Clinical application of blood adsorption / 中国小儿急救医学

Blood adsorption, one of the blood purification, can be classified into hemoperfusion and immunoadsorption.In recent years, double plasma molecular adsorption, a combined adsorption, has also been widely used in clinical practice.Based on adsorption, the toxins in blood of patients can be efficiently removed by hemoadsorbents.There are two kinds of adsorbents commonly used in hemoperfusion: carbon and resin, and two types of adsorbents in immunoadsorption: biological affinity and physicochemical affinity.Adsorption has been widely applied in clinical practice, involving in sepsis, organ transplantation, systemic lupus erythematosus, liver failure, autoimmune diseases and so on.The review described the application of blood adsorption in clinical practice.

Evaluation of the construction status of a healthy city in central China based on the health index method / 公共卫生与预防医学

Objective To evaluate the construction status of a healthy city in central China, and clarify the weak links and construction results in the construction of a healthy city, and to provide a basis for the relevant government departments to issue policies. Methods Healthy city construction evaluation system in this city was established according to the National Healthy City Construction Evaluation System (2018) of the National Healthy City Evaluation Office Forty-two sets of health city construction index data in 2017-2018 were collected and analyzed. The single index was calculated according to the Guidance Manual for Filling in the Healthy City Evaluation Data, and the single index data was then standardized. The Delphi method was used to consult the weights of the three-level index. Finally, the sub-index and the total index of each dimension of the city's healthy city construction were calculated based on the health index method. Results The results of the 2017-2018 healthy city construction evaluation showed that 35 of the 42 indexes participated in the evaluation, and the total-indices of the two years were 77.71 and 79.95, respectively. The two-year sub-indices of the five dimensions of health service, health culture, health environment, healthy population, and healthy society were 17.17 and 18.32, 11.39 and 12.85, 19.21 and 17.94, 13.81 and 13.81, 16.12 and 17.04, respectively. Except for the decline in the health environment dimension, the other four sub-indices and the total healthy city index showed an upward trend year by year. In the past two years, the comprehensive proportions of the corresponding weight of the five dimensions sub-indices were 95.58% in health culture, 95.47% in health service, 73.99% in health environment, 73.27% in healthy society, and 69.52% in healthy people. Conclusion After the construction of national healthy city and the pilot construction of national healthy city, a city in central China achieved its first results in five dimensions of healthy city. In 2018, among the 35 indexes participated in the evaluation in this city, 30 were positive, and 26 were better than the national/ provincial target value. However, some construction indices were still far from the standard values. The development of the five dimensions was unbalanced, and there were weak links in different degrees.

Standardized application of minimally invasive pars plana vitrectomy for complicated cataract / 中华实验眼科杂志

Phacoemulsification, an effective surgical method in the treatment of cataracts, is widely used in clinics.Traditional cataract surgery is difficult, risky and has a poor prognosis for special intractable cataract patients.Minimally invasive pars plana vitrectomy (PPV) is the a preeminent assistive technique to treat cataracts and their relative complications.Application and surgical techniques of combined cataract extraction with PPV in the intractable cataract complicated with malignant glaucoma, posterior capsular defect, post-PPV, ectopia lentis, aphakic eyes without sufficient capsular support, and perioperative complications are discussed, and it is necessary for eye doctors to put into effect a standardized and guiding relevant diagnostic and therapeutic measures.

Clinical Observation of Nasal Continuous Positive Airway Pressure Ventilation in the Treatment of Neonatal Respiratory Failure / 现代医院

Objective To explore the clinical efficacy of nasal continuous positive airway pressure ventilation and conventional mechanical ventilation in the treatment of neonatal respiratory failure (NICU).Methods 68 cases of neonatal respiratory failure in our hospital from June, 2014 to June, 2016 in neonatal intensive care unit, according to the random number table method were divided into two groups: The observation group involved 34 patients with nasal continuous positive airway pressure, the control group 34 patients with conventional mechanical ventilation treatment.The changes of arterial blood gas analysis, clinical effect and complication were compared between the two groups.Results The PCO2 of the two groups was significantly decreased (P0.05).However, pH value was significantly increased, and the observation group was smaller than the control group (P<0.05).The total effective rate was 94.12% in the observation group and 67.65% in the control group (P<0.05).The time of ventilation and incidence of comorbidities in the observation group was significantly shorter than that in the control group (P<0.05).Conclusion Nasal continuous positive airway pressure ventilation for treatment of neonatal respiratory failure can more significantly improve the efficacy than conventional mechanical ventilation, shorten the time on the machine and reduce the relevant complications.So it is worth promoting.

Construction of a high efficient pancreatic duodenal homeobox 1/insulin dual-reporter vector and its preliminary application / 中国组织工程研究

BACKGROUND:Islet beta cell replacement therapy is one of the most promising approaches for treating type 1 diabetes mellitus. However, its large scale application is hampered by a shortage of islet beta cells for transplantation. Pluripotent stem cells are one of ideal seed cells for islet beta cell replacement therapy, but pancreatic beta-cell differentiation is time-consuming and labor-intensive. OBJECTIVE:To construct a high efficient pancreatic and duodenal homeobox 1 (Pdx1)/insulin dual-reporter vector and to monitor the key genes expression during pancreatic beta-cell differentiation from pluripotent stem cells. METHODS:In order to construct a high efficient Pdx1/insulin dual-reporter vector, puromycin resistance gene was firstly introduced into pTiger vector, and then the original 410 bp mouse Ins1 promoter of the vector was replaced by 646 bp mouse Ins1 promoter. Finally, the dual-reporter vector was transduced into INS-1 and human induced pluripotent stem cells to testify its function. RESULTS AND CONCLUSION:The high efficient Pdx1/insulin dual-reporter vector was constructed successfully. The vector successfully acquired puromycin resistance gene and high gene expression efficacy of insulin in INS-1 cells. The specific gene expression pattern of Pdx1/insulin was first found in INS-1 cells. To conclude, the real-time monitoring function of Pdx1/insulin expression is preliminarily confirmed during pancreatic beta-cell differentiation.

A prospective randomized controlled trial of laparoscopic repair versus open repair for perforated peptic ulcers / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To compared the clinical efficacy of laparoscopic repair (LR) versus open repair (OR) for perforated peptic ulcers.</p><p><b>METHODS</b>From January 2010 to June 2014, in Shanghai Tongji Hospital, 119 patients who were diagnosed as perforated peptic ulcers and planned to receive operation were prospectively enrolled. Patients were randomly divided into LR (58 patients) and OR(61 patients) group by computer. Intra-operative and postoperative parameters were compared between two groups. This study was registered as a randomized controlled trial by the China Clinical Trials Registry (registration No.ChiCTR-TRC-11001607).</p><p><b>RESULTS</b>There was no significant difference in baseline data between two groups (all P>0.05). No significant differences of operation time, morbidity of postoperative complication, mortality, reoperation probability, decompression time, fluid diet recovery time and hospitalization cost were found between two groups (all P>0.05). As compared to OR group, LR group required less postoperative fentanyl [(0.74±0.33) mg vs. (1.04±0.39) mg, t=-4.519, P=0.000] and had shorter hospital stay [median 7(5 to 9) days vs. 8(7 to 10) days, U=-2.090, P=0.001]. In LR group, 3 patients(5.2%) had leakage in perforation site after surgery. One case received laparotomy on the second day after surgery for diffuse peritonitis. The other two received conservative treatment (total parenteral nutrition and enteral nutrition). There was no recurrence of perforation in OR group. One patient of each group died of multiple organ dysfunction syndrome (MODS) 22 days after surgery.</p><p><b>CONCLUSION</b>LR may be preferable for treating perforated peptic ulcers than OR, however preventive measures during LR should be taken to avoid postopertive leak in perforation site.</p>

Neuron-like differentiation of bone marrow mesenchymal stem cells from childrenin vitro / 中国组织工程研究

BACKGROUND:In the past, the culture and differentiation of bone marrow mesenchymal stem celsin vitrowere mostly reported in the adult or animal rather than in children. OBJECTIVE: To explore the ability of bone marrow mesenchymal stem cels from children differentiating into neural stem cels and nerve cels. METHODS: Bone marrow mesenchymal stem cels from children were isolated and cultured, and passage 12 cels were cultured in the pre-induction medium (DMEM culture medium containing 10% fetal bovine serum and 1 mmol/L β-mercapto ethanol) and induction medium (DMEM containing 2% dimethyl sulfoxide and 150 μmol/L butylated hydroxyanisole). Expression of nestin and β-tublin III was detected using immunocytochemistry method at 30 minutes and 7 days after induction, while RT-PCR was used to detect nestin mRNA expression at 0, 5.5, 6 days after induction. RESULTS AND CONCLUSION: After combined induction, the cels shrank from round shape to tapered, polygonal or oval shape, and cel processes extended gradualy and became filament-like shape. Interconnected cels formed a network at 6 days after combined induction. The expression of nestin antigen was positive at 30 minutes after induction, while the expression of β-tublin was positive at 7 days. RT-PCR findings showed that positive expression of nestin mRNA was detected at 5.5 hours of induction, and then disappeared at 6 days. These findings show that the combined use of dimethyl sulfoxide and butylated hydroxyanisole can induce bone marrow mesenchymal stem cels from children to differentiate into neural stem cels and nerve cels in vitro.

Establishment of a novel biotin-inducible eukaryotic gene regulation system / 生物工程学报

To establish a gene regulation system compatible with biopharmaceutical industry and gene therapy, we constructed a fusion protein of biotin ligase from Bacillus subtilis (BS-BirA) and the trans-activation domain, and used its expression vector as the regulatory vector. Meanwhile, BS-BirA-specific operators were ligated upstream of attenuated CMV promoter to obtain the response vector. In this way, a novel eukaryotic gene regulation system responsive to biotin was established and named BS-Biotin-On system. BS-Biotin-On system was further investigated with the enhancing green fluorescent protein (EGFP) as the reporter gene. The results showed that our system was superior to the current similar regulation system in its higher induction ratio, and that the expression of interest gene could be tuned in a rapid and efficient manner by changing the biotin concentrations in the cultures, Our results show that the established system may provide a new alternative for the exogenous gene modulation.

Functional research advances of the protection of alpha crystallin to the lens transparency / 中华实验眼科杂志

α-Crytallin,composed of two menbers of the small heat shock protein,is the major component of the cytoplasm in lens.α-Crytallin not only possesses chaperone-like activity,but also plays important roles in regulation of cell cycle,enhancing genome stability and prevention of the stress-induced apoptosis.The gene mutation associated with α-crytallin is a common cause of hereditary cataract,which is the major cause of childhood blindness.Whereas various changes of the α-crytallin can result in the most common cause of blindness in the world—age-related cataract.Understanding of the function of α-crytallin helps to comprehend the development of lens and how the lens maintain its normal function,and to provide a theoretical basis for the prevention and treatment of the cataract related to the malfunction of α-crytallin.The genetic mapping and mechanism of cataract and the function of α-crytallin are summarized here.

Evaluation of the critical process parameters for the cultivation of recombinant Chinese hamster ovary cells in serum-free fed-batch mode / 生物工程学报

Taking a suspension adapted recombinant CHO cell line, 11G-S expressing human Pro-urokinase (Pro-UK) as the object of study, the impacts of different feeding nutrients, the start time of feeding and cell inoculation density on the growth and Pro-UK production of 11G-S cells in serum-free fed-batch culture were evaluated in 100 mL shacking flasks. The results indicated that amino acids, serum-free supplements and inorganic salts played important role in cell growth, cell viability and protein expression. And the effects of cells fed-batch culture was much better with the initial cell inoculation density at 3 x 10(5)-4 x 10(5) cells/mL and the start time of feeding set at 72 h, a maximum viable cells density of 7.8 x 10(6) cells/mL with a peak Pro-UK activity at 8570 IU/mL was achieved during 12 d fed-batch culture. Further, the mu of the 11G-S cells at the middle phase of the fed-batch culture, and both the q(glu) and q(gln) of the 11G-S cells at the middle and later phases of the fed-batch culture was higher than that of the 11G-S cells at the same phase of the batch culture, respectively.

Analysis of the transcriptional profiling of cell cycle regulatory networks of recombinant Chinese hamster ovary cells in batch and fed-batch cultures / 生物工程学报

In the light of Chinese hamster ovary (CHO) cell line 11G-S expressing human recombinant pro-urokinase, the differences of gene expression levels of the cells in different growth phases in both batch and fed-batch cultures were revealed by using gene chip technology. Then, based on the known cell cycle regulatory networks, the transcriptional profiling of the cell cycle regulatory networks of the cells in batch and fed-batch cultures was analyzed by using Genmapp software. Among the approximate 19 191 target genes in gene chip, the number of down-regulated genes was more than those of up-regulated genes of the cells in both batch and fed-batch cultures. The number of down-regulated genes of the cells in the recession phase in fed-batch culture was much more than that of the cells in batch culture. Comparative transcriptional analysis of the key cell cycle regulatory genes of the cells in both culture modes indicated that the cell proliferation and cell viability of the cells in both batch and fed-batch cultures were mainly regulated through down-regulating Cdk6, Cdk2, Cdc2a, Ccne1, Ccne2 genes of CDKs, Cyclin and CKI family and up-regulating Smad4 gene.

Evaluation and application of exogenous gene expression system based on retroviral vector / 生物工程学报

Currently, exogenous gene expression system based on retroviral vector has been widely used as efficient gene expression system in both gene therapeutic research and RNA interference. In this study, we evaluated the efficiency of exogenous gene expression mediated by the retroviral vector in mammalian cells. First, we constructed EGFP (enhanced green fluorescent protein) vector using pcDNA3.1(+) and retroviral vector pQCXIN as backbone vector respectively. Then, we transfected or infected HEK293 cells and CHO-K1 cells with above vector or corresponding retroviral virus, and measured the relative fluorescence intensity (RFI) of EGFP. The results showed that the RFI of the retroviral virus-infected cells was two times higher than that of the plasmid-transfected cells. Further experiments revealed repeated virus infection enhanced the expression of EGFP markedly, with RFI increasing twice after four rounds of virus infection. Furthermore, the EGFP expression in HEK293 cells mediated by the retroviral vector was more stable than transfected with plasmid pcDNA3.1(+). Finally, we further validated the efficiency of exogenous gene expression system based on the retroviral vector by expressing recombinant human activated protein C (rhAPC) in HEK293 cells. We obtained HEK293 cell lines with rhAPC expression between 10 and 15 microg/(10(6) cells d). In conclusion, the exogenous gene expression system based on the retroviral vector is an alternative method for the generation of stable and high-expressing mammalian cell lines.

Construction and characterization of cardiac specific promoter-driven expression vector / 生物工程学报

We constructed and identified cardiac-specific a-myosin heavy chain (alpha-MHC) promoter-driven expression vector. alpha-MHC promoter was amplified by PCR by using mouse genomic DNA as template and inserted into pGEM-T Easy vector. The inserted fragment was released by enzyme digestion, and then the cytomegalovirus (CMV) promoter in pcDNA3.1(+)-EGFP-hygro vector was replaced by the alpha-MHC promoter to construct alpha-MHC-EGFP expression vector. After identification with enzyme digestion, alpha-MHC-EGFP was transfected into mouse primary cardiomyocytes by electroporation. Green fluorescence could be observed in transfected cardiomyocytes, but not in transfected non-cardiomyocytes. Alpha-MHC-EGFP expression vector was specifically expressed in cardiomyocytes, and could be used to purify embryonic stem cell-derived cardiomyocytes.

Analysis of nine cases of acute thallium poisoning / 华中科技大学学报（医学）（英德文版）

In this study nine cases of thallium poisoning in a series of homicidal poisoning were analyzed in order to provide more information concerning thallium poisoning. It was found that the most common clinical feature of thallium poisoning was peripheral neuropathy and paraesthesia was more common than amyasthenia. Understanding of these clinical characteristics of thallium poisoning was helpful to early identification and differential diagnosis. Since the early administration of Prussian Blue, as a specific antidote for thallium poisoning, can substantially improve the prognosis, it is of great importance to establish a correct and early diagnosis.

Analysis of Nine Cases of Acute Thallium Poisoning / 华中科技大学学报（医学）（英德文版）

In this study nine cases of thallium poisoning in a series of homicidal poisoning were analyzed in order to provide more information concerning thallium poisoning. It was found that the most common clinical feature of thallium poisoning was peripheral neuropathy and paraesthesia was more common than amyasthenia. Understanding of these clinical characteristics of thallium poisoning was helpful to early identification and differential diagnosis. Since the early administration of Prussian Blue, as a specific antidote for thallium poisoning, can substantially improve the prognosis, it is of great importance to establish a correct and early diagnosis.

In vitro differentiation of bone marrow mesenchymal stem cells induced by activin A and all-trans retinoic acid into insulin-producing cells / 中国组织工程研究

BACKGROUND: The increasing prevalence of diabetes mellitus has been become one of the diseases which threaten the heath of human being in the 21st century. Islet transplantation is considered to be the most effective approach to cure type Ⅰ diabetes mellitus. However, lack of donor tissue limits the application of this therapy. However, recent progress of stem cell research shows that stem cell therapy may be a potential means to solve this problem.OBJECTIVE: To take activin A and all-trans retinoic acid (AR) in inducing the differentiation of bone marrow mesenchymal stem cells (MSCs) and explore its possibility DESIGN: A randomized controlled experiment.SETTING: Institute of Biotechnology, Academy of Military Medical SciencesMATERIALS: This experiment was conducted at the Institute of Biotechnology, Academy of Military Medical Sciences from November 2004to June 2005. Six male Sprague-Dawley rats, with body mass of 150-160g, were provided by the Experimental Animal Center of Academy of Military Medical Sciences.METHODS: Femoral bone marrow of the rats was extracted under aseptic condition. Bone marrow mesenchymal stem cells (MSCs) were isolated with density gradient centrifugation. Passaged MSCs were randomly divided into 4 groups: high concentration of glucose (HG), AR, beta-mercaptoethanol (ME) and negative control groups. MSCs were induced to differentiate into IPCs with conditional medium containing high concentration glucose, activin A, RA and ME etc. After induction, phenotypes of differentiated cells were examined by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: Expression of insulin and glucagon of differentiated cells were examined by immunocytochemistry. Insulin-1 mR-NA expression of differentiated cells was detected by RT-PCR.RESULTS: After bone marrow mesenchymal stem cells were induced,there were scattered insulin-and glucagon-positive cells in the HG group,many insulin-and glucagon-positive cells in the AR and ME groups, and these cells formed insulin-like structure. The expression of insulin-1mRNA could be observed in the HG, AR and ME groups. Insulin-and glucagonpositive cells and the expression of insulin-1mRNA were not observed in the negative control group.CONCLUSION: We adopt an induction scheme based on AR and other matured factors, and successfully make bone marrow mesench.ymal stem cells induce and differentiate into insulin positive reaction cells and form insulin-like structure, but its induction efficiency needs further improvement.

The committed differentiation of bone marrow mesenchymal stem cells into tenocytes / 解放军医学杂志

Objective To investigate the feasibility of committed differentiation of rhesus bone marrow mesenchymal stem cells (MSCs) into tenocytes induced by BMP12. Methods MSCs were transfected with pTARGETTM bearing BMP12 gene by electroporation. The transfected cells were identified by morphological observation and molecular biological measure. Results Under the observation of light microscope, the morphological features of transfected cells changed significantly compared to parental MSCs. RT-PCR data showed the transfected cells had the mRNA expression of BMP12 and collagen Ⅰ, but without that of collagen Ⅲ. 98.39% of the transfected cells were CD44+ and negative for HLA-DR. Conclusion BMP12 could induce MSCs into tenocytes, and bone marrow MSCs might be the optional seed cells for tendon tissue engineering.

Cloning of integral mature peptide gene of human GDF-5 / 华中科技大学学报（医学）（英德文版）

The integral mature peptide gene of human growth differentiation factor-5 (GDF-5) was cloned to provide the essential foundation for study on the biological characteristics of GDF-5 at gene and protein levels. Two primers were chemosynthesized according to the hGDF-5 sequence reported in Genbank. The hGDF-5 gene was gained by RT-PCR methods from the total RNA extracted from human fetus cartilage tissue, and was cloned into vector pMD18-T. The sequence of recombinant plasmid pMD18-T-hGDF-5 was analyzed by sequence analysis. DNA agarose gel electrophoresis showed that the product of RT-PCR was about 380bp, and double enzyme digestion of the recombinant plasmid corresponded with it. The result of sequence assay was in agreement with the reported hGDF-5 sequence in Genbank. Our results showed that the integral mature peptide gene of human GDF-5 was cloned successfully from human fetal cartilage tissue, and totally identified with the sequence of human GDF-5 in Genbank.

Transient expression of BMP12 gene transfer to mesenchymal stem cells / 解放军医学杂志

Objective To investigate the feasibility of the committed differentiation of mesenchymal stem cells(MSCs) into tenocytes,the present study was carried out. Method NMP12 gene transfer to MSCs was performed with pEGFP C1 expressing vector by electroporation. Results Green fluorescence protein(GFP) expression was detected as early as 6h postelectroporation, peaked at 12h and could maintain 3 days. And then, GFP expression of some cells was weakened. Conclusion The results of the present study indicated that the successful transfection of MSCs was made, an the committed differentiation of MSCs into tenocytes was practicable

THE BIOLOGICAL FEATURES OF RHESUS BONE MARROW MESENCHYMAL STEM CELLS / 解剖学报

Objective In order to explore the biological features of bone marrow mesenchymal stem cells. Methods The ALPase activity, Vimentin and Laminin were detected by immunocytochemistry. Cell cycle and surface antigenic features were analyzed by flow cytometry technique. Cell growth curve was measured by MTT test. The telomerase activity was detected by in situ hybridization. Results The ALPase activity and Laminin were negative, Vimentin was positive in the MSCs. The percentage of G1, S, and G2 of cell cycle was 89.5%,7.8%,7.7%, respectively. MSCs were positive for CD166、CD29、CD44、CD105, and negative for CD34 and HLA-DR. The average doubling time of cells was 32 hours. The telomerase activity was positive. Conclusion The cultured cells were not hemopoietic stem cells or fibroblasts, but the MSCs that had not differentiated. Furthermore, it had relatively high proliferation ability.