Aim The purpose of this study was to compare the effect of AA、15-HETE、15-KETE and 8(S),15(S)-DiHETE on hypoxic and normoxic rabbit pulmonary arterial rings and to discuss their roles in the formation of PA hypertension.Methods Twelve neonatal rabbits were randomly divided into two groups(n=6):the normoxic group breathing fresh gas(FiO_2 =21%) and the hypoxic group breathing hypoxic gas(FiO_2=10%).After 9 days,they were anesthetized with pentobarbital sodium,and the chest was opened for removal of the heart and lungs en bloc,then PA rings(0.5~1.5 mm in diameter and 3 mm length) were prepared.We used tension studies of PA rings to observe the effect of AA、15-HETE、15-KETE and 8(S),15(S)-DiHETE on hypoxic and normoxic rabbit pulmonary arterial rings.Results ① AA、15-HETE、15-KETE and 8(S),15(S)-DiHETE constricted normoxic rabbit PA rings in a dose-dependent manner,respectively.Constriction of normoxic rabbit PA rings induced by 15-KETE、8(S),15(S)-DiHETE was significant,but that of AA、15-HETE wasn′t significant.② AA、15-HETE、15-KETE and 8(S),15(S)-DiHETE increased hypoxic rabbit PA rings' tension.Constriction of AA、15-HETE on rabbit PA rings in hypoxic group was significantly greater than that of normoxic group(P0.05).③ Constriction of 15-KETE on rabbit PA rings in normoxic group was significantly greater than that of hypoxia group(P

A novel biosensor for aflatoxin B1 detecting has been reported. The biosensor electrode for AFB1 detecting was assembled by immobilized aflatoxin-oxidoreductase using open-ended multi-walled carbon nanotubes as matrix. Its linear range was between 0.16μM and 3.2μM. And if the specific anti-aflatoxin B1 antibody and aflatoxin oxidoreductase were both immobilized on the electrode with Multi-Walled carbon nanotubes, the detection limit of the modified electrode could be 16 nM with a 10 times improved sensitivity. The aflatoxin enzyme biosensor assembled this way strode one step forward its practical application.

AIM:Increasing evidence suggests that carbohydrate-binding proteins play an essential role in tumor growth and metastasis .Ga-lectin-3, a multifunctional protein of an expanding family of β-galactoside-binding animal lectins , is the major nonintegrin cellular laminin-binding protein , and is implicated in a variety of biologic events , such as inflammation and angiogenesis .Because galectin-3 expression was shown to participate in mediating tumor angiogenesis and initiate signaling cascades in several diseases .We hypothe-sized that galectin-3 may promote pulmonary vascular endothelial neovascularization .METHODS:Hypoxic and MCT rat model of pul-monary artery remodeling was used .The mRNA and protein levels of galectin-3 in rats were measured by in situ hybrization and West-ern blot analysis.Endothelial cell (EC) proliferation, migration and tube formation were measured using MTT , cell scratch and Matri-gel assays, respectively.Protein expression was quantitated by Western blot analysis .LC 3A/B staining was detected with cellular im-munofluorescence staining .RESULTS:We found that galectin-3 was localized on the intima and adventitial wall .Galectin-3 was in-creased after rat hypoxia and MCT administration .Galectin-3 promoted EC proliferation , migration and tube formation , while its roles were reversed by RNA interference.Galectin-3 induced Atg 5, Beclin-1, LAMP-2, and LC 3A/B expression increases.Galectin-3 al-so increased LC 3A/B staining in ECs.Akt/mTOR and GSK-3βsignaling pathways were activated after galectin-3 treated ECs using its specific phosphorylation antibodies , while blocked it with LY294002 inhibited cell autophagy and EC dynamic alterations induced by galectin-3.CONCLUSION:These findings demonstrate that galectin-3 can induce an Akt signaling cascade leading to cell autoph-agy, and then the differentiation and angiogenesis of pulmonary artery endothelial cells .

Aim To investigate the effect of 15-ketoeicosatetraenoic acid(15-KETE)and its mechanism through ion channels on rat isolated pulmonary arterial rings by using organ bath technique.Methods Sixteen healthy Wistar rats weighing 220?20 g were divided into two groups(n=8): normoxia group breathing fresh air(FiO_2=21%) and hypoxia group breathing hypoxic air(FiO_2=10%) in a hypoxic box.Pulmonary arteries(PA)were extracted after 9 d and cut into rings(0.5~1.0 mm in diameter and 3 mm in length) for organ bath experiments.Results(1) With increasing concentration from 0 to10~(-6) mol?L~(-1),15-KETE increased PA rings tension gradually in a dosedependent fashion;(2) 4-aminopyridine(2 mmol?L~(-1)),a Kv channel blocker significantly decreased constriction of rat isolated PA rings induced by 15-KETE,and results were similar in two groups;(3) The K_(ATP) channel blocker glyburide(10~(-6)6 mol?L~(-1)) and the BK_(Ca) channel blocker tetraethylammonium(10 mmol?L~(-1)) did not affect constriction of rat isolated PA rings induced by 15-KETE;(4) The BKCa channel blocker nifedipine(10~(-6) mol?L~(-1)) and Ca~(2+)-free Krebs solution significantly decreased constriction of rat isolated PA rings induced by 15-KETE.Conclusion Kv channels play a role in constriction of PA induced by 15-KETE;L-type Ca~(2+) channel blocker and extracellular calcium ion also influence constriction of isolated PA rings induced by 15-KETE.

Aim To test the contribution of 15-HETE on expression of KV1.5 channel under hypoxia condition,using CDC or NDGA to block 15-LO/15-HETE,and to observe the effect of hypoxia on KV1.5 channel protein,mRNA expressions in cultured rat pulmonary arterial smooth muscle cells(PASMCs)and pulmonary arterials(PAs).Methods Western blot,RT-PCR and 15-LO blockers,cinnamyl 3,4-dihydroxy-[alpha]-cyanocinnamate(CDC)or nordihydroguiairetic acid(NDGA)were used to identify the role of endogenous 15-HETE on expression of KV1.5 channel in cultured rat pulmonary arterial smooth muscle cells(PASMCs)and PAs.Results(1)The expressions of KV1.5 channel protein and mRNA in PASMCs and PAs preteated with CDC or NDGA greatly increased than those of PASMCs under hypoxia group.(2)Exogenous 15-HETE added to PASMCs pretreated with CDC or NDGA greatly decreased the expression of KV1.5 than that of adding PASMCs pretreated with CDC or NDGA under hypoxia condition.Conclusion The down-regulation of KV1.5 channel expression caused by hypoxia is through endogenous 15-HETE.

Aim To investigate the effect of SA on induction of ERK1/2 activity in rat pulmonary smooth muscle cells(PASMCs).Methods Western blot analysis was employed to identify the activation of ERK1/2 stimulated by SA at different time points and concentrations in cultured rat PASMCs.Results An unexpected observation showed that ERK1/2 phosphorylation was seen after treatment of SA for 2h at a high concentration(30 ?mol?L-1) but not at lower concentration(from 1 nmol?L-1 to 1 ?mol?L-1).Activation of ERK1/2 pathway could be inhibited by an ERK1/2 inhibitor PD98059 or a protein kinase A(PKA) activator isoproterenol.Conclusion Together,these results suggest that SA has a strong dual regulating effect upon ERK1/2 through PKC and/or PKA pathways in rat PASMCs.

Objective:To investigate the factors related to recurrence and prognosis of retroperitoneal liposarcoma.Method:The clinical data of patients with primary retroperitoneal liposarcoma who underwent surgical treatment in the First Affiliated Hospital of Zhengzhou University from June 2011 to January 2020 were analyzed retrospectively. There were 42 males and 47 females and patients’median age was 53 (26-78). Sixty-five cases were treated by operation in our hospital, and 24 cases were primarily treated by the operation in another hospital. The clinical manifestations of the initial diagnosis included retroperitoneal mass in 41 cases, abdominal distension in 12 cases, abdominal pain in 10 cases, fever in 11 cases, nausea, vomiting and poor appetite in 8 cases, frequent urination and dysuria in 6 cases, and bilateral lower limb edema in 1 case. Preoperative CT imaging showed that the tumor body was located in the retroperitoneal kidney area in 58 cases, while in the retroperitoneal space or the pelvic extraperitoneal space in 31 cases. There were 55 single cases and 34 multiple cases. The median tumor length was 20(3-52) cm. Among the primarily treated 65 patients, 47(72.3%) were considered as primary retroperitoneal liposarcoma by preoperative imaging examination. Among the 89 patients treated by surgery, 78 underwent endoscopic surgery, among which 21 underwent laparoscopic surgery, 38 cases of retroperitoneal laparoscopic surgery, 19 cases of Da Vinci robot-assisted laparoscopic surgery. Open operation was performed in 11 cases. There were 87 patients undergoing radical resection and 2 patients undergoing palliative resection. Forty-two patients underwent intraoperative combined resection of the adjacent organs. The recurrence and survival status of patients were followed up.Results:All the 89 patients underwent the operation successfully, with the median operative blood loss of 200 (10-2000) ml. There were 23 cases being diagnosed of well differentiated liposarcoma, 40 cases of dedifferentiated, 20 cases of myxoid/round, 5 cases of myxoid liposarcoma, and 1 cases of mixed type. Pathologically, there 42 cases with low grade histology and 47 cases with high grade histology. In this study, 89 patients were followed up for 3 to 108 months, and the median follow-up time was 28 months. The 5-year recurrence free survival rate, disease-free survival rate and overall survival rate of the patients were 16.7%, 16.1% and 52.6%, respectively. There were 57 patients presenting local recurrence, 1 patient of lung metastasis, and 1 patient of liver metastasis, and the median disease-free survival time was 24 months. There were 42 patients died of the disease, with a median survival time of 64 months. Univariate analysis showed that intraoperative blood loss( P<0.01), whether multiple cases( P<0.01), pathologic types( P<0.01), and histological grades ( P<0.01) were related to disease-free survival.The intraoperative blood loss( P<0.01), multiple cases( P<0.05), pathologic types ( P<0.05), and recurrence ( P<0.01)were related with overall survival. Gender, age, tumor size, tumor location, whether primary surgery, radical resection or combined resection of adjacent organ had no effect on the prognosis of patients ( P>0.05). Cox regression model multivariate analysis revealed that surgical bleeding ( RR=2.360, 95% CI 1.313-4.241, P=0.004), multiple tumor ( RR=1.899, 95% CI 1.068-3.375, P=0.029), and pathological type ( RR=4.976, 95% CI 1.622-15.264, P=0.005) were independent factors affecting disease-free survival. The recurrence was an independent factor affecting the overall survival of patients ( RR=31.495, 95% CI 1.062-933.684, P=0.046). Conclusions:Retroperitoneal liposarcoma is a rare disease with high recurrence rate. The intraoperative blood loss, whether multiplicity and pathological type are independent factors affecting the disease-free survival, and recurrence is independent factors affecting the overall survival.

The experiment was conducted by directed evolution strategy (error-prone PCR) to improve the activity of aflatoxin detoxifzyme with the high-throughput horse radish peroxidas and recessive brilliant green (HRP-RBG) screening system. We built up a mutant library to the order of 10(4). Two rounds of EP-PCR and HRP-RBG screening were used to obtain three optimum mutant strains A1773, A1476 and A2863. We found that mutant A1773 had upper temperature tolerance of 70 degrees C and that its enzyme activity was 6.5 times higher than that of the parent strain. Mutant strains A1476 worked well at pH 4.0 and its enzyme activity was 21 times higher than that of the parent strain. Mutant A2863 worked well at pH 4.0 and pH 7.5, and its enzyme activity was 12.6 times higher than that of the parent strain. With DNA sequencing we found that mutant A1773 revealed two amino acid substitutions, Glu127Lys and Gln613Arg. Mutant A1476 revealed four amino acid substitutions: Ser46Pro, Lys221Gln, Ile307Leu and Asn471lle. Mutant A2863 revealed four amino acid substitutions: Gly73Ser, Ile307Leu, Va1596Ala and Gln613Arg. The results provided a useful illustration for the deep understanding of the relationship between the function and structure of aflatoxin detoxifzyme.
Aflatoxin B1 ; antagonists & inhibitors ; chemistry ; Amino Acid Substitution ; Directed Molecular Evolution ; Enzyme Activation ; Enzyme Stability ; Multienzyme Complexes ; genetics ; metabolism ; Mutant Proteins ; genetics ; metabolism ; Point Mutation ; Polymerase Chain Reaction ; methods ; Protein Engineering

Objective To explore the DNA expression of human cytomegalovirus (HCMV) in glioma and the association between HCMV infection and prognosis of glioma patients.Methods Used for this study were 89 specimens of glioma which had been surgically ablated and pathologically confirmed from the patients between January 2007 and December 2016 at Department of Neurosurgery,The First Affiliated Hospital of Zhengzhou University,and Department of Neurosurgery,The First Hospital of China Medical University.Of them,32 belonged to WHO grade Ⅱ,31 to WHO grade Ⅲ and 26 to WHO grade Ⅳ.Ten specimens of normal brain tissue were excised as controls from the contemporary patients receiving resection for essential epilepsy.Nested PCR was used to analyze the DNA expression of HCMV in the glioma tissue and normal brain tissue,and in the peripheral blood from the glioma and control patients.Prognosis of the glioma patients was evaluated using the Kaplan-Meier survival analysis.Results The DNA expression of HCMV was positive in 46 of the 89 specimens of glioma,involving 14 cases of WHO grade Ⅱ,16 ones of WHO grade Ⅲ and 16 ones of WHO grade Ⅳ.The DNA expression of HCMV was negative in all the 10 specimens of normal brain tissue.There was a significant difference in the DNA expression of HCMV between the glioma tissue and normal brain tissue (P=0.002).The HCMV DNA was measured in the peripheral blood from 26 glioma patients,involving 10 cases of WHO grade Ⅱ,8 ones of WHO grade Ⅲ and 8 ones of WHO grade Ⅳ.No HCMV DNA was detected in the peripheral blood from the 10 control patients.There was a significant difference between the brain glioma and control groups in gene expression of HCMV in peripheral blood (P=0.048).There were no significant differences in the survival rate between the patients with positive or negative DNA expression of HCMV in the glioma tissue or in the peripheral blood from the glioma and control patients (x2=1.849,P=0.174;x2=0.082,2=0.774).Conclusion HCMV infection may play an active role in pathogenesis and development of glioma.

Objective : To investigate the changes in the sagittal diameter of the upper airway before and after the treatment of skeletal Class Ⅲ malocclusion in adults with microimplant anchorage and class Ⅲ intermaxillary elastics and to provide a reference for clinical treatment. Methods : A total of 35 adult patients with skeletal Class Ⅲ malocclusion were selected to be treated with the straight-wire technique. Microimplant group, 15 cases (group A): patients with severe skeletal Class Ⅲ malocclusion (vertical high angle) were treated with the straight-wire technique combined with microimplant anchorage; class Ⅲ intermaxillary elastics group, 20 cases (group B): Patients with mild or moderate skeletal Class Ⅲ malocclusion (vertical low angle and average angle) were treated with the straight-wire technique combined with class Ⅲ intermaxillary elastics, and cephalometric radiographs obtained before and after treatment in the upper airway in the two groups were measured and analyzed. Results : Changes in cranial and maxillofacial measurements after correction: in group A, (sella-nasion-supramental angle) the SNB angle decreased significantly (P < 0.05), and (subspinale-nasion-supramental angle) the ANB angle increased significantly (P < 0.05). In group B, the SNB angle decreased significantly (P < 0.05), while (subspinale-nasion-subspinale angle) the SNA angle、ANB angle and anterior skull base plane-mandibular plane angle (Sn-MP) angle increased significantly (P < 0.05). Changes in sagittal diameter of the upper airway measurements after corrections: In group A, the width of the glossopharyngeal segment of the upper airway (TB-TPPW) decreased significantly (P < 0.05). In group B, first segment width of the upper airway behind the hard palate (PNS-R) increased significantly (P < 0.05). After correction, the decreased SNB and increased ANB in group A was higher than that in group B, and the difference was statistically significant （P < 0.05）. The decreased of TB-TPPW in upper airway of group A was greater than that of group B, and the difference was statistically significant （P < 0.05）. Conclusions In the treatment of skeletal class Ⅲ malocclusion with microimplant anchorage, the sagittal diameter of the glossopharyngeal segment of the upper airway has a negative impact.