Objective To investigate the effect of acute exposure to cadmium combined with bacitracin on the endoplasmic reticulum stress (ERS) in testes and ovaries of rats and its regulation by nuclear factor erythroid-2-related factor 2 (Nrf2). Methods According to the 4×2 factorial design model, 48 specific pathogen free adult SD rats were divided into four groups: the control group and the low-, medium- and high- dose cadmium chloride exposure groups. Each group was further divided into with- or without bacitracin combined subgroup. There were six rats in each subgroup with 3 males and 3 females. The low-, medium- and high- dose groups were intraperitoneally injected with 5, 10, 20 mg/kg body weight of cadmium chloride solution, respectively. The control group was intraperitoneally injected with the same amount of 0.9% sodium chloride solution. Among them, rats in the bacitracin combined subgroup were given a one-time intraperitoneal injection of bacitracin at a dose of 20 mg/kg body weight two hours before cadmium chloride exposure. After 48 hours, the rats were sacrificed. The mRNA expression of glucose regulated protein78 kD (Grp78), protein kinase R-like endoplasmic reticulum kinase (Perk), Nrf2 in testes and ovaries of rats was determined using quantitative real-time polymerase chain reaction. The protein expression of GRP78, PERK, NRF2 was determined using Western blotting. Results The mRNA expression of Grp78, Perk, Nrf2 and the protein expression of GRP78, PERK, NRF2 in testes and ovaries of rats in the no bacitracin combined subgroups of the three dose groups showed different degrees of up-regulated changes compared with the no bacitracin combined subgroup of the control group (all P<0.05). Among them, the expression of the three kinds of mRNAs and proteins in the testes and ovaries of rats in the no bacitracin combined subgroups of the high-dose group was up-regulated (all P<0.05), and most of them were higher than those in the no bacitracin combined subgroups of the low- and medium-dose groups (all P<0.05). The expression of most of the three kinds of mRNAs and proteins in testes of rats showed different degrees of down-regulated changes (all P<0.05), but the expression of the three kinds of mRNAs and proteins showed different degrees of up-regulated changes in ovaries (all P<0.05) in the bacitracin combined subgroups of the three doses groups than that in the bacitracin combined subgroups of the control group, and especially in the bacitracin combined subgroups of the high-dose subgroup. The expression of the three kinds of mRNAs and proteins in testes and ovaries of rats in the bacitracin combined subgroups of the three doses groups showed different degrees of changes (all P<0.05) compared with the no bacitracin combined subgroup in the same group, and the expression in the bacitracin combined subgroups of the medium- and high-dose groups showed mainly down-regulated changes (all P<0.05). Conclusion Acute exposure to cadmium can induce different degrees of ERS, activate PERK/NRF2 signaling pathway, and improve the toxicity to testis and ovary. Bacitracin can inhibit cadmium-induced ERS, thereby inhibiting the activation of PERK/NRF2 signaling pathway, and enhancing the synergistic effect of cadmium on testis and ovary toxicity. The higher the exposure dose of cadmium, the more obvious the inhibitory effect.

Objective: To explore neural correlates for the recognition of facial expression videos using event-related functional magnetic resonance imaging.Methods:Thirteen right-handed healthy Chinese women underwent blood oxygen level dependent (BOLD) fMRI voluntarily while recognizing happy, sad, neutral emotional faces and fixation cross videos.Results:In comparison with fixation cross, recognition of neutral faces activated the left middle frontal gyrus, the bilateral precentral gyrus, the right amygdala, the left parahippocampal gyrus, the right postcentral gyrus and the left thalamus. In comparison of neutral faces recognition, the recognition of happy faces elicited increased activation in the right medial frontal gyrus, the right superior frontal gyrus, the middle frontal gyrus, the right anterior cingulated gyrus, the left subcallosal gyrus, the right superior occipital gyrus, the left middle occipital gyrus and the right superior temporal gyrus; while the recognition of sad faces activated the left medial frontal gyrus, the right middle frontal gyrus, the left inferior temporal gyrus and the left superior temporal gyrus.Conclusion:This study indicates that the recognition of facial expression is mediated by a distributed cortical network, and the medial frontal gyrus may be involved in discriminating different affective information, while the superior temporal gyrus may play an important role in processing the dynamic characters of face.

Objective To investigete the effect of low-concentration ozone (O3) on voltage-gated calcium channels in fetal rat spinal cord neurons in vitro. Methods The spinal cord neurons were cultured in vitro and identified by immunohistochemistry staining. Then the cells were seeded into the culture dish and randomly divided into 3 groups: control group (group C, n = 5), O3 15 μg/ml (group O3-15, n = 4) and O3 20 μg/ml group (group O3-20, n = 8). The cells were exposed to O3 15 and 20 μg/ml in group O3-15 and O3-20 respectively, while the cells were exposed to air in group C. After 20 min incubation, the electrophysiological activity of calcium channels in neurons was recorded using whole-cell patch-clamp technique. Results Peak calcium current density was significantly increased in group O3-15 and O3-20 compared with group C ( P ＜ 0.05). Half-activation voltage was significantly lower in group O3-15 and O3-20 than in group C ( P ＜ 0.05). Peak calcium current density was significantly higher in group O3 -20 than in group O3-15 ( P ＜ 0.05) . There was no significant difference in half-inactivation voltage among the three groups ( P ＞ 0.05). Conclusion Low concentration of O3 can make the activation of the calcium ion channel easy in fetal rat spinal cord neurons in vitro.

Objective To construct a geometric skull model by using three-dimensional reconstruction, computer tomography scanning and rapid prototyping technology and evaluate its significance in treatment of complex oral and maxillofacial deformities. Methods A cranial and a goldenhar syndrome patient with complex oral and maxillofacial deformities bone received continuous volumetric scanning of skull and the data acquisition was done by an electron computed tomography, by which reconstruction was performed and the obtained images saved as STL files. Then the data were input into rapid prototyping machine to make three-dimensional geometric model. Direct measurement, designation and surgery simulation could be done on this three-dimensional model. Then the mandibular mirror physical model was manufactured using rapid prototyping according to the normal side. Results A computer-aided model according to CT data could represent the three-dimensional anatomic structures and their relationships precisely. The replica exhibited dimensional errors ranging 0.02 mm to 0.53 mm. Which provided strong basis for accurate understanding of disease status and reasonable surgical plans and helped improve the curative effect of surgery. Conclusion Rapid prototyping can help surgeons in many ways for therapy of the complex oral and maxillofacial deformities.

Objective To evaluate the effect of injection of ozone (O3) through lateral recess for the treatment of radiculitis caused by lumbar disc herniation on the blood glucose in patients with diabetes mellitus.Methods Ninety-six patients with radiculitis caused by lumbar disc herniation involving 48 patients with diabetes mellitus and 48 patients without diabetes mellitus were enrolled in the study.The diabetic or non-diabetic patients were randomly divided into 2 groups ( n =24 each):non-diabetic patients-O3 group (group N-O3 ),non-diabetic patients-ghcocorticoid group (group N-GC),diabetic patients-O3 group (group D-O3 ),and diabetic patients-glucocorticoid group (group D-GC).In N-O3 and D-O3 groups,30 mg/L O3 10 ml was injected via the lateral recess.In N-GC and D-GC groups,compound betamethasone injection 3.5 mg was injected via the lateral recess.The blood glucose level was measured before treatment (T1),1 and 4 h after treatment (T3,4),and 1,3 and 7 days after treatment (T5-7).VAS score was recorded at T1,immediately after treatment (T2),and at T5-7.The patients' quality of life was measured using the Medical Outcomes Study 36-Item Short-Form Health Survey (MOS SF-36 Health Survey) questionnaire at T1,7.The therapeutic effect was evaluated at T7.The side effects were recorded.Results Compared with the baseline value at T1,VAS scores were significantly decreased after treatment,while the MOS SF-36 Health Survey questionnaire scores for physical functioning,bodily pain,role emotional,and mental health in all groups,for general health in group N-GC,and for vitality in groups D-O3 and N-O3 were significantly increased at T7,and the blood glucose level was significantly increased at T4 in groups N-GC and D-O3,and at T3-7 in group D-GC ( P ＜ 0.05).There was no significant change in the blood glucose level before and after treatment in group N-O3.The blood glucose level was significantly higher in group N-GC than in group N-O3,and in group D-GC than in group D-O3 ( P ＜ 0.05).There were no significant differences in the excellent and good rates and all the MOS SF-36 Health Survey questionnaire scores among all groups.No side effects were found in the study.Conclusion Injection of ozone through lateral recess for treatment of radiculitis caused by lumbar disc herniation exerts no influence on the blood glucose level in patients with diabetes mellitus.

BACKGROUND: Recombinant adeno-associated virus 2(rAAV-2) has attracted considerable attention due to its nonpathogenic nature in contrast to other viral vectors such as adenoviral and retroviral vectors in gene therapy attempts.OBJECTIVE: To explore rAAV-2 transduction to bone marrow mesenchymalstem cell(BMSC) in vitro and evaluate the possibility of using rAAV-2 as a vector for gene therapy of acute myelogenous leukemia(AML).DESIGN: An open experiment with cells as the observational subjects.SETTING: Department of Hematology, Nanfang Hospital, Southern Medical University.MATERIALS: The experiment was conducted in the Department of Hematology, Nanfang Hospital, Southern Medical University from February to July 2004. We used passages 3 to 5 BMSCs derived from six de novo AML patients and four healthy volunteers in this study.METHODS: BMSC was isolated from 6 to 10 mL of bone marrow aspirates obtained from the iliac crests of the patients who had been diagnosed as having de novo AML and from those of healthy volunteers. The acquired BMSC was infected by rAAV-2 which contained enhanced green fluorescent protein (rAAV-2-eGFP) at different multiplicity of infection(MOI) (MOI = 1 × 102,1 × 103, 1 × 104, 1 × 105, 1 × 106, 1 × 107) . Then we observed through phase contrast fluorescent microscope and flow cytometer to evaluate green fluorescent protein(GFP) expression 10 to 14 days after transduction. GFP expression was observed as the rAAV-2-eGFP transduced BMSC cultured in vitro. We also observed the in vitro gene expression profile of GFP in rAAV-2-eGFP transduced BMSC which was selected by neomycin ( G418). First, we confirmed GFP expression in BMSC through phase contrast fluorescent microscope, then on flow cytometer to detect the percentage of GFP expression.MAIN OUTCOME MEASURES: The efficiency of rAAV-2-eGFP transduction to BMSC. GFP expression was observed through phase contrast fluorescent microscope and flow cytometer at different time points after transduction.rAAV-2-eGFP to BMSC derived from normal volunteers and AML patients had no significant differences. GFP began to express 10 to 14 days after transduction, and the transduction efficiency ranged from 0. 3% to 1.4%. By changing infection condition, we could not make a higher transduction efficiency( P ＞ 0.05) . One round infection of BMSC by rAAV-2-eGFP at a MOI of 1 × 105 was ( 1. 030 ± 0. 034) %, 3 rounds of infection of BMSC by rAAV-2-eGFP at a MOI of 1 × 105 was (1. 140 ±0. 036)%, and coinfected by LipofectAMINE was (1. 380 ± 0. 054)%. However, 293 cell line which was the package cell of rAAV-2 could be efficiently transduced by AML patients transduced by rAAV-2-eGFP at MOI = 1 × 105: The percentage of GFP expression cell gradually decreased from 1.14% at day 12 after transduction to 0. 6% as cell passaged from 2 to 3, and maintained at a level of 0. 5% to 0. 6% later on till 61 days after transduction. After selected by neomycin(G418) 1 month later, rAAV-2-eGFP transduced BMSCs could maintain a long-term GFP expression at a level of 6.0% in vitro without significant decay within 100 days of observation period after transduction.CONCLUSION: The advantages of rAAV-2 mediated gene transduction lie in safety, no immune response to the host, and long-term expression maintained by the target gene. rAAV-2 and BMSC can be used for in vitro gene therapy, and as a systemic gene delivery system, it might be an alternative for systemic gene therapy in the future.

Objective: This study was designed to investigate the variations of endotoxin,glutamine and TNF-? concentration in septic rats. Methods: A total of 90 adult male SD rats were divided as follows: control group(normal),experimental group(CLP).They were killed on hour 6,12,24,48,72 after surgery.Plasma,liver,small intestine and skeletal muscle were collected to measure the concentrations of glutamine in plasma and tissues,the levels of endotoxin and TNF-? in plasma. Results: In septic rats,plasma concentrations of endotoxin and TNF-? increased on hour 6,significantly increased on hour 12,attained the peak on hour 24,and decreased evidently on hour 48 and 72.Glutamine concentration in plasma elevated on hour 6 and 12,elevated significantly on hour 24 and 48,and decreased on hour 72.Glutamine concentration in liver increased on hour 6～12,increased significantly on hour 24,and decreased significantly on hour 48 and 72.Glutamine concentration in small intestine and skeletal muscle decreased on hour 6 and 12,and decreased significantly after 24 hour. Conclusion: During the early stage of sepsis,the plasma levels of endotoxin and TNF-? are increased significantly,the glutamine concentration in plasma and liver is also increased significantly,however,it was decreased evidently in skeletal muscle and small intestine.

Objective:To investigate the relationship between the expression of serum exsomal miRNA-155-5p (miR-155-5p) and prognosis in patients with esophageal squamous cell carcinoma (ESCC).Methods:A total of 336 samples from ESCC patients in Fujian Provincial Cancer Hospital from October 2014 to December 2015 were collected. The relative expression levels of serum exsomal miR-155-5p were detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). Cut-off value of the expression levels of serum exsomal miR-155-5p was determined by using X-tile software. Based on the optimal cut-off value, patients were divided into miR-155-5p low expression group and miR-155-5p high expression group. The survival curve was drawn by using Kaplan-Meier method and Cox proportional hazards regression model was used to make survival analysis.Results:The cut-off value of serum exsomal miR-155-5p expression level was 2.340. According to the cut-off value, patients were divided into miR-155-5p low expression group (<2.340) of 51 cases and miR-155-5p high expression group (≥2.340) of 285 cases. There were no statistical differences in age ( χ2 = 0.020, P = 0.887), gender ( χ2 = 0.283, P = 0.595), tumor location ( χ2 = 0.063, P = 0.977), differentiation grade ( P = 0.474), clinical staging ( χ2 = 3.996, P = 0.136) and surgery treatment ( χ2 = 0.941, P = 0.332) of patients in both groups. ESCC patients in serum exsomal miR-155-5p high expression had a higher risk of death compared with patients in miR-155-5p low expression group ( HR = 1.763, 95% CI 1.049-2.961, P = 0.032). Conclusion:The high expression level of serum exsomal miR-155-5p is associated with poor prognosis in ESCC patients and it could be used as a prognostic new marker in ESCC patients.

Objective To investigate the effects of ozone (O3) on Wnt/β-catenin signaling pathway in the articular cartilage of rats with osteoarthritis (OA).Methods Eighteen male SPF Wistar rats,aged 3 months,weighing 200-250 g,were randomly divided into 3 groups (n =6 each) using a random number table:control group (group C),group OA,and O3 group (group O).OA was induced by injection of monosodium iodoacetate 3 mg (50 μl) into the right knee joint cavity.On 7th day after the model was established successfully,25 μg/ml O3 1 ml were injected into the knee joint cavity,once a week for 3 consecutive weeks in group O.Behavioral changes were observed after establishment of the model.At 1 day before establishment of the model,and 1,4,7,14,21 and 28 days after establishment of the model,the mechanical paw withdrawal threshold (MWT) was measured.At 28 days after establishment of the model,the total knee joint was removed and stained with haematoxylin and eosin for examination of the pathological changes of the cartilage (under light microscope) and for determination of the expression of β-catenin and matrix metalloproteinase-13 (MMP-13) in the cartilage (by immunohistochemistry).Results The signs of OA such as hind-limb motor dysfunction,knee joint swelling,or decreased joint motion,and signs of hyperalgesia such as lickings were observed after establishment of the model in rats.Compared with group C,the MWT was significantly decreased at each time point after establishment of the model,and the ex pression of β-catenin and MMP-13 in the cartilage was significantly up-regulated in the other two groups(P＜0.05).Compared with group OA,the MWT was significantly increased at 7-28 days after establishment of the model,and the expression of β-catenin and MMP-13 in the cartilage was significantly down-regulated in group O (P＜0.05).The pathological changes of the cartilage were significantly reduced in group O as compared with group OA.Conclusion The mechanism by which O3 mitigates OA is probably related to inhibition of Wnt/β-catenin signaling pathway activation in the articular cartilage in rats.

Objective To analyze CMV DNA stability of 30 EDTA plasma samples in the order of magnitude between 300 and 100 000 copies/mL over a 21 day period. Methods Thirty plasma samples were grouped into three categories according to the CMV DNA loads , including low CMV DNA contents , intermediate CMV DNA loads and high CMV DNA loads. Ten milliliters of whole blood was freshly collected from each patient. Plasma samples without hemolysis were divided into 1-ml aliquots. One aliquot was processed immediately (Day 0) for baseline PCR assays. The remaining aliquots were then processed after one , two, three, seven, 14 or 21 day of storage at 4℃. Results There was no significant difference between the mean of the difference time point in viral loads following storage at 4 ℃ by paired-samples t test, including Day 1 compared to Day 0 (t = 1.654, P =0.109), Day 2 compared to Day 0 (t = 1.487, P = 0.148), Day 3 compared to Day 0 (t = 1.609, P = 0.118), Day 7 compared to Day 0 (t=0.831, P=0.413), Day 14 compared to Day 0 (t=1.721, P=0.096), and Day 21 compared to Day 0 (t=0.244, P=0.810). Conclusion The concentration of CMV DNA in all samples stored at 4 ℃ for 21 days did not differ significantly from the baseline viral load ,and it was not observed the trend in continued degradation in different time point (Day 1, 2, 3, 7 and 14).