In order to develop a positive quality control products for the detection of porcine repro-ductive and respiratory syndrome virus(PRRSV)nucleic acid by real-time fluorescent quantitative PCR(RT-qPCR),the positive quality control products of PRRSV-1 and PRRSV-2 M genes were prepared using armored RNA technology of MS2 phage.PRRSV-1 and PRRSV-2 M genes were amplified,purified and recovered,and ligated into pET28b vector containing MS2 mature enzyme protein gene and capsid protein.After transformed into BL21(DE3),the gene products were in-duced by IPTG and purified by PEG6000 precipitation method to prepare the armored RNA virus-like particles(AR-PRRSV)containing PRRSV M gene.Following the performance evaluation,as the positive quality control products of PRRSV-1 and PRRSV-2 M genes,AR-PRRSV1M and AR-PRRSV2M were calculated using YY/T 1652-2019 standard.Results showed that it had a good u-niformity,stable storage for the armored virus-like particles at-20,4,25 ℃ for 60 d,and 37 ℃ for 30 d.The prepared armored virus-like particles AR-PRRSV1M and AR-PRRSV2M were deter-mined by digital quantitative PCR(ddPCR)after preliminary quantification by RT-qPCR.The 104 copies/μL of AR-PRRSV1M and AR-PRRSV2M ddPCR fixation was(1.33+0.50)× 104 cop-ies/μL.The above results indicates that the AR-PRRSVM can be used as the quality control of the whole detection process(nucleic acid extraction,reverse transcription and RT-qPCR).

Objective:To investigate the application of artificial intelligence based on the neural network radiation field in repair of soft tissue defects at lower limbs.Methods:A retrospective analysis was performed of the 23 patients who had been admitted to Department of Orthopedic Surgery, Renmin Hospital of Wuhan University from June 2020 to May 2022 for soft tissue defects at lower limbs. There were 14 males and 9 females, aged (38.6±6.7) years. Causes for soft tissue defects: traffic injury in 9 cases, benign or malignant primary soft tissue tumor in 6 cases, mechanical injury in 4 cases, crush injury in 2 cases, and chronic ulcer in 2 cases. Defect locations: the thigh in 3 cases, the lower leg in 7 cases, and the ankle and distal foot in 13 cases. The areas of soft tissue defect ranged from 6.0 cm×3.8 cm to 14.7 cm×12.8 cm. The defects were repaired and reconstructed by transplantation of an anterolateral femoral free flap in 7 cases and a pedicled flap in 16 cases with the assistance of artificial intelligence based on the neural network radiation field, a cutting-edge artificial intelligence algorithm that can quickly construct and process three-dimensional model images through volume rendering under the radiation field. The flap survival rate, aesthetic satisfaction before and after treatment, time for skin flap harvesting and transplantation, functional recovery of lower limbs and incidence of complications were recorded.Results:All the 23 patients were followed up for 32(28, 36) weeks. All the flaps were harvested smoothly and survived. The time for flap harvesting and transplantation was 65.8(50.0, 76.0) min. The aesthetic satisfaction scored (2.3±0.7) points before treatment and (8.4±1.6) points 4 weeks after treatment, showing a statistically significant difference ( P<0.05). The skin flaps healed well with no complications such as hematoma or infection in all but one patient who suffered from superficial necrosis at the distal skin flap due to venous crisis but healed with a scar. On average, the functional recovery of lower limbs scored 23.7(22.0, 25.0) points at 12 weeks after operation according to the Enneking evaluation system, and the functional recovery of lower limbs was 79% (23.7/30.0). Conclusion:Application of artificial intelligence based on the neural network radiation field can achieve ideal results in repair of soft tissue defects at lower limbs, due to its advantages of rapid and accurate surgical procedures, limited damage to the donor site, and a short learning curve.

BACKGROUND: Programmed cell death 1 or programmed cell death ligand 1 inhibitor (PD-1/PD-L1 inhibitor) and docetaxel, as the standard second-line treatments of advanced non-small cell lung cancer (NSCLC) patients, have limited effects. There are few studies on whether docetaxel combined with PD-1/PD-L1 inhibitor can increase the efficacy and make patients better benefit. The aim of this study is to evaluate the efficacy and safety of docetaxel combined with PD-1/PD-L1 inhibitor for the second-line treatment of stage IV NSCLC patients. METHODS: Stage IV NSCLC patients (n=118) who received treatment at Shandong Cancer Hospital between October 1, 2018, and December 31, 2020, were retrospectively analyzed. They were divided into observation group (n=69) and control group (n=49) according to different treatment plan. Observation group was given docetaxel plus PD-1/PD-L1 inhibitor, while control group was given PD-1/PD-L1 inhibitor. The clinical curative effect and the incidence of adverse reactions of grade 3 and above were compared between the two groups. RESULTS: The disease control rate (DCR) was higher in the observation group (89.9%) than that in the control group (73.5%) (P=0.019), and the objective response rate (ORR) showed no significant difference between observation group (24.6%) and control group (16.3%) (P=0.276). Till June 22, 2021, the 1-year PFS rate showed no difference between observation group (16.5%) and control group (7.7%) (P=0.205). During the treatment period, the adverse reactions of the two groups were mostly grade 1 to 2, and could be tolerated. The incidence of bone marrow suppression in observation group was higher than that in the control group (P<0.05), and the remaining adverse reactions were not statistically different from control group. Cox regression analysis showed that performance status (PS) (P=0.020) and age (P=0.049) were independent prognostic factors for the effect of docetaxel combined with PD-1/PD-L1 inhibitor. CONCLUSIONS The second-line treatment with docetaxel plus PD-1/PD-L1 inhibitor in patients with stage IV NSCLC can improve the DCR and prolong the PFS, and the adverse reactions are tolerable.

Objective:To compare the effects of conservative oxygen therapy and conventional oxygen therapy on the efficacy and prognosis of mechanical ventilation support in patients with severe pneumonia.Methods:A prospective randomized controlled study were conducted, 110 patients with severe pneumonia requiring mechanical ventilation in intensive care unit (ICU) of Hefei First People's Hospital from May 2019 to May 2021 were selected and divided into conventional oxygen therapy group (51 cases) and conservative oxygen therapy group (55 cases) according to random number table method. Patients in both groups were treated with invasive to non-invasive sequential mechanical ventilation strategy, and the target of oxygen therapy in the conventional oxygen therapy group was set as the arterial partial pressure of oxygen (PaO 2) > 150 mmHg (1 mmHg = 0.133 kPa) or pulse oxygen saturation (SpO 2) > 0.96. In the conservative oxygen therapy group, PaO 2 was set at 70-100 mmHg or SpO 2 at 0.90-0.92. The changes of blood gas analysis, mechanical ventilation time, ICU stay time, ICU mortality, new organ dysfunction and infection more than 48 hours after admission were observed in two groups before and after treatment. Kaplan-Meier method was used to analyze the difference in cumulative survival rate between the two groups. Results:After treatment, pH value, PaO 2 and oxygenation index (PaO 2/FiO 2) in both groups were significantly higher than before treatment, and arterial partial pressure of carbon dioxide (PaCO 2) was significantly lower than before treatment. pH value in the conservative oxygen therapy group was significantly higher than that in the conventional oxygen therapy group (7.4±0.1 vs. 7.3±0.3). PaO 2, PaCO 2 and PaO 2/FiO 2 were significantly lower than those in conventional oxygen therapy group [PaO 2 (mmHg): 68.9±4.7 vs. 75.2±6.0, PaCO 2 (mmHg): 42.1±5.6 vs. 50.5±7.5, PaO 2/FiO 2 (mmHg): 329±126 vs. 365±108, all P < 0.05]. The mechanical ventilation time in the conservative oxygen therapy group was significantly longer than that in the conventional oxygen therapy group (days: 19.7±3.5 vs. 13.9±4.5, P < 0.05), but there was no significant difference in ICU hospitalization time between the conservative oxygen therapy group and the conventional oxygen therapy group (days: 26.5±5.0 vs. 25.5±4.6, P > 0.05). Compared with the conventional oxygen therapy group, the ICU mortality, the incidence of nosocomial bloodstream infection, and the incidence of liver insufficiency, shock, and ICU acquired weakness (ICUAW) were decreased significantly in the conservative oxygen therapy group [ICU mortality: 27.3% (15/55) vs. 45.1% (23/51), liver insufficiency: 1.8% (1/55) vs. 7.8% (4/51), shock: 3.6% (2/55) vs. 9.8% (5/51), ICUAW: 5.5% (3/55) vs. 11.8% (6/51), the incidence of new bloodstream infection: 5.5% (3/55) vs. 11.8% (6/51), all P < 0.05]. Kaplan-Meier analysis showed that the cumulative survival rate of the conservative oxygen therapy group was significantly higher than that of the conventional oxygen therapy group (72.7% vs. 54.9%; Log-Rank test: χ 2 = 4.244, P = 0.039). Conclusion:Conservative oxygen therapy can reduce ICU mortality, the incidence of shock, liver insufficiency, ICUAW, and bloodstream infection in patients with severe pneumonia, but prolong mechanical ventilation time.

Objective:To investigate the effect of mannose on the radiosensitivity of six human non-small cell lung cancer cell lines and its possible mechanism.Methods:The expression of mannose phosphate isomerase in six lung cancer cell lines were detected by Western blot. The inhibitory effect of mannose on the proliferation of lung cancer cell lines were observed by MTT assay. When irradiated with 0, 2, 4, 6, 8 and 10 Gy, the effect of mannose on the radiosensitivity of six lung cancer cell lines was detected by plate clone formation assay, respectively; and the apoptosis rates of normal control, mannose, irradiation and combined groups were detected by flow cytometry.Results:The expression levels of mannose phosphate isomerase were different among six lung cancer cell lines. Among them, A549 cells had the highest expression level and H460 cells showed the lowest expression level. When aD ministrated with 11.1 mmol/L mannose, the same inhibitory effect was observed on both A549 and H460 cell lines. Moreover, the inhibitory effect on H460 cell line was significantly increased with the increase of mannose concentration. In addition, aD ministration of 11.1 mmol/L mannose could significantly increase the radiosensitivity and apoptosis rate of H460 cell line. However, it exerted limited effect upon the radiosensitivity and apoptosis rate of A549 cell line. Conclusion:In six lung cancer cell lines with high expression of mannose phosphate isomerase, the aD ministration of mannose can enhance the radiosensitivity of partial tumors cells.

OBJECTIVE： To investigate the improvement effects of external application of water extract from Picrasma quassioides on allergic contact dermatitis （ACD） model mice， and to provide reference for its further development and utilization. METHODS： A total of 48 mice were randomly divided into blank control group （normal saline）， model group （normal saline）， hydrocortisone group （positive control， about 0.15 g/ear） and water extract from P. quassioides low-dose， medium-dose and high-dose groups （0.15， 0.3， 0.6 g/ear， calculated by extract）， with 8 mice in each group. The mice were given medicine on ear at 8 a.m. and 4 p.m. for 7 d continuously. Except for blank control group， other groups were given acetone-olive oil solution containing 0.5％ 2，4-dinitrofluorobenzene （DNFB） on the abdomen of mice to induce ACD model. After 7 d medication， modeling group was given acetone-olive oil of 0.25％ DNFB on left ear to induce dermatitis. 24 h later， the pathological changes of left ear tissue were observed by naked eye and skin lesion was scored. The thickness difference and weight difference of left and right ears in mice were measured. The serum contents of IL-6 and IgE were determined. The pathomorphology changes of ear tissue were observed by microscope after HE staining. RESULTS： Compared with blank control group， the skin lesion score of left ear was increased significantly in model group （P＜0.01）； the thickness difference and weight difference of left and right ears， serum contents of IgE and IL-6 were increased significantly （P＜0.01）. Pathological changes were observed in left tissue of mice by microscope， such as lymphocytic infiltration， intercellular edema， erythrocyte exudation. Compared with model group， skin lesion score of left ear in mice was decreased significantly in hydrocortisone group and water extract from P. quassioides high-dose group （P＜0.05 or P＜0.01）； the thickness difference and weight difference of left and right ears in mice were decreased significantly in all administration groups （P＜0.01）. The serum contents of IgE and IL-6 in mice were decreased significantly in hydrocortisone group， water extract from P. quassioides medium-dose and high-dose groups （P＜0.01）； the inflammatory lesions of left ear in mice were alleviated to varying degrees in different administration groups. CONCLUSIONS： External application of water extract from P. quassioides has a good improvement effect on ACD model mice.

Tumor necrosis factor-α (TNF-α) is a key molecule of the inflammatory response, and plays a significant role in regulating cell survival and death.Many studies demonstrate that TNF-α may be implicated in the pathogenesis of various inflammatory conditions.Inhibiting the activity of TNF-α can dramatically improve the manifestations of many diseases.This review introduced the structural and functional characters of TNF-α and its receptors, and then summarized the regulating mechanisms TNF-α on cell survival、apoptosis and necroptosis.Besides, combining with the newly achievements of clinical and fundamental researches, this review analyzed the connections between TNF-α and common disorders in ophthalmology, such as the inflammation of retina and cornea, choroidal neovascularization, and summarized the clinical application and efficacy of infliximab, TNF-α monoclonal antibody.This review may provide references for the treatment of inflammatory diseases in ophthalmology and scientific researches in the future.

Objective To evaluate the effect of antiplatelet drugs and ischemic-events-free duration by thromboelastography (TEG) in patients undergoing peripheral endovascular treatment.Methods From Mar 2015 to Oct 2016,38 patients undergoing initial successful peripheral stenting and then coming back for recurrent ischemic events.TEG was used to evaluate the platelet inhibition rate induced by arachidonic acid (AA％) approach,the platelet inhibition rate induced by adenosine diphosphate (ADP％) approach,and the ADP-induced platelet-fibrin clot strength (MAADP).Clinical feature differences between patients with MAADP ＞ 47 mm and MAADP ≤ 47 mm were compared.Results AA％ less than 50％ was found in 9 patients(23.9％),and ADP％ less than 30％ in 14 patients(36.8％).5 patients met the two conditions.Patients with MAADp ＞ 47 mm had a significantly lower ADP％ (x2 =10.755,P ＜ 0.001),compared to patients with MAADP ≤47 mm.Univariate survival analysis showed ADP％ ＜ 30％ and MAADP ＞47 mm were risk factors for getting shorter ischemic-events-free duration,compared to patients with ADP％ ≥ 30 (P ＜ 0.001),with MAADP ≤47 mm (x2 =4.408,P =0.036).Conclusions TEG may detect those patients who has a low response to antiplatelet drugs,and some TEG parameters may have a ischemic predictive value.

Objective To study the association between serum testosterone level and insulin resistance (IR) in elderly male hypertensive patients.Methods One hundred and seven elderly male hypertensive patients were divided into IR group (n=44) and IR-free group (n=63).Their total TST level was measured,their FAI and TSI were assayed.The patients were further divided into TST≥14 nmol/L group (n=72) and TST＜14 nmol/L group (n=35).The association between serum testosterone level and IR in elderly male hypertensive patients was analyzed.Results The serum TST level,FAI and TSI were significantly lower in IR group than in IR-free group (12.02±2.66 nmol/L vs 15.98±3.98 nmol/L;20.16％±2.75％ vs 28.53％±4.74％;2.26±0.49 nmol/U vs 3.21±0.55 nmol/U,27.67％±5.49％ vs 25.98％±4.98％;2.95±0.39 nmol/U vs 2.78±0.64 nmol/U,P＜0.05).The FAI and TSI were significantly higher in hypertention grade 1 patients than in hypertension grade 3 patients (P＜0.05).The IR was significantly lower while the serum TST level,FAI and TSI were significantly higher in TST≥14 nmol/L group than in TST＜14 nmol/L group (33.3％ vs 57.1％,P=0.019;18.43±3.41 nmol/L vs 12.15±2.23 nmol/L,P=0.002;32.49％±5.67％ vs 24.57％±6.94％,P=0.036;3.53±0.87 nmol/U vs 2.55±0.62 nmol/U,P=0.016).TST,FAI and TSI were negatively associated with HOMA-IR (r=-0.406,r=-0.469,r=-0.429,P=0.000).Conclusion Low serum TST level is a risk factor for IRin elderly male hypertensive patients.

Objective To establish the apoptosis model in mouse cone cell line 661W cells and to investigate the viability of 661W cells in the conditions of different levels of autophagy.Methods Different concentrations of anti-Fas antibody were added to establish the apoptosis model of 661W cells,the expression of caspase-3 was detected by Western blot and the appropriate concentration of anti-Fas antibody was screened.Different concentrations of the autophagy inhibitor 3-methyladenine (3-MA) or autophagy inducer rapamycin were added to inhibit or induce autophagy,the expression of microtubule-associated protein 1 light chain 3 (LC-3) Ⅱ/LC-3 Ⅰ were detected by Western blot and the appropriate concentrations were also screened.The cultured cells were divided into 6 groups:control group,simple 3-MA group,simple rapamycin group,model group,model + 3-MA group and model + rapamycin group.Western blot was adopted to detect the expression of caspase-3,caspase-8,autophagy related genes 5 (Atg-5) and LC-3 Ⅱ/LC-3 Ⅰ at 0 hour,3,6,12,24 and 48 hours after induction.Flowcytometer was adopted to detect the apoptosis rate of 661W cells.Results The apoptosis model of 661W cells was successfully established,and the appropriate concentration of anti-Fas antibody was 2.0 μg/ml.After stimulated by the anti-Fas antibody,the expression of caspase-3 and caspase-8 of 661W cells increased from the time point of 6 hours,peaked at 12 hours,and sustained to 48 hours.However,the expression of Atg-5 and LC-3 Ⅱ/LC-3 Ⅰ raised from the time point of 3 hours,peaked at 24 hours,and decreased to the basic level at 48 hours.In addition,the appropriate concentrations of 3-MA and rapamycin were 20 nmol/L and 2.0 nmol/L,respectively.There was no statistical difference among the control group,simple 3-MA group and simple rapamycin group on the expression of caspase-3 and caspase-8 and the apoptosis rate of 661W cells at different time points (all at P＞0.05).The expressions of Atg-5 and LC-3 Ⅱ/LC-3 Ⅰ in the simple rapamycin group were significantly higher than those in the control group at different time points (all at P＜0.05).The expressions of caspase-3 and caspase-8 and the apoptosis rate in the model + 3-MA group were significantlly higher than those in the model group at 3,6,12,24 and 48 hours after induction,while the expressions of Atg-5 and LC-3 Ⅱ/LC-3 Ⅰ were obviously lower than those in the model group at 3,6,12 and 24 hours after induction (all at P＜0.05).The expressions of caspase-3 and caspase-8 and the apoptosis rate at 6,12,24 and 48 hours after induction in the model+rapamycin group were significantly lower than those in the model group,while the expressions of Atg-5 and LC-3 Ⅱ/LC-3 Ⅰ at the time points of 3,6,12 and 24 hours after induction were obviously higher than those in the model group (all at P ＜ 0.05).Conclusions Under the condition of anti Fas antibody inducing apoptosis,enhancing autophagy can reduce the apoptosis rate of cells,inhibiting autophagy can increase the apoptosis rate.Autophagy may play a protective role in 661W cells.