OBJECTIVETherapy effects and cardiac electrical activity comparison of bone marrow stem cells (BMSCs) transplantation and sarcoplasmic reticulum Ca(2+)-ATPase (SERCA2a) gene modified BMSCs transplantation after acute myocardial infarction (AMI) in rats.

METHODSRats with AMI were divided into 4 groups (n = 30) randomly: normal group (n = 6), saline group (control group, n = 8), BMSCs transplantation group (BMSCs group, n = 8) and SERCA2a gene modified BMSCs transplantation group (BMSCs + rAd.SERCA2a group, n = 8). After 14 days, cardiac function was evaluated by echocardiography and heart electrical activity was evaluated by electrocardiogram and microelectrode array (MEA) technology.

RESULTS(1) The transduction ratio of rAd.SERCA2a to BMSCs were 80% to 90%. (2) Left ventricular ejection fraction on 14 days after therapy was significantly higher in BMSCs group and BMSCs + rAd.SERCA2a group than in control group (all P < 0.05). (3) QT duration was significantly shorter [(80.30 ± 6.53) ms vs. (105.31 ± 21.89) ms, P < 0.05] and ventricular premature beats less frequent in BMSCs + rAd. SERCA2a group than in the control group. (4) MEA results suggested that isolated heart beat was significantly slowed down and frequent ventricular arrhythmias and atrioventricular block were recorded in control group. The maximum field potential and field potential duration on infarcted myocardium area in BMSCs group and BMSCs + rAd.SERCA2a group were significantly longer than those in control group[the maximum field potential: (0.51 ± 0.15), (0.55 ± 0.16), (0.23 ± 0.10) mV; field potential duration: (104.5 ± 25.43), (107.67 ± 24.01), (63.00 ± 20.34) ms; all P < 0.05]. (5) The conduction time was the shortest and the cardiac electrical conduction consistency in myocardial infarction tissue was significantly improved in BMSCs + rAd.SERCA2a group.

CONCLUSIONSBMSCs and SERCA2a gene modified BMSCs transplantation could significantly improve cardiac function and BMSCs + rAd.SERCA2a could also effectively improve electrical conduction of infarcted myocardium and attenuate the incidence of arrhythmia after myocardial infarction in rats.

Amino Acid Motifs ; Animals ; Bone Marrow Transplantation ; methods ; Electrocardiography ; methods ; Male ; Microelectrodes ; Myocardial Infarction ; physiopathology ; surgery ; Rats ; Rats, Sprague-Dawley ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; genetics

OBJECTIVETo evaluate the mRNA and protein expressions of peroxiredoxin II (PrxII) in hepatocellular carcinoma (HCC) and their significance.

METHODSHCC was induced by aflatoxin B1 (AFB1) in 6 tree shrews (Tupaia belangeri chinensis). The expression levels of PrxII mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot on HCC tissues and on their surrounding liver tissues (para-HCC). Biopsied liver tissues were taken before the HCC induction (pre-HCC) from the same animals and from a group of blank controlled animals that served as controls. Liver biopsy specimens from 18 cases of human HCC and from 17 healthy human volunteers were studied using the same methods.

RESULTSThe mRNA and protein expressions of PrxII in tree shrew HCC tissues were significantly higher than those in para-HCC and pre-HCC tissues, and also higher than those in the liver tissues from the control animals (all P < 0.05). The expression levels of PrxII mRNA and protein in human HCC tissues were also significantly higher than those in their para-HCC tissues and in the human normal liver tissues (P < 0.05).

CONCLUSIONPrxII might play an important role in hepatocarcinogenesis and might be used as a molecular target for HCC prevention and treatment.

Adult ; Aged ; Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Male ; Middle Aged ; Peroxiredoxins ; genetics ; Tupaiidae

The characteristic pathological changes of Parkinson s disease (PD) include a severe loss of dopamine neurons in the substantia nigra and a severe decrease in dopamine in the striatum. Since the expression of tyrosine hydroxylase (TH) and aromatic L-amino acid decarboxylase (AADC) in the biosynthetic pathway for dopamine are low, a promising approach to the gene therapy of PD is to augment the gene expression of the enzymes in the biosynthetic pathway for dopamine. In the present study, human TH and AADC genes were reconstructed into retrovirous vectors pLHCX and pLNCX(2) respectively. Then pLHCX/TH and pLNCX(2)/AADC were transfected into packaging cell line PA317 with liposome. PA317/TH and PA317/AADC were selected by different antibiotics. Gene expression was examined by methods of immunohistochemistry and in situ hybridization. The catalytic activity of two cloned gene enzymes was assessed in vitro by HPLC-EC. Immunocytochemical staining showed that TH and AADC were expressed efficiently in vitro. Both TH and AADC mRNA were transcripted in PA317 cell lines by using in situ hybridazation. HPLC-EC experiments revealed that the transfected cells produced a significantly higher level of dopamine and L-dopa than the untransfected cells. The two genetically modified cells could improve the production of L-dopa and dopamine in response to suitable substrate. The present results suggest that not only recombinant TH and AADC genes are successfully expressed in vitro, but also the enzymes have respective functional activities. These results have set up a way for in vivo gene therapy of PD with TH and AADC genes.
Aromatic-L-Amino-Acid Decarboxylases ; biosynthesis ; genetics ; metabolism ; Cell Line ; Corpus Striatum ; enzymology ; Dopamine ; biosynthesis ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Humans ; Levodopa ; biosynthesis ; Parkinson Disease ; enzymology ; genetics ; RNA, Messenger ; biosynthesis ; Substantia Nigra ; metabolism ; Transfection ; Tyrosine 3-Monooxygenase ; biosynthesis ; genetics ; metabolism

In order to explore the feasibility of inducing the human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) to differentiate into insulin-secreting cells with biological products alone, hUC-MSCs were separated and purified from the whole umbilical cord by the sequent digestion of collagenase II and trypsin followed by two-step centrifugation. hUC-MSCs were induced with IMDM culture medium containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and Ginkgo biloba extract (GBE). Before and after the induction, the morphological changes were observed under inverse microscope; the islet-related genes were detected by RT-PCR; islet-like clusters (ILCs) were identified by dithizone (DTZ) staining; PDX-1 and immunoreactive insulin (IRI) were examined by immunofluorescence method; the quantity and quality of IRI secretion were assayed by chemiluminescence immunoassay and Western blot respectively. The results showed that the purified hUC-MSCs presented long spindle-like shape and parallel or spiral arrangement which are typical morphological features of MSCs. After the induction, hUC-MSCs changed gradually into round or oval shape and gathered together to form ILCs; there were more than one hundred clusters on the growth surface of a flask of T25; ILCs were stained into positive mauve by DTZ and positive for PDX-1 and IRI; Western blot displayed that most of the IRI was proinsulin (PI). Therefore, hUC-MSCs can rapidly differentiate into insulin-secreting cells under the sole induction of EGF, bFGF, GBE and IMDM, but ILCs are not mature enough to produce sufficient true insulin.
Cell Differentiation ; Cells, Cultured ; Epidermal Growth Factor ; pharmacology ; Fibroblast Growth Factor 2 ; pharmacology ; Ginkgo biloba ; chemistry ; Humans ; Insulin-Secreting Cells ; cytology ; Mesenchymal Stromal Cells ; cytology ; Plant Extracts ; pharmacology ; Umbilical Cord ; cytology

OBJECTIVETo investigate the changes of nitric oxide concentration in the distal portion of a random pattern skin flap and the influence of the exogenous L-arginine on the survival of the random pattern skin flap.

METHODSA random pattern skin flap (7 cm x 2 cm) was cranially designed and elevated on the back of a Wistar rat. An image analysis technology was used to evaluate the survival rate of the skin flap, while a biochemistry method was used to test the concentrations of the NO in the tissue.

RESULTSThe survival area of the flap in the L-arginine-treated group was significantly enlarged (63.83 +/- 5.13)% (P < 0.01) in seven days postoperatively, compared with the control group (43.26 +/- 2.86)%. The NO concentration in the tissue was no statistic difference between all of the groups immediately after the operation (P > 0.05). But, the NO concentration in the control was decreasing at the beginning and then increasing slightly to reach the high level in 12 hours after the operation. It was thereafter slumped down to the baseline in 72 hours after the surgery. Although the changes in the L-arginine-treated group were quite similar to the control excepting of the extent, the NO concentration was kept in a higher level in the sequential time after the operation (P < 0.01, P < 0.01, P < 0.05 and P < 0.01).

CONCLUSIONSThe NO concentration in skin flap tissue after the elevation was going up slightly for a short time. The exogenous L-arginine could promote the NO concentration in the random pattern skin flap to protect it from ischemic injury.

Animals ; Female ; Graft Survival ; drug effects ; Male ; Nitric Oxide ; therapeutic use ; Random Allocation ; Rats ; Rats, Wistar ; Skin Transplantation ; methods ; Surgical Flaps ; Treatment Outcome

OBJECTIVETo study the relationship between pulmonary surfactant-associated protein B (SP-B) gene polymorphisms and their susceptibility to neonatal respiratory distress syndrome (RDS).

METHODSEighty-eight preterm infants with RDS (RDS group) and 103 infants without RDS (control group) were enrolled. The genomic DNA was isolated using DNA kits. Polymerase chain reaction with restriction fragment length polymorphism technique was used to detect the genotype and allele frequency of the SP-B -18A/C and SP-B 1580C/T single nucleotide polymorphisms. The association between the polymorphisms and RDS was analyzed.

RESULTSSP-B -18A/C and SP-B 1580C/T were found to be polymorphic in both RDS and control groups. The frequencies of CC genotype (X2=12.26, P<0.01) and C allele (X2=11.97, P<0.01) of SP-B 1580C/T were significantly higher in the RDS group than in the control group. The C allele significantly increased the risk of RDS (OR=2.26, 95%CI: 1.42-3.60). The frequencies of genotype and allele of SP-B -18A/C showed no significant difference between the two groups.

CONCLUSIONSSP-B 1580C/T polymorphism contributes to the etiology of RDS and may serve as the susceptibility gene for RDS. The C allele increases the risk of RDS. SP-B -18A/C shows no association with the etiology of RDS.

Genetic Predisposition to Disease ; Genotype ; Humans ; Infant, Newborn ; Polymorphism, Single Nucleotide ; Pulmonary Surfactant-Associated Protein B ; genetics ; Respiratory Distress Syndrome, Newborn ; etiology ; genetics

Objective To discuss the clinical value of transabdominal sonography after bowl preparation in diagnosis of ileocecal valve syndrome ( IVS) .Methods The ultrasonic features of IVS in 37 cases were summerized and correlated with the follow-up findings after conservative treatment or the pathologic results after operation .Twenty-eight cases were confirmed by follow-up and 9 cases by operative pathology.Results Among the 37 cases of IVS,28 were idiopathic IVS (75.7%,28/37) and 9 were secondary IVS (24.3%, 9/37%).For the secondary cases, the primary diseases included 5 acute appendicitis,2 Meckel diverticulum,1 terminal ileitis and 1 carcinoma of ascending colon .The diagnostic accuracy rate of ultrasound was 89.2%(33/37).Misdiagnosis rate was 10.8%(4/37),including 1 case of idiopathic and 3 cases of secondary IVS .The IVS ultrasonic images coulde be displayed clearly using 7.0-10.0 MHz probes.In fasting examination,three ultrasonic characteristic signss were found in interminal ileum region at the right lower abdomen .And these features were bagel-shaped sign [91.9%(34/37),average size (1.9 ±1.6) cm ×(0.8 ±0.3) cm],short sleevelet-shaped sign [91.9% (34/37,average size (2.1 ± 0.4)cm ×(1.3 ±0.2) cm],and rose-shaped sign [83.8% (31/37),average size (1.4 ±0.2) cm × (1.0 ±0.2) cm].The shapes of some signs were changeable when the probe compressed .In the case of idiopathic IVS ,several pathologic changes could be seen on sonography after intestinal tract filling of oral 20%mannitol,including slight thickened mucosa and submucosa of erminalileum ,enlarged ieocecal valve and the crocodile-mouth sign.Conclusions Transabdominal ultrasonic examination with high frequency probe after bowl preparation plays an important role in diagnosis of IVS .The method is simple and accurate and should be recommended and applied clinically .

Objective To investigate the accessibility of HIV-related public health services among cross-border couples living in Dehong Prefecture and age differences. Methods A cross-sectional survey was conducted among cross-border couples in Dehong Prefecture from January 2017 to July 2019. Results In total, 32 182 participants were included. The proportion of people who had received HIV testing services, HIV-related intervention services in past year, care and help in the past year, and participated in new rural cooperative medical services (NCMS) were 57.8%, 92.7%, 6.5% and 94.5%, respectively; and the latter three services were significantly different across age groups (P<0.001). In multivariable Logistic regression model, variables significantly associated with having ever received HIV testing services older age (51-85 years: OR=0.71, 95% CI: 0.63-0.81), women (OR=1.14, 95% CI:1.03-1.25), county/city (Longchuan: OR=6.30, 95% CI: 5.72-6.93; Lianghe: OR=1.27, 95% CI: 1.11-1.44; Yingjiang: OR=0.88, 95% CI: 0.82-0.94), Dai ethnic minority (OR=1.60, 95% CI: 1.50-1.72), marriage registration (marriage registration for border inhabitants: OR=0.60, 95% CI: 0.56-0.65; non-registration: OR=0.66, 95% CI: 0.62-0.70), years of marriage (4-5 years: OR=1.21, 95% CI: 1.12-1.31; 6-60 years: OR=1.30, 95%CI:1.22-1.39), having not received care and help in the past year (OR=0.64, 95% CI: 0.58-0.71) and having not participated in NCMS (OR=0.58, 95% CI: 0.52-0.65). Conclusions The accessibility to HIV-related public health services are relatively high among cross-border couples in Dehong Prefecture. However, the relatively low proportion of receiving AIDS testing services, particularly among certain groups and counties/cities, need to be strengthened.

OBJECTIVE: To observe the effect of daily low-intensity pulsed ultrasound (LIPUs) therapy on improving the enchondral bone formation in lumbar fusion in rabbit models, and to explore its possible mechanism. METHODS: Posterolateral noninstrumented bilateral fusions were performed at the L5 approximately L6 levels in 20 New Zealand rabbits. The autograft iliac bone was implanted on the left side, and the hydroxyapatite bioceramic artificial bone on the right. The rabbits were divided into a treatment group and a control group randomly. One week after the surgery, LIUPs was administered for 20 minutes per day for 4 weeks over the fusion site in the treatment group and false treatment was used in the control group. Post-anterior X-ray photographs were taken to determine the conditions of fusion area, and then, rabbits were killed and the fusion tissues were obtained. Chondrocytes were detected by histological and cytological methods. RESULTS: Compared with the control group, the fusion rate of the treatment group was significantly up-regulated (P<0.05). There was plenty bone trabecula in the fusion area in the treatment group, the number of chondrocytes was also higher than that of the control group (P<0.05), and there was no statistical difference in the number of chondrocytes between the iliac and artificial bone tissues after the treatment(P>0.05). CONCLUSION Low-intensity pulsed ultrasound therapy improves the endochondral bone formation in the lumbar spine fusion in rabbit models.
Animals ; Chondrocytes ; cytology ; Durapatite ; therapeutic use ; Female ; Ilium ; transplantation ; Lumbar Vertebrae ; surgery ; Male ; Osteogenesis ; radiation effects ; Rabbits ; Random Allocation ; Spinal Fusion ; methods ; Ultrasonic Therapy ; methods

BACKGROUNDSeveral studies have revealed that adipose-derived mesenchymal stem cells (ADSCs) can be used as seed cells for the treatment of spinal cord injury (SCI). Chondroitinase ABC (ChABC) decomposes chondroitin sulfate proteoglycans in the glial scar that forms following SCI, allowing stem cells to penetrate through the scar and promote recovery of nerve function. This study aimed to establish ADSCs that stably express ChABC (ChABC-ADSCs) and evaluate the migratory capability of ChABC-ADSCs in vitro.

METHODSADSCs were obtained from Sprague-Dawley rats using secondary collagenase digestion. Their phenotypes were characterized using flow cytometry detection of cell surface antigens and their stem cell properties were confirmed by induction of differentiation. After successful culture, ADSCs were transfected with lentiviral vectors and ChABC-ADSCs were obtained. Proliferation curves of ChABC-ADSCs were determined using the Cell Counting Kit-8 method, ChABC expression was verified using Western blotting, and the migration of ChABC-ADSCs was analyzed using the transwell assay.

RESULTSSecondary collagenase digestion increased the isolation efficiency of primary ADSCs. Following transfection using lentiviral vectors, the proliferation of ChABC-ADSCs was reduced in comparison with control ADSCs at 48 h (P < 0.05). And the level of ChABC expression in the ChABC-ADSC group was significantly higher than that of the ADSC group (P < 0.05). Moreover, ChABC-ADSC migration in matrigel was significantly enhanced in comparison with the control (P < 0.05).

CONCLUSIONSSecondary collagenase digestion can be used to effectively isolate ADSCs. ChABC-ADSCs constructed using lentiviral vector transfection stably express ChABC, and ChABC expression significantly enhances the migratory capacity of ADSCs.

Adipocytes ; cytology ; metabolism ; Adipose Tissue ; cytology ; Animals ; Cell Differentiation ; physiology ; Cell Movement ; physiology ; Cell Proliferation ; physiology ; Cells, Cultured ; Chondrocytes ; cytology ; metabolism ; Chondroitin ABC Lyase ; metabolism ; Flow Cytometry ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Osteoblasts ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley