AIM:To investigate the role of nerve growth factor (NGF) and leukemia inhibitory factor (LIF) in the expression of IL-4 and IL-5 in the splenic lymphocytes of asthmatic rats. METHODS:Sixteen Sprague-Dawley rats were randomly divided into two groups:control group (n = 8) and asthma group (n = 8). The asthmatic model was set up by intraperitoneal injection of 100 mg ovalbumin and 100 mg aluminum hydroxide,and receiving nebulization aspiration of 1% ovalbumin solution 2 weeks later. The splenic lymphocytes from the control rats and asthmatic rats were isolated and cultured with exogenous NGF or LIF at different doses. The mRNA expression of IL-4 and IL-5 in the splenic lymphocytes was respectively detected by reverse transcription-polymerase chain reaction (RT-PCR). The secretions of IL-4 and IL-5 were respectively detected by ELISA. RESULTS:Compared to control group,the expression of IL-4 and IL -5 (P

To investigate the expression of polyadenosine diphospho-ribose polymerase 1 (PARP-1) and p16/ retinoblastoma (Rb) protein in hydroquinone (HQ)-induced TK6 cells and their regulatory mechanisms. Methods According to the 2×2 factorial design model, TK6 cells were divided into PBS-TK6 group and HQ-TK6 group based on HQ exposure, and then sub-divided into non-DOX intervention subgroup and DOX intervention subgroup based on DOX intervention, a total of four groups. The PBS-TK6 group was treated with phosphate buffer saline, and the HQ-TK6 group was treated with HQ at a final concentration of 20.0 μmol/L. The non-DOX intervention subgroup was added with 0.05% dimethyl sulfoxide; and the DOX intervention subgroup was added with PARP-1 agonist DOX at a final concentration of 0.5 μmol/L. The distribution of cell cycle was detected by flow cytometry. The protein expression of p16/Rb, cyclin D1 (cyclinD1), multifunctional protein E2 transcription factor 1 (E2F1), Rb, and p-Rb were detected by Western blot, and the level of p16 ribosylation was detected by immunofluorescence and immunoprecipitation. Results Compared with the PBS-TK6 group, the cell cycle distribution percentage in G0/G1 phase and the relative expression levels of p16 proteins were decreased in the cells of the HQ-TK6 group (all P<0.05). The cell cycle distribution percentage in S phase and the relative expression levels of cyclinD1 and p-Rb proteins were up-regulated (all P<0.05). Compared with the non-DOX intervention group, the cell cycle distribution percentage in G0/G1 and G2/M phases and the relative expression level of p16 protein increased in the DOX intervention group (all P<0.05). The percentage of cells in S phase and the relative expression levels of cyclinD1 and p-Rb proteins were down-regulated (all P< 0.05). The results of interaction effect analysis showed that compared with the non-DOX PBS-TK6 cells, the relative expression levels of Rb and E2F1 protein in the DOX PBS-TK6 cells intervention group were down-regulated (all P<0.05). The relative expression level of Rb protein in non-DOX HQ-TK6 cell group was down-regulated (P<0.05), and the relative expression of E2F1 protein was up-regulated (P<0.05). Compared with DOX PBS-TK6 cell group, the relative expression level of Rb protein in DOX HQ-TK6 cell group was down-regulated and that of E2F1 protein was up-regulated (all P<0.05). Compared with the non-DOX HQ-TK6 cell group, the relative expression level of Rb protein in the DOX HQ-TK6 cell group was up-regulated and that of E2F1 protein was down-regulated (all P<0.05). Conclusion PARP-1 participates in cell cycle regulation by regulating the p16/Rb signaling pathway in TK6 cells.

Isoprene is an important precursor of synthetic rubber material. In our previous study, metabolic engineered Escherichia coli strain (BW-01) was constructed and used to produce isoprene. Based on the theory of protein budget, using synthetic biology strategies including the increased copy number of genes and rare codons, we regulated the expression of key enzyme to improve isoprene production in Escherichia coli strain. Under shake-flask conditions, isoprene productivity of the engineered strain (BW-07) increased by 73% compared with BW-01, reached 761.1 mg/L. It provides a reference for further studies.
Butadienes ; Escherichia coli ; genetics ; metabolism ; Gene Dosage ; Hemiterpenes ; biosynthesis ; Industrial Microbiology ; Metabolic Engineering ; Mevalonic Acid ; Pentanes ; Synthetic Biology

Objective To study the therapeutic effects and related mechanism of Tongmai Yizhi granules on vascular dementia in rats. Methods Bilateral carotid artery ligation was used to establish rat model of vascular dementia (VD). The rats were randomly divided into 6 groups: sham operated,the model control,piracetam (0. 375 g·kg-1 ·d-1 ) as positive con-trol,low dose (2. 5 g·kg-1 ·d-1 ),middle dose(5. 0 g·kg-1 ·d-1 ),and high dose (10. 0 g·kg-1 ·d-1 ) of Tongmai Yizhi granules. Each group was intragastrically administered with 10 mL·kg-1 of corresponding medications for 31 days after the VD model was established. The sham group was given with 0. 9% NaCl solution. Y-electric maze was used to test the learning and memory function of rats at the second and fourth weeks. On the day 32,hippocampal tissues were collected for pathological analy-sis by microscope. The activities of SOD and content of MDA in serum and tissue homogenate were tested. Results Compared with model control group,each dose of Tongmai Yizhi granules obviously shortened the incubation period of VD rats in electric maze test,and accuracy of learning and memory was improved (P<0. 05 or P<0. 01). The activity of SOD was decreased,and the content of MDA was increased after treatment with Tongmai Yizhi granules (P < 0. 01). Hippocampal neuronal cells were disar-ranged,and the number of cone cells was decreased significantly in the model control group. In contrast,the necrosis and degener-ation of hippocampal neurons were alleviated in all Tongmai Yizhi treatment groups. Conclusion Tongmai Yizhi granule can apparently improve learning and memory function of VD rats. It can also improve the activity of SOD and reduce the content of MDA in serum and hippocampus,effectively remove free radicals,and alleviate the injury of hippocampal pyramidal cells.

BACKGROUND: Both biological and physical methods contribute to the repair of white meniscal tears, so the combined use will be better. OBJECTIVE: To study the influence of radiofrequency combined with platelet-rich plasma (PRP) on the repair of white meniscal tears. METHODS: Forty-eight adult New Zealand white rabbits were randomly divided into four groups, including simple suture, radiofrequency, PRP, and combination groups, and then the model of medial meniscus posterior root tears was established in all experimental animals. The simple suture group underwent interrupted mattress suture; the radiofrequency group was treated with radiofrequency (20 W, 45 ℃) after suture; the PRP group received the intra-articular injection of PRP after suture; the combination group was firstly treated with radiofrequency after suture, and then underwent the intra-articular injection of PRP. The gross and histological changes of the meniscus were observed, and the expression levels of transforming growth factor β1, vascular endothelial growth factor and platelet-derived growth factor were detected at 3 and 12 weeks postoperatively. RESULTS AND CONCLUSION: At 12 weeks after operation, the simple suture group showed no healing; the radiofrequency and PRP groups healed partially; and the combination group healed completely, and chondrocytes and collagen fibers arranged regularly. There was a significant difference in the healing rate between combination and simple suture groups (P=0.003). At 3 weeks postoperatively, the expression levels of vascular endothelial growth factor in the radiofrequency, PRP and combination groups were obviously higher than that in the simple suture group, which decreased markedly at the 12th week. At 3 weeks postoperatively, the expression level of platelet-derived growth factor was increased in all groups, especially in the PRP and combination groups. The PRP group showed the highest level of transforming growth factor β1 at 12 weeks postoperatively. These results manifest that the combination of radiofrequency and PRP promotes the repair of white meniscal tears by increasing the cell proliferation, cell mitosis and angiogenesis.

Objective To investigate the corresponds between NF-κB p65 activation and k-ras mutations.Methods NF-κB p65 activation was analyzed by immunochemistry in 167 colorectal cancer specimens in which the k-ras mutation status was confirmed.Resluts Among 167 colorectal cancer specimens screened,59 (35.3 ％) had k-ras mutations and 63 (37.7 ％) had NF-κB p65 activation.k-ras mutations and NF-κB p65 nuclear expression were no significant difference in different sex,age,ECOG score,pathological types,degrees of pathological differentiation,TNM stage,and metastases on lymph nodes (P＞ 0.05).The positive nuclear expression rate of NF-κB p65 was 50.8 ％ (30/59) in specimens with k-ras mutations and 30.6 ％ (33/108) in specimens with wild type k-ras.There was obviously statistical difference between them (P =0.010).Conclusion NF-κB p65 activation in coloretal cancer was associated with k-ras mutations.

Objective To study the feasibility and security about misoprostol combined with fentanyl and propofol applied in visualization of artificial abortion.Methods 1270 cases of pregnant women who requested termination of pregnancy and voluntarily accepted abortion were selected.In the observe group,634 cases of pregnant women received oral misoprostol about 600 mg 90 minutes before surgery.After anesthetized by fentanyl and propofol,painless artificial abortion using visual abortion instrument was carried out.In the control group,636 cases of pregnant women anesthetized by fentanyl and propofol via the venous were carried out artificial abortion.The degree of cervical relaxation,loss of blood,total dose of propofol,surgical time,recovery time,the incidence of abortion complications and so on were observed.Results In the observation group and the control group,the average dosage of propofol are( 14.15 ± 2.37) ml and (16.90 ± 4.52) ml respectively;The average time required for surgery are ( 146.1 ± 38.2) s and ( 199.7 ± 37.3 ) s respectively;the average time required for awakening are ( 6.01 ± 1.90 ) min and ( 8.81 ± 2.02 )in;In the observation group,there are 405 ( 63.9% ) cases of cervical laxity markedly,in the control group,including 145 ( 22.8% ) cases of cervical laxity markedly,there is statistical significance ( P ＜ 0.05 ) between the observation group and the control group.There are no abortion syndrome in both groups.Conclusion Misoprostol combined with fentanyl and propofol was the first choice for painless artificial abortion using ultrasound visualization.

Objective To treat femur fracture in neonates using a new homemade harness designed and observe the effec of this harness. Methods Designed a new harness used in femur fracture in neonates,and used this harness in 7(8 femur fracture)csses,including 6 boys(7 femur fracture)and 1 girl,the age from 1 day to 12 days(average 4.7 days).Those cases included proximal thirds(2 cases)and middle thirds(6 cases)of femur fracture.The angle of fracture was from 44°to 83°(average 62.4°)before treatment. Results The angle of fracture was from 0°to 22°(average 14.0°)after treatment using the harness.Hespitalization was from 2 to 3 days.There were no skin sloughing or harness breaking off.All cases were followed up 6 to 36 months(average 21.3 months).All femur fracture healed in good alignment with leg-length discrepancy＜1 cm.Movement of hip and knee was normal. Conclusions The harness is a better method to treat femur fracture of the proximal and middle thirds in neonates.Advantages of it include simply operation,minimal hospitalization,minimal cost,and easy nursing.

Objective To compare the advantages and disadvantages of the amplification refractory mutation system -polymerase chain reaction (ARMS-PCR) with restriction fragment length polymorphism polymerase chain reaction(RFLP-PCR) technology, then explore its application in detection and analysis of single nucleotide polymorphism (SNP). Methods By using ARMS-PCR and RFLP-PCR the polymorphisms of the MMP-2 in cervical cancer and the normal-control group were analyzed. The research sites were-1306 C/T.The final result was verified by DNA sequencing. Results The results of amplified used ARMS-PCR showed that the genotypes of the SNP site in MMP-2 gene promoter were CC, CT, TT, and the DNA fragments were corresponding with the results of DNA sequencing. Conclusion ARMS-PCR technology has the advantages of lower cost time-saving simple operation and reliable results, while has a high value for research of SNP.

lso.This result suggested that Th17 subset is differentiated in chronic stage of viral myocarditis.