Effects of RNA M6A demethylase ALKBH5 gene deficiency on morphology and function of cerebellum in aged mice / 中华病理学杂志

Objective: To investigate the effects of RNA m6A demethylase ALKBH5 gene deficiency on cerebellar morphology and function in the aged mice, and to explore the role of ALKBH5 in cerebellar degeneration. Methods: Western blot was performed to detect the protein level of ALKBH5 in the cerebellum of wild-type mice of various ages. The expression of NeuN, Calbindin-D28K, MAP2, GFAP and other proteins in the cerebella of middle-aged (12-month-old) and aged (18-month-old) wild-type mice and ALKBH5-/- mice was examined using immunohistochemistry. The balance beam test and gait analysis were performed to test the balance ability and motor coordination of the mice. Results: With aging of the mice, the expression of ALKBH5 in the cerebellum increased gradually in an age-dependent manner. In the aged mice, but not middle-aged mice, the body weight, whole brain weight and cerebellum weight of ALKBH5-/- mice decreased by 15%, 10% and 21%, respectively (P<0.05). The expression of ALKBH5 in the Purkinje cells was much higher than that in other types of neural cells. Correspondingly, ALKBH5-deficiency caused 40% reduction in the number of Purkinje cells, as well as the length and density of neuronal dendrites in the aged mice (P<0.01). In addition, the time for the aged ALKBH5-/- mice to pass the balance beam was 70% longer than that of the wild type mice of the same age, with unstable gaits (P<0.01). Conclusions: Gene deficiency of RNA m6A demethylase ALKBH5 causes cerebellar atrophy, Purkinje neuron loss and damage in the aged mice. These changes eventually affect mice's motor coordination and balance ability. These results suggest that imbalanced RNA m6A methylation may lead to neurodegenerative lesions in the cerebellum of mice.

Simultaneous quantitative analysis of nine saponins in Gynostemma pentaphyllum before and after heat processing based on UPLC-Q-Trap-MS / 中国中药杂志

Heat-processed Gynostemma pentaphyllum has strong biological activity, and saponins are the main components. To investigate the changes of saponins in G. pentaphyllum before and after heat processing, the present study determined and analyzed the content of nine saponins in G. pentaphyllum from Zhangzhou of Fujian and Jinxiu of Guangxi by ultra-high performance liquid chromatography with quadrupole ion-trap mass spectrometry(UPLC-Q-Trap-MS). The separation of the analytes was performed on an ACQUITY UPLC BEH C_(18) column(2.1 mm×50 mm, 1.7 μm) at 30 ℃, with acetonitrile and 0.1% formic acid in water as the mobile phase by gradient elution, and the flow rate was 0.3 mL·min~(-1). Quantitative analysis was performed using electrospray ionization source(ESI) in the multiple reaction-monitoring(MRM) mode. The results showed that the content of saponins with biological activities increased after heat processing. Specifically, gypenoside L, gypenoside LI, damulin A, damulin B, ginsenoside Rg_3(S), and ginsenoside Rg_3(R) in G. pentaphyllum produced in Zhangzhou of Fujian increased by 7.369, 8.289, 12.155, 7.587, 0.929, and 1.068 μg·g~(-1), respectively, while the content of ginsenoside Rd, gypenoside LVI, and gypenoside XLVI, which were abundant in the raw materials, decreased by 0.779, 19.37, and 9.19 μg·g~(-1), respectively. The content of gypenoside L, gypenoside LI, damulin A, damulin B, ginsenoside Rg_3(S), and ginsenoside Rg_3(R) in G. pentaphyllum produced in Jinxiu of Guangxi increased by 0.100, 0.161, 0.317, 0.228, 3.280, and 3.395 μg·g~(-1), respectively, while the content of ginsenoside Rd, gypenoside LVI, and gypenoside XLVI in the raw materials was reduced by 1.661, 0.014, and 0.010 μg·g~(-1), respectively. The results suggest that heat processing is an effective way to transform rare gypenosides. Furthermore, it is found that there are great differences in the content of gypenosides in different regions.

Dihydroartemisinin Induces AML cell Apoptosis by Inhibition of PTEN/AKT pathway / 中国实验血液学杂志

OBJECTIVE@#To study the effects of dihydroartemisinin (DHA) on the proliferation and apoptosis of acute myeloid leukemia (AML) cells.@*METHODS@#The effects of DHA on the proliferation of acute myeloid leukemia cells and the inhibitory effect of Z-VAD-FMK on the DHA-induced cell apoptosis were detected by CCK-8 assay. The expression level of cleaved-caspased 3 was detected by indirect immunofluorescence. Western blot was used to quantify the protein expression of PTEN, p-Akt, AKT, β-actin, and the apoptosis-associated proteins, such as C-PARP, Cleaved-caspase3 and Caspase3 respectively.@*RESULTS@#DHA induced the AML cell apoptosis with concentration-dependent manner (r=-0.959, r=-0.956). The DHA could induce the accumulation of cleaved-caspase 3 and C-PARP in AML cells, activate PTEN gene and inhibited Akt phosphorylation. Apoptosis inhibitor Z-VAD-FMK could partially restored the activity of DHA-inhibited cell proliferation.@*CONCLUSION@#Dihydroartemisinin induces AML cell apoptosis by inhibition of PTEN/AKT pathway. Dihydroartemisinin is expected to be a safe and effective drug for treatment of acute myeloid leukemia.

Mechanism of Sorafenib and Decitabine Inducing Apoptosis of Diffuse Large B-Cell Lymphoma Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of sorafenib combined with decitabine on the viability and apoptosis of diffuse large B-cell lymphoma cell line OCI-LY1 and its mechanism.@*METHODS@#Sorafenib at 1.5μmol/L or decitabine at 25μmol/L was used to treat the cells alone or in combination. The viability of OCI-LY1 cells was detected by CCK8 assay; the PI positive cells were observed by fluorescence microscopy; the cell proliferation and ROS levels were measured by flow cytometry; The expression levels of proteins related to apoptosis were detected by Western blot.@*RESULTS@#Compared with the control group, treatment with sorafenib and decitabine alone or in combination inhibited the cell proliferation, activated ROS formation and induced apoptosis finally. Sorafenib in combination with decitabine produced a synergistic effect. Western blot analysis showed that sorafenib combined with decitabine significantly up-regulated the levels of Bax/Bcl-2, P53, C-Caspase3 and C-PARP and activated apoptosis by inhibiting PI3K-AKT pathway.@*CONCLUSION@#Sorafenib combined with decitabine induces the apoptosis of diffuse large B-cell lymphoma cell line OCI-LY1 by inhibiting PI3K-AKT pathway and activating P53.

Dihydroartemisinin Induces Apoptosis of Human Acute T Lymphocytic Leukemia Cells by Activating Oxidative Stress / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of dihydroartemisinin (DHA) on the proliferation and apoptosis of human T-cell acute lymphoblastic leukemia (T-ALL) Jurkat cell.@*METHODS@#The effects of DHA on the proliferation of Jurkat cells and the recovery of DHA-inhibited cell viability by N-acetyl-L-cysteine (NAC) were examined by CCK-8 assay. Flow cytometry was performed to analyze the cell apoptosis and generation of reactive oxygen species (ROS). Western-blot was used to detected protein expression of DNA damage-related genes, as well as apoptosis-associated genes, respectively.@*RESULTS@#DHA inhibited the proliferation of Jurkat cells, and shows a concentration-dependent manner(r =0.936), and NAC could partially restore the activity of DHA on cell proliferation inhibition. With the increase of drug concentration, the apoptosis rate (r =0.946) and ROS accumulation was increased (r =0.965). Western blot showed that the protein expressions of DNA damage-related gene γ-H2AX and apoptosis-related genes p53, c-Caspase3, BAX and cPARP were significantly increased, and BCL-2 protein expression was decreased.@*CONCLUSION@#DHA can induce ROS production in Jurkat cells, which can cause DNA damage, activate the P53 apoptotic pathway, and promote apoptosis of cells.

Observation on the efficacy and mechanism of Hirudoid in preventing psoriasis-like dermatitis in mice / 中国医师杂志

Objective To observe the preventive effect of Hirudoid (MPS) ointment on psoriasislike dermatitis in mice and explore its mechanism.Metbods 21 Balb/C mice were randomly divided into three groups:blank control group,positive control group (imiquimod group) and treatment group (imiquimod + Hirudoid group).The blank control group was treated with imiquimod matrix.The positive control group was treated with 5％ imiquimod cream to induce psoriatic dermatitis in mice's back.The treatment group was treated with 5％ imiquimod cream in the morning and Hirudoid cream in the afternoon.The changes of skin erythema,scales,infiltration severity and skin histopathology in the experimental area of mice back and serum levels of interleukin (IL)-17A and protein expression of tumor necrosis factor-α (TNF-α)in the three groups of mice were observed.Results The clinical and pathological findings of skin lesions in the positive control group showed that 5％ imiquimod cream successfully induced psoriasis-like dermatitis in mice.The appearance and pathological feature of skin lesions in the blank control group were almost close to normal skin.The degree of erythema,infiltration and scales in the back of the treatment group was significantly less than that in the positive control group,and the degree of microscopic inflammation in the treatment group was also lighter than that in the positive control group;The serum levels of IL-17A and TNF-α in the treatment group and the blank control group were significantly lower than that of the positive control group,with statistically significant difference (P ＜ 0.05).Conclusions Hirudoid can significantly reduce imiquimod-induced psoriasis-like dermatitis by inhibiting the expression of IL-17A and TNF-α.

Research on competitiveness of traditional Chinese medicine industry's international trade based on diamond theory / 中国中药杂志

China has a long history of the international trade of traditional Chinese medicine(TCM).And the export products mainly composed of Chinese herbal medicine and its extracts with Chinese patent herbal medicine and health care products as complementary items.The international trade of TCM faces problems of structural disequilibrium in export products and trade barriers.In this study,we used Michael Porter's diamond model to analyze the international competitiveness of TCM industry.We found that TCM industry in China was rich in production factors and broad in market demands,but lack of the related and supporting industries.In addition,compared with the herbal medicine manufacturers in European,American and Japanese,enterprises in China were weaker in the strategy making,market positioning and industry competing.The development of the international market of herbal medicine,the arrival of the aging society,and the introduced policies of the TCM,provide great opportunities for TCM industry' s development.In order to improve the competitiveness of the TCM industry,we propose to increase the international recognition of TCM by developing clinical study,cope with international trade barrier by strengthen international standardization research,and improve the competitiveness of TCM industry by economies of scale formed by the accumulation of the pharmaceutical industry.

Research on international competitiveness of ginseng trade / 中草药

To study the data of the trade of the ginseng products in the UN COMTRADE database and analyze the overall situation of the international trade of the ginseng products. China, South Korea, Canada, and the United States are not only the market for the production of ginseng products, but also the main consumer markets for Ginseng products. Hong Kong, China is dominated by entrepot trade and is an important consumer market for ginseng products as well. The four countries mentioned above and Hong Kong, China, play a decisive role in the international trade of ginseng products. Using trade specialization coefficient (TSC), international market share (IMS), revealed comparative advantage (RCA), and competitive advantage (CA), the trade status of the Ginseng products in the four countries from 2007 to 2015 was studied, and the situation of its trade competitiveness was analyzed. The results showed that the average values of three indicators of international trade competitiveness of Canada were ranked the first with the absolute superiority except the mean value of TSC, which was slightly lower than that of South Korea. The average of TSC indicators of South Korea was ranked first, the means of RCA and CA were ranked the second in the world. As for China, the three indexes were all ranked the third except for IMS, ranked the second. However, the four indicators in the United States were all ranked in the last place.

Effects of gravitational traction and targeted regulation of caspase3 on the degenerative intervertebral disc cells in rabbits / 解放军医学杂志

Objective To explore the effects of gravitational traction and target regulation of caspase3 on the degenerative intervertebral disc cells in rabbits.Methods Rabbits nucleus pulposus cells transfected with caspase3 siRNA or negative control siRNA were incubated in serum-starved medium for 6 hours and 48 hours.The expression of caspase3 siRNA and cell apoptosis were analyzed by RT-qPCR and Annexin V-fluorescein staining.In order to create intervertebral disc degeneration model,the right anterior side of the annulus fibrosus of lumbar vertebrae of 35 rabbits were damaged by 16-gauge needle.After confirming the success of modeling,35 animal models were randomized into 3 groups:model group (n=10),negative siRNA group (n=10) and caspase3 siRNA group (n=5).Either negative control siRNA or caspase3 siRNA was injected into the center of nucleus pulposus using a 26-gauge needle from the left anterior side,while the model group received no injection.Nucleus pulposus tissue of 5 rabbits selected randomly from every group after 48 hours were analyzed by PCR and Western blotting for caspase3 mRNA and protein expressions.Half of the casepase3 siRNA group were selected randomly and received a routine gravitational traction using a model of our own design,30min per day for 2 weeks,while other groups received no treatment.TNF-o and IL-1β expression levels and histopathological observations were performed after intervention.Results Expression of caspase3 mRNA in rabbit nucleus pulposus cells transfected with caspase3 siRNA decreased obviously in serum-starved medium,and the apoptosis rate of cells cultured in serum-starved medium decreased significantly (P＜0.05).Caspase3 mRNA and caspase3 protein expression of nucleus pulposus injected by caspase3 siRNA down-regulated in the caspase3 siRNA group.Compared with model group and negative control siRNA group,TNF-α and IL-1β expression levels of caspase3 siRNA traction group decreased significantly (P＜0.05),but there was no statistical significance compared with caspase3 siRNA group (P＞0.05).Pathological observation revealed that viable cell number and extracellular matrix contents increased and collagenous fibers arranged regularly in caspase3 siRNA traction group.Conclusion Gravitational traction and target regulation of caspase3 can prevent apoptotic cell death and delay early intervertebral disc degeneration.

Research on ginseng trade competitiveness between China and Korea / 中国中药杂志

Ginseng is one of China's valuable Chinese herbal medicines, with a long using history. Ginseng has worldwide reputation, and widely used in food, medicine, health products, cosmetics and other production. China and South Korea have a big ginseng industrial, and sharing half of the export market. The ginseng export competitiveness analysis seems important and necessary between China and South Korea. In this paper, the data of customs and trade of ginseng in COMTRADE database were studied, and ginseng export competitiveness was analyzed between China and Korea. The results showed that the ginseng export competitiveness of Korean more competitive than China. Contrast with China, South Korea using only 15% total amount of ginseng exports and produced the same total export amount. This article has the reference value to the traditional Chinese medicine resources management and the economics research. On this basis, this paper further discusses the problems that should be paid attention to in the development of ginseng industry in China.

Effect of Ultra-early Hyperbaric Oxygenation on Bone Calcium, Biomechanical Properties and Bone Collagen of Femur in Rats with Complete Spinal Cord Transection Injury / 中国康复理论与实践

Objective To explore the effect of ultra-early hyperbaric oxygenation (HBO) on bone calcium, biomechanical properties and bone collagen of femur in rats with complete spinal cord transaction. Methods A total of 75 Sprague-Dawley rats were randomly divided in-to sham group (n=15), model group (n=20) and HBO group (n=40). HBO group was divided into three hours group (HBO1 group, n=20) and twelve hours group (HBO2 group, n=20). All groups underwent laminectomy at T10, while the model group, HBO1 group and HBO2 group underwent complete spinal cord transection at the same level. Three hours and twelve hours after surgery, HBO1 group and HBO2 group received HBO, respectively, for three courses with ten days in a course. After treatment, the femoral biomechanical properties, bone calcium and hydroxyproline (Hyp) were determined. The morphology of bone trabecula and the bone collagen was observed with HE stain-ing and Masson triad color staining, respectively. Results After treatment, compared with the sham group, the femoral biomechanical proper-ties, the content of bone calcium and Hyp decreased in the model group (P<0.05);compared with the model group and HBO2 group, they in-creased in HBO1 group (P<0.05). The number of bone trabecula and the bone collagen decreased, and derangement and sparseness were ob-served in the model group;however, the changes were substantially mild in HBO1 group. Conclusion Ultra-early HBO could increase the content of bone calcium and Hyp of femur, improve the morphology of the femur bone collagen, and improve the femoral biomechanical properties in rats with complete spinal cord transection.

Improved methods of establishing rabbit VX2 hepatocarcinoma models / 中国组织工程研究

BACKGROUND:Rabbit VX2 hepatocarcinoma models are characterized by expanded and infiltrative growth, abundant blood supply, similar pathological and imaging features to human hepatocelular carcinoma, so this model is ideal for imaging experiments. OBJECTIVE: To establish rabbit VX2 hepatocarcinoma models and compare two different laparotomy methods in establishing rabbit VX2 hepatocarcinoma models. METHODS:Thirty New Zealand rabbits were randomly divided into two groups: traditional laparotomy (tumor particles were directly embedded into the liver with tweezers) and improved laparotomy (tumor particles were injected into the liver through a 15G syringe needle). The growth features of the tumor established in two different ways were compared by ultrasound. RESULTS AND CONCLUSION:The tumor formation rate was 100% in the two groups; the single rate was respectively 50% and 90%; the ectopic implantation rate was 50% and 10%, there were significant differences between two groups (P < 0.05). The mortality rate within 2 weeks was 33% and 0 in the two groups, with significant differences (P < 0.05). The improved laparotomy method is feasible for modeling rabbit VX2 hepatocarcinoma, due to minimal injury, short operation time, high success rate and stable features.

Perioperative nursing of esophageal cancer patients undergoing chest laparoscopic intrathoracic esophagus anastomosis / 现代临床护理

ObjectiveTo summarize the experiences for nursing esophageal cancer patients undergoing chest laparoscopic intrathoracic esophagus anastomosis.Method The clinical data of 67 esophageal cancer patients before and after chest laparoscopic intrathoracic esophagus anastomosis were retrospectively reviewed for summarizing nursing experience.Result All 67 patients were discharged successfully,including 2 contracting anastomotic leak symptoms but cured by conservative therapy,4 contracting lung infections but cured after antimicrobial therapy,and another 2 contracting acute renal failure and cured by way conservative therapy as well.Conclusion Comprehensive preoperative preparation,accurate surgical operation and effective pre- and post-operative nursing measures can accelerate the healing,improve the cure rate and reduce the occurrence of complications.

Growth, reproduction and blood physiological and biochemical indexes in an atopic dermatitis mouse model / 中国组织工程研究

BACKGROUND:The skin lesions, pathological and immunological characteristics of Nc/Nga mice are consistent with human atopic dermatitis, and as an atopic dermatitis animal model, it has great research value. But there are no reports on the physiological and biochemical parameters of Nc/Nga mice in China. OBJECTIVE:To observe the growth and reproduction and blood physiological and biochemical parameters of atopic dermatitis model of NC/Nga mice METHODS:The data of reproductive performance of NC/Nga mice from the first to the third generation was analyzed, including mean litter size, weaning rate, pregnancy rate and generation interval. The body mass of 60 Nc/Nga mice with 1-56 days old was measured, 30 mice of female and male, then the growth curve was draw. The blood samples from the infraorbital vessels were col ected to detect the blood physiological and biochemical parameters in the mice. RESULTS AND CONCLUSION:The mean delivery interval of Nc/Nga mice was (25.8±3.1) days, mean litter size was (7.5±2.5) mice, mean weaning rate was (97.2±1.2)%, the mean pregnancy rate was (97.0±1.4)%, and there was no significant difference among the three generations of these mice (P>0.05). The body mass of Nc/Nga mice was increased with the time increasing of days, the body mass of the mice was maximal within 1-2 weeks post-weaning, and there was no significant difference in body mass between males and females at 6 weeks post-weaning (P>0.05). Comparison of the blood physiological and biochemical parameters of mice at the same age between males and females showed that the levels of red blood cells and hemoglobin in female mice were significantly higher than those in the male mice;the platelet count in the male mice was significantly higher than that in the female mice (P<0.05);the triglycerides and albumin levels in the female mice were higher than those in the male mice (P<0.05). The results indicate that the gender and age may influence the blood physiological and biochemical parameters of Nc/Nga mice.

Study of RON mediated invasion of Raji cell line and drug-target effects / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the proto-oncogene RON mediated aggression of Raji cells and the inhibitory effects by monoclonal antibody Zt/f2 (2f2).</p><p><b>METHODS</b>The effects of RON ligand macrophage stimulating protein (MSP) (2.0 nmol/L) and inhibitory Zt/f2 (2F2) (2.0 nmol/L) antibody on proliferation of RON positive Raji cells after treatment for 24 and72 hours were detected by MTT method, colony formation units (CFU) of Raji cells by methylcellulose semi solid culture, Raji cells apoptosis and cell cycle analysis by AnnexinV/PI double staining, expression of RON, apoptosis-related proteins, and cyclins by Western blot.</p><p><b>RESULTS</b>(1)Compared with the cell viability (1.0) and counts of CFU (103.6±7.0) in control group, Raji cells after MSP treatment had better viability (1.35±0.20) and CFU counts (133.7±10.4) (P<0.05), but worse viability (0.68±0.11) and CFU counts (66.3±6.1) after Zt/f2 (2F2) treatment (P<0.05). (2)Percentage of Raji cells apoptosis after Zt/f2 (2F2) antibody treatment (12.16±2.33)% was significantly increased than the control (2.89±1.03)% (P<0.05). The percentage of Raji cells arrested in G0/G1 phase was increased after Zt/f2 (2F2) antibody treatment as compared to the control [ (54.96 ±3.70)% vs (39.10±2.30)%, (P<0.05) ]. (3) High-level of RON phosphorylation and β-catenin expression activated by MSP could be inhibited significantly by Zt/f2 (2F2), which also up-regulated the expression of caspase-3, caspase-8, caspase-9 and PARP and down-regulated anti-apoptotic MCL-1 gene and inhibitor of apoptosis protein XIAP expression, accompanied with G1 phase protein changes accordingly.</p><p><b>CONCLUSION</b>MSP could aggravate Raji cells proliferation. Inversely, Zt/f2 (2F2) could inhibit proliferation and induce apoptosis by inhibition of RON phosphorylation and up-regulation of apoptosis related proteins.</p>

Effects of rhubarb navel therapy on the functions of colostomy in rectal cancer patients with colostomy / 中国实用护理杂志

Objective To explore the effects of rhubarb navel therapy on fecal continence in rectal cancer patients with colostomy.Methods 60 patients with colostomy were randomly divided into the experimental group and the control group with 30 patients in each group.Patients in the control group received routine therapy and health education whereas the experimental group received 30-day extra rhubarb navel therapy started on the 15th day after the operation.The two groups were compared in postoperative 3 months with the following factors:the functions of colostomy and the formation of bowel regularity.Results The scores of the scale about the functions of permanent abdominal colostomy were improved in both groups.However,the score in the experimental group was much higher than that in the control group.The number of patients forming bowel regularity in the experimental group was more than that in the control group (19 and 4 cases respectively).Conclusions Rhubarb navel therapy is beneficial to rectal cancer patients with colostomy to improve the functions of colostomy and form the bowel regularity.

Comparison of two rat models of myocardial infarction / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare two rat models of myocardial infarction established by coronary artery ligation and harmonic scalpel method for their advantages, disadvantages and applications.</p><p><b>METHODS</b>Sixty Sprague Dawley rats were randomly divided into sham-operated group, coronary artery ligation group and harmonic scalpel group. In the latter two groups, myocardial infarction was induced by ligation of the left anterior descending coronary artery and coagulation of the artery, respectively. Four weeks after the operations, the rats were sacrificed to measure the serum levels of CK, CK-MB, LDH and AST, and the hearts were collected for histopathological observation with HE staining.</p><p><b>RESULTS</b>The rats in the sham-operated group and coronary artery ligation group showed no significant differences in serum levels of CK , CK-MB , LDH or AST. Histologically, the myocardial tissues of the two models all showed coagulation necrosis and inflammatory cell infiltration, with granulation tissue ingrowth in the infracted tissues and scar formation. The mortality rate of the rats in coronary artery ligation group was 50% at 4 weeks, significantly higher than that of the harmonic scalpel group (20%, P<0.05).</p><p><b>CONCLUSION</b>Coronary artery ligation and harmonic scalpel method are both effective means to produce rat models of myocardial infarction. The latter method can establish a more stable model for studying the mechanism and treatment of myocardial infarction.</p>

The establishment of PCR system to identify Bungarus multicinctus rapidly / 药学学报

The purpose of the present study is to establish a rapid and effective PCR method for the identification of B. multicinctus. Based on sequence alignment of B. multicinctus and its adulterants, we found that Cyt b gene is a good molecular genetic marker for the authentication of B. multicinctus. On the basis of the sequence data, a pair of highly specialized primers was designed. The templates were extracted by the DNA purification system. Key factors such as annealing temperature, concentration of Taq enzyme and cycle numbers were analyzed and optimized. The modified PCR program consisted of an initial denaturation step at 95 degrees C for 5 min, followed by 30 cycles of 95 degrees C for 30 s and 55 degrees C for 45 s and a final extension at 72 degrees C for 5 min. Thirteen samples of B. multicinctus were identified accurately from their 20 adulterants in 4 hours. The results indicated it is a highly accurate, rapid and applicable method for the authentication of B. multicinctus.

Overexpression of connexin 45 in rat mesenchymal stem cells improves the function as cardiac biological pacemakers / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Extensive research toward creating a biological pacemaker by enhancement of inward depolarizing current has been performed. However, studies have mainly focused on inducing spontaneous activity and have not adequately addressed ways to improve pacemaker function. In this study we attempted to improve pacemaker function by altering connexin expression in rat mesenchymal stem cells (MSCs) to a phenotype similar to native sinus node pacemaker cells.</p><p><b>METHODS</b>To generate a biological pacemaker, MSCs were transduced with a cardiac pacemaker gene-hyperpolarization-activated cyclic nucleotide-gated channel 4 (HCN4), via transfection with a lentiviral vector. Funny current (I(f)) in HCN4(+) MSCs was recorded by voltage-clamp. Overexpression of connexin 45 (gene Gja7) in MSCs was achieved by transfection with the plasmid pDsRED2-N1-Gja7-RFP. Double-immunolabelling with anti-connexin 43 and anti-connexin 45 antibodies were used to identify the gap junction channels. The effects of the genetically modified MSCs on cardiomyocyte excitability were determined in MSCs cocultured with neonatal rat ventricular myocytes. Spontaneous action potentials of neonatal rat ventricular myocytes were recorded by current-clamp.</p><p><b>RESULTS</b>High level time- and voltage-dependent inward hyperpolarization current that was sensitive to 4 mmol/L Cs(+) was detected in HCN4(+) MSCs, confirming that HCN4 acted as I(f) channels in MSCs. Connexin 43 and connexin 45 were simultaneously detected in CX45(+) MSCs. Beating frequency was (82 +/- 8) beats per minute (n = 5) in myocytes cocultured with non-transfected control MSCs, versus (129 +/- 11) beats per minute (n = 5) in myocytes cocultured with HCN4(+) MSCs. Myocytes cocultured with MSCs cotransfected with HCN4 and connexin 45 had the highest beating frequency at (147 +/- 9) beats per minute (n = 5).</p><p><b>CONCLUSION</b>These findings demonstrate that overexpression of connexin 45 and subsequent formation of heteromeric connexin 45/connexin 43 gap junction channels in HCN4 expressing MSCs can improve their function as cardiac biological pacemakers in vitro.</p>

Effect of ultra-early hyperbaric oxygenation on spinal edema and hind limb motor function in rats with complete spinal cord transection / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the effect of ultra-early hyperbaric oxygenation on spinal edema and hind limb motor function in rats with complete spinal cord transection.</p><p><b>METHODS</b>Fifty-five healthy 3-month-old female SD rats were randomly divided into sham-operated group (n=15), complete spinal cord transection group (CSCT group, n=20) and hyperbaric oxygen group (HBO group, n=20). The rats in the sham-operated group underwent only laminectomy, while those in the other 2 groups underwent CSCT at the T10 level. The rats in HBO group were placed in an oxygen chamber 3 h after the operation for 10 days as a treatment course, and 3 treatment courses were administered at the interval of 6 days. In the first treatment course, 2 hyperbaric oxygenation sessions were given daily, and in the following 2 course, only 1 session was given daily. The recovery of hindlimb motor function was evaluated using the open-field Basso-Beattie-Bresnahan (BBB) scoring system once a week for 6 weeks. All the rats were sacrificed 6 weeks after the operation to measure the water content in the injured tissues.</p><p><b>RESULTS</b>The BBB scores of CSCT group and HBO group gradually increased with the passage of time after the operation, and from week 2 to week 6, HBO group had significantly higher scores than CSCT group (P<0.05). The water content was markedly increased in CSCT group at 6 weeks after the operation as compared with that in the sham-operated group (P<0.01), and significantly reduced in HBO group in comparison with that in the CSCT group (P<0.05).</p><p><b>CONCLUSION</b>Ultra-early HBO can suppress spinal cord edema and promote hindlimb locomotor recovery in rats with complete spinal cord transection.</p>