Objective To investigate the features of immune response of human immunodeficiency virus type-1 (HIV-1) antigen specific T lymphocytes in HIV-1 monoinfected or HIV 1/hepatitis C virus (HCV) coinfected individuals.Methods Twenty-six HIV-1 monoinfected and 23 HIV-1/HCV coinfected individuals were enrolled.Immunomagnetic microbeads were used to isolate T lymphocyte subpopulation CD4+ T cells and CD8+ T cells from human peripheral blood mononuclear cells (PBMC).Frequencies of interferon-γ (IFN-γ) secreting cells of CD4+,CD8+ T lymphocytes and PBMC stimulated by a peptide pool containing 12 overlapping peptides in HIV-1 P24 from 49 patients were assessed by enzyme-linked immunospot (ELISPOT) assay.HIV-1 RNA levels of these patients were also detected by real-time fluorescence quantitative polymerase chain reaction.The data were compared by one-way ANOVA and Mann-Whitney U test,and Spearman test was used for correlation analysis.Results Frequencies of HIV-1 antigen specific CD4+ T lymphocytes [median =25 spot-forming cells (SFC)/106 cells] were significantly lower than those of CD8+T lymphocytes (median=38SFC/106 cells,F=4.592,P=0.037) and PBMC (median=53 SFC/106 cells,F=5.436,P=0.025) in HIV-1 monoinfected group.Frequencies of HIV-1 antigen specific CD4+ T lymphocytes (median=5 SFC/106 cells,Z=-2.432,P=0.015),CD8+T lymphocytes (median=5 SFC/106 cells,Z=-1.996,P=0.046) and PBMC (median=10 SFC/106 cells,Z=-2.306,P=0.021) in HIV-1/HCV coinfected group were significantly lower than those in HIV-1 monoinfected group.Conclusions In HIV-1 infection,antigen specific immune response of CD4+ T cells can be activated,but weaker than that of CD8+ T cells.Co-infection with HCV might down-regulate the responses of HIV-1 antigen specific T lymphocytes in HIV-1 infected individuals.

Objective To investigate impacts of chronic hepatitis B virus (HBV) co-infection on immune responses and clinical manifestations in acute infection of dengue virus type 1.Methods The serum levels of interferon (IFN)a,IFNβ,IFNγ,interleukin (IL)-10 and tumor necrosis factor (TNF)α of 310 dengue serotype 1 (DENV1)-infected patients were assessed by enzyme linked immunosorbent assay (ELISA),among which 8％ (25/310) were chronic HBV co-infection.Meanwhile,serum samples from 41 healthy adults and 47 chronic HBV infected subjects were recruited as controls.Comparisons among groups were done by one factor analysis of variance and correlation analysis of results was done by spearman rank correlation test.Results The serum level of IFN-α [(95.1 ± 279.3) pg/mL] was significantly higher than IFN-β[(2.8 ± 16.2) pg/mL] during acute dengue infection,while IFN-α level [(86.5±358.1) pg/mL] reduced in patients with HBV co-infection.The secretion kinetics of IFNα,IFNγ (pro-inflammatory cytokine) and IL-10 (anti-inflammatory cytokine)were analyzed.The medians of IFNα,IFNβ and IL-10 level were elevated to peak on day 2,day 3-4and day 6 after fever onset,respectively.Additionally,IFNα levels in patients with only dengue infection were negatively correlated with the platelet counts and serum alanine aminotransferase levels (r=-0.2327,0.2122,both P＜0.01).Conclusions Chronic HBV co-infection alters human immune responses elicited by acute dengue viral infection.Moreover,IFNα secretion may be associated with hemorrhagic tendency,while protective against inflammatory damage of liver.

Objective To construct a prokaryotic expression vector pET-22b+with Middle East respiratory syndrome ( MERS) coronavirus nuclocapsid protein( NP) gene and to express and purify N protein.Methods N gene amplified by PCR was inserted into the prokaryotic expression vector pET-22b+.Recombinant plasmid was confirmed using DNA elec-trophoresis and sequencing.NP was expressed in E.coli BL21(DE3) by IPTG induced and purified by cation exchange chromatography using the AKTA purification system.Results The NP gene sequence was proved to be correct by sequen-cing and the protein was expressed in both soluble and insoluble forms in E.coli BL21 ( DE3 ) after IPTG induction.The purity and concentration of recombinant protein was improved obviously by cation exchange chromatography and enrich-ment.Conclusion Recombiant NP of high purity and concentration is purified and will facilitate NP functional research.

Objective To detect the impact of reactive oxygen species ( ROS) on mitochondrial morphology and distri-bution during mitosis.Methods A viral vector in which the fluorescence gene was specifically under the control of mito-chondrial promoter was constructed and confirmed through DNA sequencing and Western blotting.After transfecting HeLa s3 cell with packaged virus, the HeLa s3-COX4tp-EGFP cell line stably expressing the mitochondrial fluorescence signal was obtained.With immunofluorescent staining, the impact of ROS on the morphology and distribution of mitochondria dur-ing mitosis was inspected.Result The cell line constantly expressing mitochondrial fluorescence signals was successfully constructed.Meanwhile,it was found that H2 O2 treatment could significantly change the morphology and distribution of mi-tochondria during mitosis by confocal microscopy.Conclusion Our study demonstrates that ROS can affect the morphology and distribution of mitochondria during mitosis.This research help study the relationship between the mitochondrial function and the regulation of mitosis in the future.

Objective To evaluate the clinical value of ankle arthroscopy in diagnosis and treatment of Danis-Weber type B ankle fracture associated with injury to the distal tibiofibular syndesmosis.Methods A retrospective study was conducted of the 35 patients who had been treated at Department of Orthopaedics,Ruijin Hospital North for Danis-Weber type B ankle fracture from February 2014 to December 2016.They were 23 males and 12 females,with an average age of 43.1 years (from 18 to 65 years).Each of them underwent 4 examinations to detect whether injury to the distal tibiofibular syndesmosis was complicated or not:preoperative CT and MRI,C-ann roentgenography and ankle arthroscopy before and after internal fixation of the ankle.The diagnostic rates of the complicated injury by the 4 examinations were compared.The patients complicated with injury to the distal tibiofibular syndesmosis received surgical repair of the injury using TightRope in addition to internal fixation of the ankle,and injury to the deltoid ligament was repaired simultaneously using 3.5 mm anchor nails in case the injury was concomitant.The repair and stability of the distal tibiofibular syndesmosis were observed using ankle arthroscopy again.Results The Cotton and external rotation tests under C-arm roentgenography before surgery and after internal fixation of the ankle demonstrated that 13 cases were complicated with injury to the distal tibiofibular syndesmosis.Of the other 22 patients who had not been diagnosed with the injury by C-arm roentgenography,6,13 and 11 were diagnosed with the injury respectively by CT,MRI and ankle arthroscopy.The diagnostic rates of Danis-Weber type B ankle fracture associated with injury to the distal tibiofibular syndesmosis were 37.1％ (13/35),54.3％ (19/35),74.3％ (26/35) and 68.6％ (24/35) respectively by C-arm roentgenography,CT,MRI and ankle arthroscopy.In the sensitivity to the injury,MRI ＞ ankle arthroscopy ＞ CT ＞ C-arm roentgenography,with significant differences between them (P ＜ 0.05).The ankle arthroscopy confirmed the repair efficacy in the 24 patients complicated with injury to the distal tibiofibular syndesmosis and negative results of Cotton and external rotation tests in them.Additionally,ankle arthroscopy revealed 16 cases of injury to the deltoid ligament at the medial ankle.Conclusions Ankle arthroscopy can provide evidence for correct diagnosis and treatment of Type B ankle fracture complicated with injury to the distal tibiofibular syndesmosis,because it allows direct observation of the medial deltoid ligament of the ankle and the distal tibiofibular syndesmosis.It can be also used to assess the stability of the tibiofibular syndesmosis after repair of the injury.

Objective:Anterior cruciate ligament injury is the most common type of knee joint ligament injury.Anterior cruciate ligament reconstruction has a high failure rate,with bone tunnel abnormalities as the most significant factor in these failures.Digital orthopedic technology can effectively develop implementation plans for the revision,thus increasing the success rate.This study aims to develop a surgical plan for anterior cruciate ligament revision by employing multiplanar reconstruction(MPR)for measuring bone tunnel position and diameter,and simulating bone tunnel creation via 3D printing preoperatively. Methods:A total of 12 patients who underwent anterior cruciate ligament revision at the Third Xiangya Hospital of Central South University between 2014 and 2021 were retrospectively studied.The data included patient demographics,preoperative formulated knee joint 3D printing models,and preoperative knee CT scans.The study measured the bone tunnel's diameter and position to guide the establishment of revision bone tunnels during surgery,reassessed the postoperative bone tunnels,and evaluated knee joint functional scores[including International Knee Documentation Committee Knee Evaluation Form(IKDC)score,Lysholm score,and Tegner exercise level score]. Results:Preoperative measurements revealed suboptimal femoral tunnels positions in 4 patients and tibial tunnels positions in 2 patients.MPR and 3D printing technology were used to guide the establishment of a new bone canal during surgery,and postoperative measurements were satisfactory for all patients.Preoperative measurements demonstrated the interclass correlation coefficient for femoral tunnels and tibial tunnels diameters were 0.843(P<0.05)and 0.889(P<0.001),respectively.Meanwhile,the intraclass correlation coefficient were 0.811(P<0.05)and 0.784(P<0.05),respectively.The intraoperative diameter of femoral and tibial tunnels showed excellent correlation with postoperative CT measurements,with intraclass correlation coefficient values of 0.995(P<0.001)and 0.987(P<0.001),respectively.All bone tunnel positions were within the normal range.At the final follow-up,knee joint function scores in all 12 patients improved significantly compared to pre-surgery(P<0.001),and the reoperation rate was zero. Conclusion:MPR and 3D printing technology can accurately measure the parameters of reconstructed anterior cruciate ligament bone tunnels.Personalized revision plans for patients with reconstruction failure enhances the success rate of revision surgery and improves patient prognosis.

Objective: To investigate clinical value of ankle arthroscopy in diagnosis of type Danis-Weber B ankle fracture with the distal tibiofibular syndesmosis injury. Methods: From August 2014 to January 2018, a total of 52 cases of type Danis-Weber B ankle fractures with an average age of 35.37±11.19 years, including 31 males and 21 females were treated. Each patient underwent preoperative assessment, according to the patient's clinical manifestations and imaging examination before operation. The Cotton test and the external rotation test were conducted in the C arm X-ray to evaluate whether there is the distal tibiofibular syndesmosis injury. All fractures were treated with open reduction and internal fixation. Repair was conducted with 3.5 mm wire anchor and nail for triangular ligament injury. The Cotton test and external rotation test were performed under ankle arthroscopy. The injury of the lower tibiofibular syndesmosis was observed and evaluated by cotton test and external rotation test in patients who were found to be associated with lower tibiofibular joint injury. After internal fixation of fracture and repair of tibiofibular syndesmosis injury with TightRope, the repair effects of tibiofibular syndesmosis injury was observed under direct vision of ankle arthroscopy. The VAS score, AOFAS score and Baird-Jackson score was used as an index to observe the pain and ankle function before and after operation for 12 months follow-up. Results: Preoperative X-ray showed 19 cases of lower tibiofibular joint injury. Preoperative CT showed 28 cases of lower tibiofibular joint injury. Preoperative MRI showed 39 cases of lower tibiofibular joint injury. A total of 36 cases of joint injury of lower tibiofibular syndesmosis were confirmed under arthroscopy of ankle. After repair of tibiofibular syndesmosis injury with TightRope, complete reduction and stability of inferior tibiofibular syndesmosis were confirmed under ankle arthroscopy. At 12 months after operation, the VAS score decreased from 8.13±1.32 points preoperation to 0.75±0.73 points after operation. The AOFAS and Baird-Jackson scores increased from 26.59±3.35 points, 16.44±3.63 points preoperation to 94.36±3.03 points, 90.29±6.45 points after operation, respectively. There was significant difference before and after operation (P<0.05). The postoperative VAS score, AOFAS score and Baird-Jackson score of 16 patients without lower tibiofibular ligament injury were confirmed in the ankle arthroscopy group, namely 0.58±0.67 points, 95.27±4.67 points, 91.98±9.23 points. Conclusion Ankle arthroscopy can be used to observe the injury of tibiofibular syndesmosis under direct vision. The positive rate of diagnosis of inferior tibiofibular syndesmosis injury was 69.2% (36/52). It can provide reliable evidence for correct diagnosis and treatment of type Danis-Weber B ankle fracture with lower tibiofibular joint injury with significant meaning in selecting operative methods and in evaluating the effect of lower tibiofibular ligament repair.

【Objective】 To investigate the application value of 450 nm blue laser vaporization in the treatment of benign prostatic hyperplasia (BPH) in district hospitals, so as to provide reference for the selection of BPH treatment methods in district-level hospitals. 【Methods】 The clinical data of 39 BPH patients who received surgical treatment in Chang’an Hospital during Jan.and Sep.2023 were retrospectively analyzed.The patients were divided into two groups, including 19 in blue laser group treated with 450 nm blue laser vaporization and 20 in the TURP group treated with transurethral resection of the prostate (TURP).The operation time, hemoglobin decline, postoperative bladder irrigation time, catheter indwelling time and adverse events were compared between the two groups. 【Results】 All operations were successfully completed.In the blue laser group, the operation time [(45.11±22.08) min vs.(81.25±29.37) min], postoperative bladder irrigation time [(34.05±9.41) h vs.(47.60±9.05) h] and the decrease of hemoglobin [(7.79± 5.86) g/L vs.(16.40±10.23) g/L] were significantly lower than those in the TURP group (P<0.05).There was 1 case of urinary retention in the blue laser group, and there was no significant difference in indwelling catheter time between the two groups (P>0.05). 【Conclusion】 The 450 nm blue laser vaporization of prostate has advantages of less blood loss, shorter operation time and shorter postoperative irrigation time.Therefore, it should be considered as a preferable option for BPH in primary hospitals.