Objective To construct the recombinant adenoviros containing heat shock protein70 (Hsp70) gene driven by carcinoembryonic antigen (CEA) promoter. Methods Hsp70 gene and CEA promoter were amplified by RT-PCR and PCR, and then subcloned into the shuttle vector pDC316 to construct the recombinant vector PDC316-pCEA-Hsp70. The recombinant vector was co-transfected with adenoviral backbone plasmid into HEK293 cells to generate the recombinant adenovirus Ad5-pCEA-Hsp70. The recombinant adenovirus was purified by CsCl banding and titrated by 50％ tissue culture infective dose (TCID50) assay. After transfection of the recombinant adenovirus into human pancreatic cell lines SW1990 and BxPC3, the expression of mRNA and protein level of Hsp70 were determined by RT-PCR and ELISA,respectively. Results Digestion and DNA sequencing certified that the Hsp70 gene and CEA promoter was successfully inserted into pDC316 plasmid. Virus acquired through co-transfection with backbone plasmid was confirmed to be constructed successfully by PCR amplification. The particles finally expressed was 2.2 ×1011vp/ml, and the titer was 1.5 x 1010 PFU/ml. BxPC3 cancer cells with positive CEA expression showed increased expression of Hsp70 mRNA and protein after infected by recombinant adenovirus; while SW1990 cancer cells with negative CEA expression showed no change of expression of Hsp70 mRNA and protein after infected by recombinant adenovirus. Conclusions The recombinant adenovirus Ad5-pCEA-Hsp70 which can express Hsp70 gene in CEA positive cancer cells is constructed successfully.

Objective To apply quality control circle (QCC) to reducing non-timely maintenance rate of medical equipment.Methods QCC was established,and some measures were taken to observe its effect in decreasing non-timely maintenance rate of medical equipment.Results The non-timely maintenance rate fell from 12.1％ to 4.5％,and 0.3 million Yuan was saved for the manpower cost.Conclusion QCC decreases the non-timely maintenance rate of medical equipment,increases the efficiency of the maintainer,shortens the downtime of the fault medical equipment,enhances hospital economic benefit and patient satisfaction and provides references for other hospitals.

Objective To extract the texture features of endoscopic ultrasonography (EUS) by digital imaging processing(DIP) and pattern recognition,and then to investigate its value for differential diagnosis between pancreatic cancer and chronic pancreatitis.Methods Two hundred and two patients with pathologicaly confirmed pancreatic malignancy,who underwent EUS from Feb 2005 to Mar 2011,and 104 patients with chronic pancreatitis (including 34 cases of autoimmune pancreatitis),who underwent EUS from May 2002 to Aug 2011,were randomly recruited in this study.The optimal texture features of EUS images in this study were selected by the sequence forward search (SFS) algorithm.With the optimal feature combination,cases were automatically divided into pancreatic cancer and chronic pancreatitis based on the findings of support vector machine (SVM),which were compared with the real results.the sensitivity,specificity,accuracy,positive predictive value and negative predictive value were calculated.Results Nine categories and 105 texture features were extracted based on all EUS images,and 13 features were chosen as optimal combination.Images of 306 cases were randomly divided into training set ( 153 cases,101 cases of cancer,52 cases of chronic pancreatitis) and testing set ( 153 cases,101 cases of cancer,52 cases of chronic pancreatitis).The classifier was trained with the training set and tested with testing set.We proceeded 200 times randomly.the average accuracy,sensitivity,specificity,positive predictive value and negative predictive value were ( 86.08 ± 0.14) ％,(79.47 ± 0.32) ％,( 89.71 ± 0.18 ) ％,( 81.21 ± 0.26 ) ％,( 88.93 ± 0.14 ) ％,respectively.Conclusion Differential diagnosis of pancreatic cancer from chronic pancreatitis by Computer-assisted EUS image analysis,highly accurate,convenient,non-invasive and less costly,is a novel and valuable method of early diagnosis.

Abstract: Objective　To investigate the molecular characteristics and drug resistance of non-O1/non-O139 Vibrio cholerae in Zhongshan City, and to provide laboratory basis for cholera prevention and control. Methods　The strains of non-O1/non-O139 Vibrio cholerae isolated from sporadic patients and aquatic products from 2015 to 2021 in Zhongshan city were collected. The identification and cluster analysis of the strains were analyzed by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), the ctxA virulence gene of strains were detected by real-time fluorescence quantitative PCR, the cluster analysis of the strains was analyzed by pulsed-field gel electrophoresis (PFGE), and the drug resistance of the strains were analyzed by microbroth dilution method. Results　From 2015 to 2021, 33 strains of non-O1/non-O139 Vibrio cholerae were isolated from Zhongshan City, including 28 strains from sporadic patients and 5 strains from aquatic products. Through MALDI-TOF-MS identification, 33 strains of non-O1/non-O139 Vibrio cholera can be identified to the level of species, and the identification results were all Vibrio cholerae. Among 33 non-O1/non-O139 Vibrio cholerae strains, 1 strain carried the ctxA virulence gene. The drug-resistant strains accounted for 69.7% (23/33), and the multidrug resistant strains accounted for 18.2% (6/33). A total of 7 kinds of drug resistance spectrum were produced, including 3 kinds of multidrug resistant spectrum, and showed drug resistance to 8 antibiotics, among which the resistance rates to streptomycin, cefazolin and compound sulfamethoxazole were above 30%. The 33 strains of non-O1/non-O139 Vibrio cholerae were divided into 32 PFGE fingerprints with a similarity ranging from 61.7% to 100%. MALDI-TOF-MS cluster analysis divided 33 non-O1/non-O139 Vibrio cholerae strains into two clusters. Conclusions The results of molecular typing of non-O1/non-O139 Vibrio cholerae in Zhongshan City presented diversity, and no significant correlation was found between PFGE and MALDI-TOF-MS cluster analysis. The strains demonstrated various degrees of resistance to certain antibiotics, and there were multidrug-resistant and toxigenic strains. Therefore, it is necessary to alert to the harmfulness of non-O1/non-O139 Vibrio cholerae and enhance monitoring.

OBJECTIVETo investigate the clinical features of Mycoplasma pneumoniae pneumonia (MPP) among children of different ages.

METHODSRetrospective analysis was performed on the clinical data of 112 children who were hospitalized due to MMP between January 2010 and December 2011. The children were divided into 3 groups according to their ages: infants (<3 years; n=20), preschool-aged children (≥3 years; n=41), and school-aged children (6-15.2 years; n=51). The three groups were compared in terms of their clinical symptoms, pulmonary signs, chest X-ray findings and laboratory test results.

RESULTSThe infant group presented mainly with expectoration and wheezing, accompanied by low fever. They showed gastrointestinal symptoms as the most common extra-pulmonary manifestation and had evident pulmonary signs. The majority of the school-aged children group presented with high fever and a severe dry cough, and wheezing was seen in several of them. They showed rash as the most common extra-pulmonary manifestation and had slight pulmonary signs. The symptoms of the preschool-aged children group were in between. In the infant and preschool-aged children groups, most showed bronchopneumonia on chest X-ray, while in the school-aged children group, chest X-rays mostly showed segmental parenchymatous infiltration. The infant group had a higher lymphocyte count than the school-aged children group, while the school-aged children group had a higher serum C-reactive protein level than the infant group.

CONCLUSIONSThe clinical features of MPP are different among children of different ages, especially between infants and school-aged children.

Adolescent ; Age Factors ; Antibodies, Bacterial ; blood ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Pneumonia, Mycoplasma ; diagnosis ; diagnostic imaging ; drug therapy ; Radiography, Thoracic ; Retrospective Studies

The study was aimed to establish a protocol of isolating and culturing adult mesenchymal stem cells (MSC) from human bone marrow aspirate and identify them by surface antigen analysis and committed differentiation in order to provide an experimental foundation for achieving a therapeutic benefit in applying MSC in hematopoietic stem cell transplantation. MSCs were obtained from fresh human bone marrow aspirate by gradient centrifugation with Percoll (1.073 g/ml) and anchoring culture in L-DMEM with 10% fetal bovine serum by a full medium exchange every 3 days. The MSC surface antigens, including CD34, CD45, CD73, CD105, CD166, were analyzed on FACScan flow cytometer. Under culture in conditioned medium for osteogenesis (the hormone cocktail containing 0.1 micromol/L dexamethasone, 10 mmol/L glycerol-2-phosphate and 50 micromol/L ascorbic acid) and adipogenesis (the cocktail containing 1 micromol/L dexamethasone, 5 mg/L insulin, 0.5 mmol/L 1-methyl-3-isobutylxanthine and 60 micromol/L indomethacin), MSCs committedly differentiated into osteoblasts and adipocytes. The differentiated mesenchymal stem cells were identified by morphological observation and immunohistochemical staining. The results showed that by gradient centrifugation and adhesion culture, MSCs could be isolated and culture-expanded from human bone marrow aspirate. These cells were uniformly negative for CD34, CD45 and positive for CD73, CD105 and CD166. The osteogenic differentiated cells were positive for alkaline phosphatase (ALP) and the adipogenic differentiated cells displayed accumulation of lipid vacuoles, as detected by oil red O. It is concluded that MSC can be isolated and expand-cultured from adult human bone marrow aspirate and committedly differentiate into osteoblasts and adipocytes. MSC primary identification can be accomplished by flow cytometry and induced differentiation. The set of methods in current experiment shows somewhat practical value for basic research and clinical application.
5'-Nucleotidase ; metabolism ; Antigens, CD ; metabolism ; Bone Marrow Cells ; cytology ; Cell Adhesion Molecules, Neuronal ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Separation ; methods ; Endoglin ; Fetal Proteins ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology ; Receptors, Cell Surface ; metabolism

OBJECTIVETo develop a new drug of treating the hepatitis C virus by studing the mechanisms of songzhi pills.

METHODThe four-wee old mice and two-month old rats were chosen to induce interferon.

RESULTThe contents of interferon among the groups treated with songzhi pills were significantly higher than those of the normal group and the model group (P < 0.01),

CONCLUSIONSSongzhi pills may have the function of inducing interferon.

Animals ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Gardenia ; chemistry ; Interferon-gamma ; biosynthesis ; Male ; Mice ; Plants, Medicinal ; chemistry ; Polyporales ; chemistry ; Rats ; Rats, Sprague-Dawley

The objective of this research was to explore whether the number of CD34(+)CD38(+) cells infused affects hematopoietic reconstitution after cord blood transplantation. The number of CD34(+)CD38(+) cells in cord blood was analysed with flow cytometry after freezethawing. The body weight and time for neutrophil and platelet recovery were measured in 20 children with acute leukemia. The results showed that the median number of CD34(+)CD38(+) cells infused was 29.47 (9.85 - 325.71) x 10(4)/kg. A median time for neutrophil recovery (> 5 x 10(8)/L) in 20 patients was 18.5 (11 - 32) days, and time for platlet recovery (> 2 x 10(10)/L) in 19 of 20 patients was 45 (12 - 118) days. The number of CD34(+)CD38(+) cells infused correlated with time to neutrophil and platelet recovery (r = -0.577, P < 0.01 and r = 0.503, P < 0.05, respectively). In conclusion, the number of CD34(+)CD38(+) cells infused is correlated with the time for hematologic recovery.
ADP-ribosyl Cyclase ; analysis ; ADP-ribosyl Cyclase 1 ; Adolescent ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Child ; Child, Preschool ; Fetal Blood ; cytology ; transplantation ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia, Myeloid, Acute ; blood ; therapy ; Membrane Glycoproteins ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; therapy

OBJECTIVESeveral studies have shown that L-selectin on CD34-positive cells play a role in hematopoietic reconstitution after peripheral blood stem cell transplantation and allograft bone marrow transplantation. This study sought to investigate whether the numbers of CD(34)(+)CD(62L)(+) cells infused affect the engraftment of hematopoietic stem cells (HSC) and the time to neutrophil and platelet recovery after unrelated umbilical cord blood transplantation for the treatment of childhood acute leukemia.

METHODSTwenty-three children with acute leukemia who received unrelated umbilical cord blood transplantation of mostly mismatched HLA locus were included in this study. Flow cytometry was used to count the numbers of CD(34)(+)CD(62L)(+) cells after freezing-thawing by labelling the cells with anti-CD(34) and anti-CD62L. The patients' clinical data including body weight, engraftment of the HSC, times to neutrophil and platelet recovery were evaluated.

RESULTSTwenty-one patients who received CD(34)(+)CD(62L)(+) cell infusion at a number ranging from 1.37 x 10(5)/kg to 2.68 x 10(6)/kg (median, 3.567 x 10(5)/kg) had successful engraftment of the unrelated umbilical HSC. The numbers of CD(34)(+)CD(62L)(+) cells infused were statistically different between patients who had successful engraftment of the umbilical HSC and those who had not (P < 0.05). The engraftment occurred more commonly in patients who received > 1.3 x 10(5) CD(34)(+)CD(62L)(+) cells/kg. The time of neutrophil recovery (> 500/ microl) ranged from 11 days to 32 days (median, 17.5 days). The data of the time to platelet recovery (> 2 x 10(5)/ microl) were obtained in 18 patients, and it ranged from 12 days to 118 days (median, 14 days). There seemed to be a tendency of correlation between the numbers of CD(34)(+)CD(62L)(+) cells infused and time to platelet recovery (gamma = -0.324, 0.05 < P < 0.1), whereas the numbers of CD(34)(+)CD(62L)(+) cells infused correlated with the time to platelet recovery (gamma = -0.470, P < 0.05).

CONCLUSIONThis study suggests that the numbers of CD(34)(+)CD(62L)(+) cells infused might be involved in the engraftment of HSC and hematologic reconstitution after umbilical cord blood transplantation.

Acute Disease ; Adolescent ; Antigens, CD34 ; blood ; Blood Platelets ; metabolism ; Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Humans ; Infant ; Infusions, Intravenous ; L-Selectin ; blood ; Leukemia ; immunology ; therapy ; Male ; Neutrophils ; metabolism ; Treatment Outcome

Umbilical cord blood (UCB) is an alternative source of hematopoietic stem cells for transplantation with success being associated with the total nucleated cell (TNC) count, CD34(+) cells and colony-forming unit-granulocyte-macrophage (CFU-GM) content infused. This study was purposed to clarify the impact of maternal and neonatal factors on hematopoietic potential of UCB product. UCB samples were screened, processed, tested and cryopreserved according to the Standard Operation Procedure (SOP) of Guangzhou cord blood bank (GZCBB). Relationship of hematopoietic cell parameters with maternal and neonatal characteristics for 4615 UCB units was analyzed retrospectively. The results showed that both collected volume (Mean ± SD: 95.23 ± 22.42 ml; Median: 91.85 ml) and initial TNC [Mean ± SD: (1.34 ± 0.49) × 10(9); Median: 1.25 × 10(9)] correlated well with postprocessed TNC [Mean ± SD: (1.21 ± 0.42) × 10(9); Median: 1.14 × 10(9); p < 0.001], CD34(+)count [Mean ± SD: (5.14 ± 4.55) × 10(6); Median: 4.08 × 10(6); p < 0.001] and CFU-GM content [Mean ± SD: (9.72 ± 8.66) × 10(5); Median: 7.53 × 10(5); p < 0.001]. As for donor factors, only infant birth weight correlated strongly with volume collected and all hematopoietic cell parameters (p < 0.001). UCB samples from bigger babies had higher collected volume, TNC, CD34(+) count and CFU-GM content (p < 0.001). Mother's age had no correlation with all the above parameters. Gestational age correlated positively with initial/postprocessed TNC (p < 0.001) and negatively with CD34(+) count (p = 0.04), but no relation with collected volume and CFU-GM content. Cesarean section produced superior volume (Mean ± SD: 97.05 ± 22.23 ml vs 92.53 ± 22.43 ml; Median: 94.08 ml vs 88.82 ml; p < 0.001), but inferior cell count than vaginal delivery (p < 0.001). Male infants had more initial volume and CD34(+) count (Mean ± SD: 96.41 ± 22.31 ml vs 93.95 ± 22.47 ml; Median: 93.27 ml vs 90.14 ml; p < 0.001); [Mean ± SD: (5.28 ± 5.04) × 10(6) vs (5.00 ± 3.94) × 10(6); Median: 4.18 × 10(6) vs 3.94 × 10(6); p < = 0.042], but lower initial and postprocessed TNC than female ones [Mean ± SD: (1.31 ± 0.50) × 10(9) vs (1.37 ± 0.47) × 10(9); Median: 1.22 × 10(9) vs 1.28 × 10(9); p < 0.001]; [Mean ± SD: (1.18 ± 0.42) × 10(9) vs (1.24 ± 0.41) × 10(9); Median: 1.10 × 10(9) vs 1.17 × 10(9); p < 0.001], while no significant difference of CFU-GM were found between male and female infants. It is concluded that these data may be helpful to optimize the UCB donor selection and improve cost efficiency of UCB bank resource. The heavier infants after vaginal delivery should be selected and large-volume units with higher TNC should be chosen at first.
Adult ; Birth Weight ; Blood Banks ; methods ; Cord Blood Stem Cell Transplantation ; methods ; Delivery, Obstetric ; Donor Selection ; Female ; Fetal Blood ; cytology ; immunology ; Gestational Age ; Hematopoietic Stem Cells ; Humans ; Infant, Newborn ; Male ; Maternal Age ; Pregnancy ; Young Adult