervous system.

In vitro differentiation of human bone marrow stromal cells into neural stem cells in serum-free medium / 中国组织工程研究

BACKGROUND:At present,the most frequently agents used for neural induction of bone marrow stromal cells(BMSCs)in vitro are anti-oxidants,such as beta-mercaptoethanol and all trans-retinoids.The majorities of induction from BMSCs are neuron-like cells in these protocols;however,whether it has neuronal function or not should be further studied.OBJECTIVE:TO investigate the differentiated characteristics of inducing human BMSCs into neural cells in serum-free medium.DESIGN:Observational study.SETTING:Chaozhou Central Hospital.MATERIALS:The experiment was carried out in the Chaozhou Central Hospital from April 2004 to December 2005.Adult bone marrows were derived from femoral and tibial bone marrow of three patients with fracture.All patients provided the confirmed consent and were approved by the Ethics Committee of Chaozhou Central Hospital.DMEMIF12 medium(1:1),fetal bovine serum (FBS), glutamine, N2 supplements and B27 Supplements were from GIBCO/BRL Company;recombinant basic fibroblast growth factor(bFGF)and recombinant epidermal growth factor(EGF)from Sigma Company;monoclonal antibody for vimentin(1:100),monoclonal antibody for myelin basic protein(MBP) (1:100),monoclonal antibody for S1 00(1:1 00),monoclonal antibody for neuron specific enolase(NSE)(1:1 00),and monoclonal antibody for neurofilament 200(NF200)(1:1 00)from Beijing Zhongshan Company;monoclonal antibody for glial fibrillary acidic protein (GFAP)(1:200)and polyclonal antibody for nestin(1:100)from Boster Company(Wuhan);mouse monoclonal antibody for beta-tubulin 3(1:1 000)from Sigma Company;SP-9000 kits and quick AEC from Beijing Zhongshan Company; culture dishes and flasks from Coming Company.METHODS:BMSCs from human bone marrow were cultured in serum-containing medium.When the primary culture was established, BMSCs were transferred into serum-free medium containing N2 or B27 supplement with 20 μg/L basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF),and cells were cultured in an incubator containing C02of 0.05 volume fraction at 37℃. Morphological changes of BMSCs in serum-free medium were observed under phase contrast microscope. And two days after culture. Expression of relative markers of BMSCs was detected withimmunocytochemistry.MAIN OUTCOME MEASURES:Morphological changes of BMSCs and expression of relative markers of nerve cells.RESULTS:A population of BMSCs could be isolated from adult human bone marrow,and they were processed to obtain a fibroblast-like population and were expanded as undifferentiated cells in culture for more than 10 passages.indicating their proliferative capacity.They could form spheroid state when they were sub-cultured in serum-free media supplemented with bFGF and EGF.these cells could express the markers for neural stem cells such as vimentin and nestin;they could expressed neuron specific enolase(NSE),beta-tubulin 3,TrkC and neurofilament 200(NF200)when they were plated on dishes with serum-containing medium; some cells exhibited the phenotypes for astrocytes.expressing gilal fibrillary acidic protein(GFAP)and S100 protein.CONCLUSION:The morphology,protein expression and differentiation ability of BMSCs in serum-free medium was similar to those of neural stem cells.The data support the hypothesis that adult bone marrow contains stem cells capable of differentiating into neural cells,the serum-free media make BMSCs overcome their mesenchymal commitment,showing the phenotypes for neural stem cells.

Long-term culture and differentiation of skin-derived mesenchymal stem cells / 生物医学工程学杂志

The aim of this study was to investigate the culture conditions of skin-derived mesenchymal stem cell (sMSCs) and to explore a new cell source for central nervous system cell transplantation. The cells from skins of mice were primarily isolated and cultured in serum-free medium, and they were transferred into serum-containing medium after passaged 2, and the passaged cells were identified by immunocytochemistry and induced to differentiate into multiple lineages. The results indicated that a population of sMSCs could be isolated from skins, they could be maintained in vitro for extended periods with stable population doubling, and they were expanded as undifferentiated cells in culture for more than 10 passages, indicating their proliferative capacity. About 60% of sMSCs expressed vimentin and the majorities of these cells expressed fibronectin. They could differentiate into adipocytes, osteogenic cells and fibroblast-like cells, they could differentiate into neurons with a simple protocol, and almost 50-60% of these cells expressed neuron specific enolase (NSE) and neurofilament (NF); and the differentiated neurons showed typical complicated morphology of neurons. In conclusion, skin contains stem cells that are capable of multiple differentiation; they could be cultured in vitro for long time and could maintain their characteristics of stem cells, and they may represent an alternative autologous stem cell source for CNS cell transplantation.

Long-term culture and differentiation of neural stem cells of embryonic mice / 中华外科杂志

<p><b>OBJECTIVES</b>To assess the culture and differentiation of neural stem cells in embryonic mice and set up a basis for further research in to neural stem cells.</p><p><b>METHODS</b>Embryonic cortices of mice were dissociated and single cell suspensions were achieved by mechanical methods in sterile conditions, and cells were seeded in uncoated plate in N2 medium. The cells were passaged by mechanical methods, frozen and thawed by general procedure. They were identified by immunocytochemical techniques.</p><p><b>RESULTS</b>Neural stem cells from embryonic mice were successfully cultured forming typical neurospheres in suspension. Neurons, astrocytes and oligodendrocytes were differentiated from neural stem cells, with a ratio of 7%, 85% - 90% and 2% - 4% respectively.</p><p><b>CONCLUSIONS</b>Neural stem cells, which can be cultured and passaged steadily in vitro and they are the ideal cell sources for cell transplantation and gene therapy.</p>

Human adipose tissue-derived stromal cells differentiate into osteogenic cells / 实用口腔医学杂志

Objective: To investigate the optimal culture conditio ns for adipose tissue-derived stromal cells(ADSCs) and for the induction of these cells to differentiate into osteogenic cells. Methods: ADSCs were cultured with routine methods,bFGF at 20 ng/ml was added into the medium and the proliferative of ADSCs was examined by cell counting. 0.1 ?mol /L of dexamethasone,10 mmol/L of ?-glycerophosphate and 50 ?mol/L of ascorbic acid were adapted to induce the cells to differentiate into osteogenic cells, ADSCs were identified by immunocytochemistry and differentiated osteogenic cells were identified by alkaline phosphatase(AP) staining and immunocytochemistry. Result: A population of ADSCs could be isolated from adul t human adipose tissue,the cells were fibroblast-like and could be maintaine d in vitro for extended periods with stable population doubling.The cells w ere expanded as undifferentiated in culture for more than 10 passages, indicati ng their proliferative capacity.bFGF stimulated the cell proliferation.Dexameth asone,?-glycerophosphate and ascorbic acid induced (40?8.6)% of ADSCs to ex press alkaline phosphatase(AP) ,(35?10.6)% of AP positive ADSCs were found to be collagen I positive. Calcification plaques were occasionally found in the cul tures. Conclusion:The data support the hypothesis that adu lt human adipose tissue contains stem cells capable of diffferentiating into ost eogenic cells.

Culture of skin-derived precursors and their differentiation into neurons / 中华器官移植杂志

ObjectiveTo investigate the culture conditions of skin-derived precursors (SKPs) and to explore a new cell source for central nervous system cell transplantation.MethodCells from skins of juvenile and adult mice were isolated and cultured in serum -free medium, and mechanical methods were adapted to passage these cells and ce lls were identified by immunocytochemistry.ResultsA population of SKPs could be isolated from adult and neonatal skins. They co uld be maintained in vitro for extended periods with stable population doubling, and they were expanded as undifferentiated cells in culture for more than 8 pas sages, indicating their proliferative capacity. About 50?% of SKPs expressed n estin and the majorities of these cells expressed fibronectin when they were pla ted on polyornithine and laminin coated plates. About 5?% cells showed typical complicated neuronal states and expressed NF-M and NSE when SKPs were plated i n serum-containing medium. These cell could also differentiate into adipocytes and fibroblast-like cells.ConclusionsAdult skin contains stem cells capable of differentiating into neurons, adipocyt es and fibroblast-like cells. SKPs may represent an alternative autologous stem cell source for CNS cell transplantation.