Objective:To investigate the eye movement characteristics of non-emotional pictures and emotion-al pictures in college students with different self-rating levels of depression.Methods:The Baker Depression Ques-tionnaire(BDI-2)was used to select 20 college students in high score group(BDI-2 score≥ 18 points)and 20 col-lege students in low score group(BDI-2 score ≤2 points).In experiment 1 the different finding tasks were used to investigate the difference of eye movement features between the two groups when viewing non-emotional pic-tures.In experiment 2 the expression recognition tasks were used to investigate the difference of the eye movement features between the two groups when viewing emotional pictures.Results:The results of experiment 1 showed that when completing the judgment task,the fixation times of each interest area of non-emotional images was smaller in the high score group than in the low score group(P＜0.01).The results of experiment 2 showed that the reaction time was longer to sad faces than to happy faces in the high score group(P＜0.05),and the correct rate to sad faces was higher in the high score group than in the low score group(P＜0.05).For the eye interest area,the first fixation arrival time was earlier in the high score group than in the low score group(P＜0.001),and the first fixa-tion time and total fixation time were shorter in the high score group than in the low score group(Ps＜0.01).Con-clusion:College students with high self-rating level of depression show a decrease of interest when viewing non-e-motional pictures,and show an advantage in processing negative emotional information and avoiding eye gaze when viewing emotional pictures.

Objective:To investigate the role of dendritic cells (DC) in Chlamydia muridarum ( Cm) respiratory infection and their effect on adaptive immune response. Methods:C57BL/6 mice were exposed to 1×10 3 inclusion-forming units (IFU) of Cm through inhalation to establish the mouse model of Cm respiratory infection. The proportion of CD11c + MHCⅡ + DC and the expression of costimulatory molecules (CD40, CD80 and CD86) in spleen tissues were detected by flow cytometry on 0, 3 and 7 d after infection. The expression of IL-12p40, IL-10 and IL-6 at mRNA level in spleen tissues was detected by qPCR. Mouse splenic DC isolated on 7 d after Cm infection were sorted by magnetic beads and then transferred to recipient mice. Th1 response in the recipient mice was measured using intracellular cytokine staining 14 d after infection. Results:Cm respiratory infection induced massive infiltration of DC and promoted the expression of costimulatory molecules on splenic DC. The expression of IL-12 and IL-10 at mRNA level in splenic DC reached the peak on 3 d after infection. Transferring the splenic DC of Cm-infected mice into the recipient mice could alleviate the disease condition in the recipient mice after Cm infection with reduced Cm inclusion-forming units in lung tissues and significantly increased proportion of Th1 cells in lung and spleen tissues. Conclusions:Cm respiratory infection could induce the maturation and activation of DC, which promoted Th1 immune response. DC played an important role in Cm infection.

Objective:To investigate the efficiency and safety of traditional growing rod in the treatment of early onset dystrophic scoliosis secondary to type 1 neurofibromatosis (NF1-DS) with intraspinal rib head in children.Methods:From September 2006 to May 2020, this study recruited 20 children with intraspinal rib head with early onset NF1-DS who had received traditional growing rods. There were 13 boys and 7 girls and the age of the initial operation was 7.0±1.6 years (range, 4.1-9.8 years). There were 7 cases of simple left chest bend, 9 cases of simple right chest bend, and 4 cases of double chest bend; 13 patients had varying degrees of kyphosis deformity. Two children had neurological symptoms before surgery, American Spinal Injury Association Impairment Scale (AIS) were grade D. The proportion of the intraspinal rib head (IRP), the Cobb angle of the main chest bend, apical vertebra rotation (AVR), apical vertebral translation (AVT), trunk shift (TS) and sagittal TK, lumbar lordosis (LL), sagittal balance and T 1-S 1 height were measured before and after first time internal fixation and at last follow-up, and the complications were also evaluated. Results:All 20 patients were followed up and the average follow-up time was 41.6±23.8 months (range, 24-99 months). A total of 85 operations was conducted including 63 protrude operations. After operation, the IRP was significantly lower than that before operation (preoperative 33.1%±17.5% vs. postoperative 22.2%±11.3%, P<0.001) and no significant correction loss was found at last follow-up 23.7%±12.4% ( P>0.05). The mean Cobb angle decreased from 75.9°±26.7° preoperatively to 45.0°±18.5° postoperatively ( P<0.001) and there was still significant improvement at the last follow-up (41.0°±17.2°) compared with postoperatively ( P<0.05). The AVR was significantly reduced after surgery compared with preoperatively (33.0°±10.1° vs. 39.3°±13.3°, P<0.001), and the last follow-up (40.1°±11.4°) was significantly improved compared with postoperative ( P=0.005). The T 1-S 1 height increased from 259.8±70.7 mm preoperatively to 296.9±78.4 mm postoperatively ( P=0.001), and at the last follow-up 296.9±78.4 mm was still significantly higher than after operation ( P<0.001), with an average annual increase of 12.4±3.2 mm. Significant correction of AVT, TK, LL and sagittal balance were noted after initial surgery ( P<0.05), and no significant correction loss was found at last follow-up ( P>0.05). There were 10 complications in 7 cases. There were 5 complications of pedicle screw loosening, 1 complication of bolt droping, 2 complications of broken rod, 1 complication of distal junctional kyphosis and 1 complication of adding-on phenomenon. 2 cases with nerve injury were recover after operation (AIS grading E). None of the children had new neurological complications during growth rod insertion and multiple stretching during follow-up. Conclusion:For children with early onset NF1-DS with intraspinal rib head, if the preoperative AIS grade is D or E, traditional growing rod technique is relatively safe and effective and can make the intraspinal rib head remove from the spinal canal partly.

Objective:To explore the clinical efficacy of posterior short-segment internal fixation for the treatment of brucella spondylitis (BS).Methods:The medical records of 34 patients with BS admitted from January 2014 to June 2019 were retrospectively analyzed. There were 22 males and 12 females; the age was 52.3±10.6 years (range 35-72 years). On the basis of standardized use of antibacterial drugs, the lumbar spine posterior short-segment internal fixation was used. Twenty-nine cases underwent simple internal fixation, and posterolateral bone graft fusion, while 5 cases underwent primary debridement, autologous bone grafting and interbody fusion. Monitor erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and test tube agglutination test (SAT) were used to assess inflammation control. Imaging examinations of patients before operation, 1 month after operation, 3 months after operation, 6 months after operation, 1 year after operation to the last follow-up were analyzed to evaluate the condition of intervertebral fusion. The clinical efficacy evaluation was based on the pain visual analog scale (VAS), Japanese Orthopaedic Association (JOA) score, modified MacNab grading, and American Spinal Injury Association (ASIA) grading, as well as surgery-related complications.Results:The operation time of 34 patients was 104.64±16.72 min (range 65-145 min), the average hospital stay was 16.49±7.41 days (range 7-38 d), and the average postoperative follow-up time was 20.2 months (range 12-34 months). At the last follow-up, the ESR and CRP fell to the normal range, and the SAT was negative. At 3 months postoperatively, 11 cases (32.35%) reached Bridwell fusion criteria of grade II, 23 cases (67.65%) of grade III; 3 cases (8.82%) of grade I fusion at 6 months after surgery, 31 cases reached grade II fusion (91.18%); all reached grade I fusion at the last follow-up. After the operation, the symptoms of the waist or lower extremities were significantly relieved. The VAS score was 6.3±1.4 before the operation, 4.1±1.2 at 1 month after the operation, 2.7±1.4 at 3 months after the operation, 1.6±1.0 at 6 months after the operation, and 1.2±0.8 at the last follow-up. The JOA score before surgery was 13.8±2.4, 1 month after surgery 17.6±2.6, 3 months after surgery 21.7±3.1, 6 months after operation 4.9±2.7, and at the last follow-up 25.7±1.8. Compared with the preoperative time nodes of the above indicators, the differences were statistically significant. At the last follow-up, of the 12 patients (2 cases of grade C, 10 cases of grade D) with preoperative neurological dysfunction, 2 cases recovered from grade C to grade D, and 10 cases recovered from grade D to E; the excellent and good rate of modified MacNab grading reached 97.06% (33/34). No extradural hematoma, nerve damage, cerebrospinal fluid leakage and other surgical complications occurred. Only 1 case had wound infection complication, and the prognosis was good after active treatment. There were no recurrences during the follow-up period.Conclusion:On the basis of standardized antimicrobial treatment, posterior lumbar short-segment internal fixation is a safe and effective method for the treatment of BS, and good clinical effects can be obtained.

Hepatitis C virus (HCV) infects more than 200 million people globally and its increasing incidence is placing a heavy economic burden on resource-poor countries.Effective HCV diagnosis is of guiding significance for treatment selection and monitoring.Techniques for detection of HCV infection include serological and nucleic acid detection.Serological assay is simple and easy to operate for initial HCV diagnosis.However,the window phase is long,so the active phase of the virus or previous infection cannot be determined.Nucleic acid detection has high sensitivity and wide linear range,which is the direct evidence to determine virus infection.The traditional thermal cycle-based nucleic acid amplification technology has been widely used in clinical laboratories.The development of isothermal nucleic acid amplification technology overcomes the limitations of traditional amplification technology,and it can be used as a tool for Point-of-care Testing (POCT),which has great potential and particularly suitable to be used in low-resource areas with high HCV prevalence,such as Africa and Southeast Asia.In this paper,we outline the current advance and development trends in the HCV molecular diagnosis.

Objective: To compare the detection rate of herpes virus and enterovirus (EV) in paired cerebrospinal fluid and serum samples of patients with viral encephalitis. Methods: A total of 109 paired cerebrospinal fluid and serum specimens were collected from patients who were clinically diagnosed with suspected viral meningitis in Children′s Hospital of Hunan from December 2017 to February 2018. One-step nested real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) and real-time fluorescence quantitative PCR were used to detect enterovirus and herpes virus respectively and the detection rates of different virus and sample types were analyzed. SPSS 17.0 was used for statistical analysis of the test result . Results: Among the 109 pairs of specimens, the positive rates of human herpes virus type 6 (HHV6), herpes simplex virus-1 (HSV1), Epstein-Barr virus (EBV), cytomegalovirus (CMV) and enterovirus group A type 71(EV-A71) in serum were 7.34%, 4.59%, 7.34%, 9.17% and 10.09%, respectively, and in cerebrospinal fluid were 5.50%, 2.75%, 0, 5.50%, and 6.42%, respectively. The result showed that there were statistically significant differences between the two types of specimens for herpes virus and enterovirus (P<0.05). In cerebrospinal fluid and serum samples, the longest time for EV-A71 positive detection was 2 and 7 days after onset, respectively; the longest time for CMV positive detection was 3 and 26 days after onset, respectively; the longest time for HHV6 positive detection was 7 and 8 days after onset, respectively; the longest time for HSV1 positive detection was both 12 days after the onset; in serum samples, the longest time for EBV positive detection was10 days after onset, but in cerebrospinal fluid, no EBV was detected within 10 days of onset. Conclusions EV-A71 is the most prevalent pathogen causing viral encephalitis in hunan, the overall positive rate of virus in serum samples was higher than that in cerebrospinal fluid samples. Virus stays longer in serum than in cerebrospinal fluid. It is suggested that the time is of great significance for the pathogen detection of children with viral encephalitis, the specimen type can be selected reasonably according to the time of onset.

Objective: To establish a real-time fluorescence recombinase acid amplification (RAA) method for the detection of adenovirus type 3(HAdV-3)without extraction nucleic acid. Methods: According to the conserved sequence of adenovirus type 3 gene, a pair of primers and a probe were designed, and a real-time fluorescence RAA without extracting nucleic acid was established and optimizing the condition of DNA-free extraction. The sensitivity of the method was analyzed by a series of dilution and the specificity of the method was evaluated by detecting the original samples of other respiratory viruses. The clinical samples of HAdV-3 were detected and compared with the traditional real-time fluorescence quantitative PCR method for nucleic acid extraction. Results: The sensitivity of the real-time fluorescence RAA method was as high as that of qPCR in the detection of 10 series diluted HAdV-3 strains. The highest corresponding CT value of qPCR was 36.87. The sensitivity of the real-time fluorescence RAA method was similar to that of the real-time fluorescence quantitative PCR method . There was no cross-reaction to other common types of respiratory viruses. The two method were used to detect 56 clinical samples at the same time, and the result were completely consistent. Conclusions We provide the first report of the real-time fluorescent RAA assays for the detection of HAdV-3 without extracting nucleic acid and it has high sensitivity and specificity. Is suitable for rapid detection of HAdV-3 in clinical laboratories and on-site unite.

Objective To investigate the effect of acidic tumor microenvironment on invasion and migration and its mechanism in glioma cells. Methods (1) The pH value of the medium was adjusted by acid-base titration. Human glioma cells U87 and U251 were cultured in the acid group and the normal group with pH values of 6.4 and 7.4, respectively; and 3 d after cultivation, the expressions of hypoxia-inducible factor-2α (HIF-2α) and CD44 were detected by Western blotting; Transwell assay was used to examine the invasion and migration of U87 and U251 cells; immunofluorescence was employed to examine the CD44 expression. (2) The U87 and U251 cells were divided into small interfering RNA (siRNA) -nonsense sequence group and siRNA-CD44-1 group, and the siRNA nonsense sequences and siRNA-CD44-1 interfering fragments were transfected by lipofectin-3000, respectively; three d after transfection, the migration and invasion abilities of cells from the two groups were detected by Transwell assay. (3) U87 and U251 cells were divided into acid group (cultured with a pH value of 6.4), blank control group, siRNA nonsense sequence group, siRNA-CD44-1 group, and siRNA-CD44-2 group; and cells from the later four groups were cultured with a pH value of 7.4; after culture for 4 d, the siRNA-nonsense sequence group, siRNA-CD44-1 group and siRNA-CD44-2 group were transfected with siRNA-nonsense sequences, siRNA-cd44-1 interfering fragments and siRNA-CD44-2 interfering fragments, respectively; three d after transfection, the expressions of CD44, N-Ca, Vimentin, and matrix metalloproteinase (MMP)-2 proteins in these 5 groups were detected by Western blotting. Results (1) As compared with the normal group, the expression levels of HIF-2α and CD44 in U87 and U251 cells of the acid group were significantly increased; both Transwell and invasion experiments showed that the number of transmembrane cells in the acid group was significantly larger than that in the normal group (P<0.05); immunofluorescence staining showed that the CD44 expression in acid group was significantly higher than that in normal group (P<0.05). (2) Both Transwell and invasion experiments showed that the number of transmembrane cells in the siRNA-CD44-1 group was significantly smaller than that in the siRNA nonsense sequence group (P<0.05). (3) Western blotting showed that the expression levels of CD44, N-Ca, Vimentin and MMP-2 in U87 and U251 cells of the blank control group, siRNA nonsense sequence group, siRNA-CD44-1 group, and siRNA-CD44-2 group were obviously decreased as compared with those in the acid group; the expression levels of CD44, N-Ca, Vimentin and MMP-2 in U87 and U251 cells of the siRNA-CD44-1 group and siRNA-CD44-2 group were obviously lower than those in the siRNA nonsense sequence group. Conclusion Acidic tumor microenvironment enhances the capabilities of invasion and migration of glioma cells through increasing CD44 expression.

Although oxymatrine (OMT) has been shown to directly inhibit the replication of hepatitis B virus (HBV), limited research has been done with this drug. In the present study, the antiviral effect of OMT was investigated in an immunocompetent mouse model of chronic HBV infection. The infection was achieved by tail vein injection of a large volume of DNA solution. OMT (2.2, 6.7 and 20 mg/kg) was administered by daily intraperitoneal injection for 6 weeks. The efficacy of OMT was evaluated by the levels of HBV DNA, hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and hepatitis B core antigen (HBcAg). The immunoregulatory activity of OMT was evaluated by serum ELISA and flow cytometry. Results shows that OMT at 20 mg/kg inhibited HBV replication, and it was more efficient than entecavir (ETV) in the elimination of serum HBsAg and intrahepatic HBcAg. In addition, OMT accelerated the production of interferon-(IFN-) in a dose-dependent manner in CD4T cells. Our findings demonstrate the beneficial effects of OMT on the enhancement of immunological function and in the control of HBV antigens. The findings suggest this drug to be a good antiviral therapeutic candidate for the treatment of HBV infection.

Objective To study calcium chelator BAPTA-AM antagonize the cellular oxidative stress induced by hydrogen peroxide ( H2 O2 ) and to explore the effect of calcium ion on the cell degeneration mediated by NLRP3.Methods The SHSY5Y cell model of oxidative stress was made by hydrogen perox-ide,then the cell model was treated with calcium ion carrier A23187 or BAPTA-AM,a higher efficiency cal-cium chelating agent.The cells were divided into 4 groups:H2O2 treatment group,H2O2+A23187 group, H2 O2+A23187 +BAPTA-AM group and control group.NLRP3 protein was detected by Western blot,and Caspase-1 and IL-1βwere detected by ELISA.Results NLRP3 expression was significantly increased in cells treated by hydrogen peroxide(P<0.05) .The NLRP3 protein continued to increase, and the expression of Caspase-1((57.1±19.2)pmol/L) and IL-1β((484.2±49.5)pg/ml) protein was also increased signifi-cantly in cells treated by A23187,and the difference had statistically significant for caspase-1 or IL-1βin H2O2+A23187 group compared with those in control group(Caspase-1:(26.8±12.9)pmol/L,IL-1β:(326.9 ±52.1) pg/ml) (P<0.05, P<0.01) .NLRP3,Caspase-1 and IL-1βwere all significantly reduced after adding a high-er efficiency calcium chelating agent BAPTA-AM.Conclusion Calcium overload is likely to enhance the oxidative stress induced by hydrogen peroxide and engender neurodegeneration mediated by NLRP3 inflammasome.