[ ABSTRACT] AIM:To study the effects of nuclear factor kappa B ( NF-κB) on human hepcidin expression in fer-ric ammonium citrate ( FAC)-induced HH4 hepatocytes.METHODS:Non-transformed HH4 cells were exposed to FAC at concentrations of 0.1, 1, 5 and 10 mmol/L for 48 h.The expression of iron regulatory gene hepcidin was determined by semi-quantitative RT-PCR.The effects of NF-κB on hepcidin transcriptional activity were detected using chromatin immuno-precipitation (ChIP), electrophoretic mobility shift assay (EMSA) and dual-luciferase reporter assay system, combined with the inhibition experiments of intracellular NF-κB activity.RESULTS: FAC at concentrations of 5 mmol/L and 10 mmol/L significantly enhanced the expression of hepcidin.The results of ChIP and EMSA showed the binding of NF-κB to the upstream of hepcidin promoter.Treatment with NF-κB inhibitor BAY 11-7082 attenuated hepcidin expression.The lucif-erase activity in the cells transfected with recombinant luciferase reporter plasmid was obviously higher than that in control group.CONCLUSION:NF-κB is the transcription factor that contributes to hepcidin expression in iron overload-induced HH4 cells.

Objective To observe the effect of Jingui Shenqi Wan (JSW) on the content of calcium channel protein CaV1.3 of guinea pigs with ototoxic deafness induced by gentamicin (GM). Methods Forty healthy guinea pigs were randomly divided into normal group, model group and high-, middle-, and low-dose JSW treatment groups. The model group was given intramuscular injection of GM ( 120 mg/kg) per day for 10 continuous days. The high-, middle-, and low-dose JSW treatment groups were given intramuscular injection of GM (120 mg/kg) and intragastric administration of JSW in the dosage of 20.3, 13.5, 6.08 g·kg-1·d-1 respectively per day for 10 days. Immuno histochemistry was used to detect the mean optical density value of CaV1.3 expression in cochlear hair cells. Results There were significant differences of the optical density value of CaV1.3 in cochlear hair cells between the model group and normal group (P<0.01), and between the model group and high-dose JSW treatment group (P<0.05). Conclusion JSW has certain effect on preventing and treating guinea pig with ototoxic deafness induced by GM, thus to protect the hearing function.

Objective To present the management of increasing the closure stress of urethra for treatment of stress urinary incontinence in women. Methods In 12 patients with stress incontinence the posterior urethra and the anterior wall of the bladder was incised.Then the wall of posterior urethra,bladder neck and bladder trigone were trimmed and sutured to form a tube whereby to elongate and tighten the posterior urethra referring to Campbell-Young's way and according to Laplace's law. Results The mean period of post-operative follow-up was 8.8 years.Eleven patients could completely control their urinating without residual urine after the operation.The short-term and long-term outcomes were the same in these 11 patients.For the remaining one patient little urine was spilt when the abdominal pressure was increased with exertion. Conclusions Elongating and tightening the posterior urethra is simple,effective and safe for treatment of stress urinary incontinence in women.

The proteomics definition, investigation method and its a pplication in cancer research were simply introduced. Proteomic research is to r eveal the function of genes from an integrated, kinetic and quantitative view at the global protein level, which is an important component of post-genome proje ct. Cancer is a kind of complex disease involved by multi-genes. Proteomic rese arch will be helpful to discover the mechanism of cancer development, to find sp ecial malignant tumor markers and targets of drug treatment.

Objective:To investigate the effect of miRNA-1-3p (miR-1-3p) on expression of myocyte enhancer factor 2A (MEF2A) and the biological function of osteosarcoma cells.Methods:The tumor tissues and adjacent normal tissues of 20 patients with osteosarcoma who were clinically diagnosed in Shanxi Provincial Cancer Hospital from January 2019 to January 2020 were collected, and the expression of miR-1-3p in the samples was detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The expression of miR-1-3p in osteosarcoma cell lines U2-OS, SAOS-2, MG63, SW1353 and human normal osteoblast cell line hFOB1.19 was detected by qRT-PCR, then the cell line with the lowest expression of miR-1-3p was selected for follow-up experiments. An overexpression miR-1-3p vector was constructed (miR-1-3p mimcs). The miR-1-3p overexpression group was transfected with miR-1-3p mimcs, and the control group was transfected with empty vector (miR-1-3p nc). CCK-8 method was used to detect the proliferation activity of cells; flow cytometry was used to detect the changes of cell apoptosis and cell cycle. miRwalk database was used to predict the miR-1-3p target gene, and the target gene was verified by dual-luciferase reporter gene assay; Western blot was used to detect the expression of MEF2A protein in cells of each group.Results:Compared with adjacent tissues, the expression of miR-1-3p in osteosarcoma tissues was down-regulated (0.31±0.14 vs. 0.62±0.21), and the difference was statistically significant ( t = 5.31, P<0.01). The expression of miR-1-3p in U2-OS cells was the lowest; compared with the control group, the proliferation activity of U2-OS cells was inhibited in miR-1-3p overexpression group (48 h absorbance value 0.56±0.01 vs. 0.77±0.03, t = 2.77, P<0.01; 72 h absorbance value 0.87±0.02 vs. 1.40±0.03, t = 2.93, P<0.01); G 1/S cell cycle arrest increased [G 1 phase (38.24±0.55)% vs. (32.11±0.80)%, t = 9.27, P = 0.01; S phase (61.24±0.90)% vs. (67.78±0.83)%, t = 7.52, P = 0.02]; early apoptotic rate increased [(11.20±0.12)% vs. (1.50±0.12)%, t = 2.91, P<0.05], miRwalk database predicted that the miR-1-3p target gene was MEF2A. The result of dual-luciferase reporter gene assay showed that miR-1-3p bound to MEF2A 3'UTR, and the luciferase activity of U2-OS cells in miR-1-3p overexpression group was lower than that in the control group (renilla luciferase/firefly luciferase activity ratio 0.53±0.06 vs. 1.00±0.04, t = 4.04, P < 0.05). Western blot showed that the expression of MEF2A protein in U2-OS cells of miR-1-3p overexpression group was lower than that of the control group (protein relative expression 0.41±0.14 vs. 0.77±0.12, t = 3.93, P < 0.05). Conclusions:The low expression of miR-1-3p may be associated with the proliferation, apoptosis and cycle changes of osteosarcoma cells. miR-1-3p can negatively regulate the expression of MEF2A protein and regulate the occurrence and development of osteosarcoma.

Objective To investigate the relationship of the serum bilirubin level with acute ischemic stroke (AIS), and Chinese isch-emic stroke subclassification (CISS), stroke severity and short-term outcome of AIS patients. Methods 616 patients with AIS as well as 664 patients without stroke matched with gender and age were compared and analyzed with the non-conditional Logistic regression. The AIS pa-tients were divided based on the CISS, and their bilirubin levels were compared. The AIS patients were divided into high bilirubin group and normal bilirubin group, their scores of the National Institute of Health Stroke Scale (NIHSS) as admission and discharge, and neural func-tion improvement rate were compared. Results The levels of total bilirubin (TBIL), direct bilirubin (DBIL) and indirect bilirubin (IBIL) were higher in the AIS group than in the control group (P<0.01), and the TBIL was the independent risk factors of AIS (OR=1.026, 95%CI 1.012-1.041, P<0.001). There was no significant difference of the levels of TBIL, DBIL and IBIL among the patients of CISS (P>0.05). The NIHSS score was higher in the high bilirubin group than in the normal bilirubin group as admission (P<0.05), but it was not significant as discharge (P>0.05), nor the rate of neural function improvement (P>0.05). Conclusion The serum bilirubin level elevated and correlated with the severity in the AIS patients, which might be the risk of pathogenesis and AIS. The bilirubin was not various with the CISS, and might be less involved in the short-term outcome of AIS.

Objective To investigate the risk factors of recurrent ischemic stroke, and evaluate the severity and short-term outcome. Methods From March, 2014 to March, 2015, 238 patients with recurrent ischemic stroke and 378 patients with initial ischemic stroke matched with gender and age were enrolled. The clinical data of two groups were compared and the non-conditional Logistic regression model was made to analysis the risk factors of recurrent ischemic stroke. The National Institute of Health Stroke Scale (NIHSS) score as ad-mission and discharge, and the neural function improvement rate of two groups were compared. Results The incidences of leukoarailsis (χ2=8.666), hypertension (χ2=8.189), smoking (χ2=6.973) and alcohol consumption (χ2=4.722) were higher in the recurrent group than in the pri-mary group (P<0.05). Logistic regression analysis showed that leukoarailsis (OR=1.690, 95%CI:1.198~2.384, P=0.003), hypertension (OR=1.715, 95%CI:1.135~2.592, P=0.010) and smoking (OR=1.896, 95%CI:1.233~2.915, P=0.004) were the independent risk factors of recur-rent ischemic stroke. The NIHSS scores as admission and discharge were significantly higher (t=-3.645, t=-4.675, P<0.001), and the neural function improvement rate was lower (t=2.643, P<0.01) in the recurrent group than in the primary group. Conclusion Hypertension, leuko-arailsis and smoking are independent risk factors for recurrent ischemic stroke. Recurrent ischemic stroke is more serious than initial isch-emic stroke, and the short-term neural function recovers more slowly.

Neural cell adhesion molecule (NCAM) is a glycoprotein expressing on the surface of neurons, glial cells, bone cells and natural killer cells. NCAM plays an important role in the process of cell - cell adhesion and cell migration, and is also a model protein to study polysialic acid. In this paper, NCAM gene from mouse mammary gland cells (NMuMG) was cloned into eukaryotic expression vectors pcDNA3.1(+) and transfected into mutant Chinese hamster ovary cells ldlD-14. The stable transfection over-expressing NCAM was obtained through the G418 selection and confirmed by Western blotting. Due to unique characters of ldlD-14 cells, carbohydrate chain of NCAM molecule can be easily manipulated with or without adding galactose in the serum free medium, and this modification can provide the basis for further studies on the effect of glycosylation on NCAM molecular function.
Animals ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Female ; Galactose ; Glycosylation ; Mammary Glands, Animal ; cytology ; Mice ; Neural Cell Adhesion Molecules ; biosynthesis ; Polysaccharides ; chemistry ; Sialic Acids ; chemistry ; Transfection

Objective To investigate the clinical efficacy,operative essentials and indications of hybrid spinal fusion surgery for cervical spondylotic myelopathy.Methods From August 2008 to December 2011,thirty-eight patients with cervical spondylotic myelopathy underwent hybrid spinal fusion surgery in our hospital.There were 27 males and 11 females,aged from 33 to 70 years (average,51 years).A total of 86 segments were treated (fusion 48 vs.non-fusion 38).Twenty-eight patients underwent a two-level surgery,and ten patients received a three-level surgery.The Japanese Orthopaedic Association (JOA) score and Visual analogue scale (VAS) were used to evaluate pre-and post-operative neurological function and pain,respectively.The pre-and post-operative range of motion of the cervical spine was measured according to Xrays.Moreover,the surgical complications were recorded and analyzed.Results Thirty-seven patients were followed up for 15 to 55 months (average,29.1 months).The improvement of neurological function was obtained in 36 patients.The JOA score was improved from preoperative 10.5±1.57 to 14.3±1.97 at final follow-up,with an improvement rate of 58.46％,and the results were excellent in 16 cases,fair in 20 cases and poor in 1 case.The VAS was improved from preoperative 7.3±1.04 to 3.2±1.41 at final follow-up.The Cobb angle changed from preoperative 25°±3.21°to 20°±2.56°at final follow-up.After operation,the neurological function was not restored in 1 case; hoarseness and bucking occurred in 2 cases; sore throat occurred in 22 cases; anterior displacement of prosthesis (PCM) occurred in 3 cases.Other patients had no complications,such as displacement,loosening and heterotopic ossification.Conclusion In hybrid spinal fusion surgery,the lesions segments are decompressed fully,the severely degenerative segments are fused,and the motion of the non-fusion segments is reserved.As a result,not only the stability of the cervical spine is achieved,but also an obvious improvement of symptoms and a satisfactory short-term efficacy can be obtained.Therefore,this method is an alternative procedure for cervical spondylosis myelopathy.

Purpose To investigate TRAP1 expression in esophageal cancer and its relationship with clinicopathological features and prognosis.Methods Expression levels of TRAP1 in 60 pairs of cancer and adjacent normal tissues were detected by immunohistochemistry,and the relevance between TRAP1 and clinicopathological features was evaluated.Kaplan-Meier and Cox proportional regression analyses were performed to determine the association of TRAP 1 expression and survival of the patients.Results The positive expression rate of TRAP1 was 55.0％,and the amount of relative protein transcript level was 2.7 ± 1.1 in the cancer tissue.The positive expression rate of TRAP1 was 11.7％,and the relative expression protein was 0.5 ± 0.4 in the adjacent normal tissue.The expression level of TRAP1 in cancerous tissue was significantly higher than that of tissue adjacent to carcinoma.There was no statistically significant difference between the expression levels of TRAP1 with sex,age,location,and degree of differentiation (P ＞ 0.05).There was statistically significant relationship between recurrence,metastasis and TNM stages with the expression of TRAP1 levels (P ＜ 0.05).The average survival time of TRAP1-negative patients was 69.0 months (95％ CI:60.2-77.9) while the TRAP1-positive patients was 34.2 months (95％ CI:24.4-44.1).High levels of TRAP1 were correlated with decreased survival of postoperative esophageal cancer patients (P ＜ 0.05).Conclusion TRAP1 is overexpressed in esophageal cancer and associated with the progression and prognosis of esophageal cancer.