Objective To investigate the physiological characteristics of Bacillus thuringiensis subspecies israelensis (Bti) with double mutations of cwlE and sigK genes and to assess the activity of Bti against larvae of Culex pipiens pallens under different external factors, so as to provide the theoretical evidence for the use of engineered bacteria of Bti for effective mosquito control. Methods B. thuringiensis wild-type strain Bt-59 and Bt-59 strain with cwlE mutation [Bt-59 (ΔcwlE)] were cultured in nutrient broth media for 24 hours, and Bt-59 strains with sigK mutation [Bt-59 (ΔsigK)] and double mutations of cwlE and sigK [Bt-59 (ΔcwlE-sigK)] were cultured in nutrient broth media for 48 hours. Then, 5 μL of culture media were transferred to glass sides, and cell morphology and mother cell lysis were observed under an optical microscope. The optical densities of Bti strain culture media were measured at different time points of culture, and the growth curves of Bt-59, Bt-59 (ΔcwlE), Bt-59 (ΔsigK), and Bt-59 (ΔcwlE-sigK) strains were plotted. The differences in carbon source metabolism of four Bti strains were analyzed using the Biolog microplate culture method, and the metabolic activity of these strains was estimated with average well color development (AWCD). The fermentation media of these four Bti strains were diluted into final concentrations of 2.000, 1.000, 0.500, 0.250, and 0.125 μL/L, and the median lethal concentrations (LC₅₀ values) of these four strains against the third instar larvae of Cx. pipiens pallens were determined. In addition, the fermentation media of Bti strains were processed as follows: pH adjusted to 5, 7 and 9; treated at 30, 40 ℃ and 50 ℃ for 12 hours; and exposed to irradiation with ultraviolet lights for 0 hour and 6 hours. Then, 20 third instar larvae of Cx. pipiens pallens were exposed to the above processed fermentation media at a final concentration of 1 μL/L in 200 mL of water at 26 ℃ for 24 hours, and the mosquito mortality was estimated to evaluate the effects of pH, temperature and ultraviolet irradiation on the larvicidal activity of four Bti strains. Results The growth curves of the Bt-59 strain and its mutants shared a similar changing trend, and both experienced a stable phase 6 hours post-culture. Both spores and crystal proteins were found in Bt-59 and Bt-59 (ΔcwlE) cells, and only crystal proteins were found in Bt-59 (ΔsigK) and Bt-59 (ΔcwlE-sigK) cells. No lysis was found in the cell wall of the Bt-59 (ΔcwlE-sigK) strain, and the crystal protein was embedded in the mother cell. Biolog microplate culture assay showed that the AWCD values of four Bti strains showed a similar changing trend over time, and 33 carbon sources were found to be metabolized by all of the four strains, including dextrin, D-maltose and D-trehalose. The LC50 values of the fermentation media of Bt-59, Bt-59 (ΔcwlE), Bt-59 (ΔsigK), and Bt-59 (ΔcwlE-sigK) strains were 0.60, 0.51, 0.70 μL/L and 0.72 μL/L against Cx. pipiens pallens, respectively. The adjusted mortality of larval Cx. pipiens pallens reduced by 76.60%, 76.00%, 66.67%, and 0 following exposure to the fermentation media of Bt-59, Bt-59 (ΔcwlE), Bt-59 (ΔsigK), and Bt-59 (ΔcwlE-sigK) strains at a pH of 5 relative to at a pH of 7, and the adjusted mortality reduced by 49.02%, 51.06%, 36.36%, and 4.44% following 6-hour exposure to ultraviolet irradiation relative to 0-hour exposure, while the adjusted mortality was 68.33% to 83.33% following treatment with the fermentation media of four Bti strains at different temperatures. Conclusions Bt-59 (ΔcwlE-sigK) strains do not generate spores, and the absence of cwlE and sigK does not affect the growth, carbon source metabolism, and larvicidal activity of Bti strains against larval Cx. pipiens pallens. Cell wall embedding of Bt-59 (ΔcwlE-sigK) strains may protect larvicidal crystal proteins of Bti strains from external environmental factors, including ultraviolet irradiation, and pH alteration.

ObjectiveTo explore the mechanism of Gancao Fuzitang （GCFZ）in inhibiting the bone destruction of collagen-induced arthritis （CIA） model in mice. MethodThirty male DBa/1J mice were randomly divided into normal group， CIA group， low-dose GCFZ group （GCFZ-L， 2.4 g·kg^-1）， high-dose GCFZ group （GCFZ-H， 4.8 g·kg^-1）， and methotrexate group （MTX， 1 mg·kg^-1）， with six mice in each group. The CIA model was induced by secondary immunization method. The arthritis index of mice in each group was observed and recorded， and the histopathological changes in ankle joint were observed by hematoxylin-eosin （HE） staining. The damage to ankle cartilage was detected by safranin O-fast green staining. Micro-CT scanning was used to detect the bone destruction of ankle joint， and the expression of nuclear factor-κB p65 （NF-κB p65）， p-NF-κB p65， inhibitory-κB kinase α/β （IKKα/β）， and p-IKKα/β was observed by immunohistochemical staining. ResultCompared with the normal group， the CIA group showed manifest joint swelling and increased arthritis index score （P<0.01）. Compared with the CIA group， the groups with drug intervention could inhibit joint swelling and reduce arthritis index score （P<0.05， P<0.01）. As revealed by HE staining and safranine O-green staining， compared with the CIA group， the groups with drug intervention could inhibit synovial invasion and reduce the destruction of articular cartilage. Micro-CT scanning analysis showed that compared with the CIA group， the GCFZ-H group and the MTX group showed reduced bone destruction scores （P<0.01）. The immunohistochemical results showed that compared with the normal group， the CIA group showed increased optical density values of NF-κB p65， p-NF-κB p65， IKKα/β， and p-IKKα/β（P<0.01）. Compared with the CIA group， the GCFZ-H group and the MTX group showed reduced optical density values of NF-κB p65， p-NF-κB p65， IKKα/β， and p-IKKα/β（P<0.05，P<0.01）. In the GCFZ-L group， only the NF-κB p65 optical density value decreased（P<0.01）. ConclusionGCFZ may inhibit bone destruction in CIA mice by regulating the NF-κB signaling pathway.

Objective: To improve the N biomarker in the amyloid/taueurodegeneration system by radiomics and study its value for predicting cognitive progression in individuals with mild cognitive impairment (MCI). Materials and Methods: A group of 147 healthy controls (HCs) (72 male; mean age ± standard deviation, 73.7 ± 6.3 years), 197 patients with MCI (114 male; 72.2 ± 7.1 years), and 128 patients with Alzheimer’s disease (AD) (74 male; 73.7 ± 8.4 years) were included. Optimal A, T, and N biomarkers for discriminating HC and AD were selected using receiver operating characteristic (ROC) curve analysis. A radiomics model containing comprehensive information of the whole cerebral cortex and deep nuclei was established to create a new N biomarker. Cerebrospinal fluid (CSF) biomarkers were evaluated to determine the optimal A or T biomarkers. All MCI patients were followed up until AD conversion or for at least 60 months. The predictive value of A, T, and the radiomics-based N biomarker for cognitive progression of MCI to AD were analyzed using Kaplan-Meier estimates and the log-rank test. Results: The radiomics-based N biomarker showed an ROC curve area of 0.998 for discriminating between AD and HC. CSF Aβ42 and p-tau proteins were identified as the optimal A and T biomarkers, respectively. For MCI patients on the Alzheimer’s continuum, isolated A+ was an indicator of cognitive stability, while abnormalities of T and N, separately or simultaneously, indicated a high risk of progression. For MCI patients with suspected non-Alzheimer’s disease pathophysiology, isolated T+ indicated cognitive stability, while the appearance of the radiomics-based N+ indicated a high risk of progression to AD. Conclusion We proposed a new radiomics-based improved N biomarker that could help identify patients with MCI who are at a higher risk for cognitive progression. In addition, we clarified the value of a single A/T/N biomarker for predicting the cognitive progression of MCI.

Objective:To investigate the efficacy and safety of the modified gasless unilateral axillary approach (MGUAA) endoscopic thyroid surgery in treatment of papillary thyroid microcarcinoma (PTMC) .Methods:From Jan. 2019 to Dec. 2019, 90 patients receiving PTMC (cT1N0M0, cI stage, 8th, 2017 AJCC) therapy by modified gasless unilateral axillary approach endoscopic thyroid surgery (MGUAA group, n=41) , and conventional open thyroid surgery (OS group, n=49) were retrospectively analyzed. Ninety patients were enrolled in the study, including 14 males and 76 females,with the mean age (42.1±12.0) years.The effectiveness of central lymph node dissection (CLND) , the operation time, the types of operation, the amount of drainage, the duration of hospital stay, the related complications, the postoperative pain of neck and axillary and the cosmetic satisfaction were compared between the two groups.SPSS 25.0 statistical software was used for statistical analysis, the measurement data was expressed by ±s, paired t test was used to compare the measurement data between groups, and Chi-square test was used to campare the count date between groups. Results:The mean age (35.0±8.6) years and the amount of surgical bleeding (12.3±7.3) ml in the MGUAA group were significantly lower than those (48.1±11.1) years and (16.1±4.3) ml in the OS group ( P<0.01) , while the mean operation time (99.1±19.5) min, the mean amount of drainage (221.4±67.9) ml and the postoperative drainage tube placement time (5.0±0.8) days were significantly higher than those of (70.6±17.8) min, (98.3±63.7) ml and (3.8±1.0) days in the MGUAA ( P<0.01) . There was no significant difference in the number of lymph nodes of CLND or the duration of hospital stay between the two groups ( P>0.05) . In terms of surgical complications, the transient recurrent laryngeal nerve injury, the postoperative hematoma, the postoperative infection, and the lymphatic leakage had no significant difference between the two groups ( P>0.05) . The MGUAA group had significant advantages in avoiding the postoperative dysphagia in front of neck, the postoperative pain of neck, and cosmetic satisfaction over the OS group [ (0.0% vs 28.6%) , (14.6% vs 71.4%) , (1.1±0.3) score vs (2.4±0.5) score ( P<0.01) ]. Whereas in axillary area pain on the surgical side, the MGUAA group was inferior to the OS group ( P<0.01) . Conclusion:The modified gasless unilateral axillary approach endoscopic thyroid surgery is a feasible, safe and cosmetically operation for PTMC (cT1N0M0, cI stage, 8th, 2017 AJCC) .

Objective:To investigate the effect of treadmill training on spasticity and the expression of potassium chloride co-transporter 2 (KCC2) after blocking BDNF-TrkB signaling pathway in rats with incomplete spinal cord injury (SCI).Methods:Forty female Sprague-Dawley rats were randomly divided into a sham-operation group (Sham group), an SCI+ phosphate-buffered saline group (SCI/PBS group), an SCI-treadmill training+ PBS group (SCI-TT/PBS group), an SCI/TrkB-IgG group and an SCI-TT/TrkB-IgG group. All of the rats underwent 1 week of intrathecal catheterization, and then T 10 incomplete SCI was induced. In the Sham group the spinal cord was only exposed. Seven days later, BDNF-TrkB signaling was blocked in the SCI/TrkB-IgG and SCI-TT/TrkB-IgG groups using the TrkB-IgG. The remaining three groups were controls treated with PBS. The SCI-TT/PBS and SCI-TT/TrkB-IgG groups began exercising 7 days after the SCI and continued for 4 weeks. The spasticity in their hind limbs was assessed using the Asworth assessment and H reflex (H-max/M-max ratio). The expression of KCC2 in the distal spinal cord was detected using western blotting and immunohistochemistry. Results:After the SCI the average Ashworth spasticity grades of the four SCI groups increased significantly compared with the Sham group. The average Ashworth spasticity grade of the SCI-TT/PBS group was significantly lower than those of the SCI/PBS and SCI/TrkB-IgG groups in the 3rd through the 5th week, and the SCI-TT/PBS group′s average grade was significantly less than that of the SCI-TT/TrkB-IgG group after 4 weeks. Within 5 weeks the average H-max/M-max ratio of the Sham group remained unchanged, significantly lower than the other 4 groups′ averages. There was no significant difference in the H-max/M-max ratio among the 4 groups of injured rats within 2 weeks after the SCI, but after 3-5 weeks the average H-max/M-max ratio of the SCI-TT/PBS group was significantly lower than those of the SCI/PBS, SCI/TrkB-IgG and SCI-TT/TrkB-IgG groups. At the 4th and 5th week the average H-max/M-max ratio in the SCI-TT/TrkB-IgG group was significantly lower than that in the SCITrkB-IgG group. And after 5 weeks the average expression of KCC2 in the anterior horn of the injured spinal cord was significantly lower in the 4 SCI groups than in the Sham group. Exercise significantly increased the expression of KCC2 in the SCI-TT/PBS group, and its immune intensity and relative optical density were significantly higher than those in the SCI/PBS, SCI/TrkB-IgG and SCI-TT/TrkB-IgG groups. However, there was no significant difference between the SCI/TrkB-IgG group and the SCI-TT/TrkB-IgG group.Conclusions:Treadmill training can improve spasticity after incomplete SCI and the expression of KCC2 in the distal spinal cord, at least in rats.

Objective To investigate the target blood pressure level of restrictive fluid resuscitation in patients with traumatic hemorrhagic shock. Methods Sixty patients with traumatic hemorrhagic shock admitted to the First Affiliated Hospital of Bengbu Medical College from January 2016 to December 2018 were enrolled. All patients were resuscitated with sodium acetate ringer solution after admission. According to the difference of mean arterial pressure (MAP) target, the patients were divided into low MAP (60 mmHg ≤ MAP < 65 mmHg, 1 mmHg = 0.133 kPa), middle MAP (65 mmHg ≤ MAP < 70 mmHg) and high MAP (70 mmHg ≤ MAP < 75 mmHg) groups by random number table using the admission order with 20 patients in each group. Those who failed to reach the target MAP after 30-minute resuscitation were excluded and supplementary cases were deferred. The restrictive fluid resuscitation phase was divided into three phases: before fluid resuscitation, liquid resuscitation for 30 minutes and 60 minutes. The most suitable resuscitation blood pressure level was further speculated by monitoring the inflammatory markers and hemodynamics in different periods in each group of patients. Pearson correlation analysis was used to detect the correlation of variables. Results Before fluid resuscitation, there was no significant difference in hemodynamics or expressions of serum cytokines among the three groups. Three groups of patients were resuscitated for 30 minutes to achieve the target blood pressure level and maintain 30 minutes. With the prolongation of fluid resuscitation time, the central venous pressure (CVP), cardiac output (CO) and cardiac index (CI) were increased slowly in the three groups, and reached a steady state at about 30 minutes after resuscitation, especially in the high MAP group and the middle MAP group. The expressions of serum inflammatory factors in the three groups were gradually increased with the prolongation of fluid resuscitation time. Compared with the low MAP group and the high MAP group, after 30 minutes of resuscitation the middle MAP group was superior to the other two groups in inhibiting the expressions of pro-inflammatory factors tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and promoting anti-inflammatory factors IL-10 [TNF-α mRNA (2-ΔΔCt):0.21±0.13 vs. 0.69±0.34, 0.57±0.35; IL-6 mRNA (2-ΔΔCt): 0.35±0.31 vs. 0.72±0.39, 0.59±0.42; IL-10 mRNA (2-ΔΔCt): 1.25±0.81 vs. 0.61±0.46, 0.82±0.53; all P < 0.05], but there was no significant difference in promoting the expression of IL-4 mRNA among three groups. At 60 minutes of resuscitation, compared with the low MAP group and the high MAP group, the middle MAP group could significantly inhibit the expressions of TNF-α, IL-6 and promote IL-10 [TNF-α mRNA (2-ΔΔCt): 0.72±0.35 vs. 1.05±0.54, 1.03±0.49; IL-6 mRNA (2-ΔΔCt): 0.57±0.50 vs. 1.27±0.72, 1.01±0.64; IL-10 mRNA (2-ΔΔCt): 1.41±0.90 vs. 0.81±0.48, 0.94±0.61; all P < 0.05]. Compared with the high MAP group, the middle MAP group had significant differences in promoting the expression of IL-4 mRNA (2-ΔΔCt: 1.32±0.62 vs. 0.91±0.60, P < 0.05). There was no significant difference in serum cytokine expressions at different time points of resuscitation between the low MAP group and the high MAP group (all P > 0.05). Correlation analysis showed that there was a strong linear correlation between MAP and mRNA expressions of TNF-α, IL-6, IL-10 in the middle MAP group (r value was 0.766, 0.719, 0.692, respectively, all P < 0.01), but had no correlation with IL-4 (r = 0.361, P = 0.059). Fitting linear regression analysis showed an increase in 1 mmHg per MAP, the expression of TNF-α mRNA increased by 0.027 [95% confidence interval (95%CI) = 0.023-0.031, P < 0.001], IL-6 mRNA increased by 0.021 (95%CI = 0.017-0.024, P < 0.001), and IL-10 mRNA increased by 0.049 (95%CI = 0.041-0.058, P < 0.001). Conclusions When patients with traumatic hemorrhagic shock received restrict fluid resuscitation at MAP of 65-70 mmHg, the effect of reducing systemic inflammatory response and improving hemodynamics is better than the target MAP at 60-65 mmHg or 70-75 mmHg. It is suggested that 65-70 mmHg may be an ideal target MAP level for restrictive fluid resuscitation.

Objective To evaluate the potential cerebral cortical volume alterations in type 2 diabetes mellitus (T2DM) patients with mild cognitive impairment (MCI) compared with T2DM patients and healthy controls,and to observe the correlations with the scores of neuropsychological scales.Methods Cortical volume based on high-resolution MR T1WI data from 30 healthy controls (HC),30 T2DM patients and 30 T2DM with MCI patients were evaluated with FreeSurfer software and compared with variance analysis.The correlations between cerebral cortical volume which had statistical difference and the scores of neuropsychological scales were analyzed.Results There were significant differences in auditory verbal learning test (AVLT) scores,complex figure test-delayed recall (20 min) scores,digit symbol-coding subtest scores,MoCA scores and higher trail-making test-A scores,as well as trail-making test-B scores between T2DM and T2DM with MCI patients (all P＜0.05).Compared with T2DM patients,cortical volume of left entorhinal cortex,left lateral orbitofrontal gyrus,left posterior cingulate gyrus and the right lateral orbitofrontal gyrus,right pars orbitalis,right insula reduced in T2DM with MCI patients (all P＜0.05).In T2DM with MCI patients,AVLT scores were positively correlated with volume of the left entorhinal cortex (r=0.452,P=0.018).Conclusion Several cortical volume reductions are exhibited in T2DM patients with MCI.The volume of the left entorhinal cortex may be a potential biomarker to diagnose and evaluate MCI in T2DM.

Objective To analyze the clinical values of super early enteral nutrition combined with microecopharmaceutics and delayed enteral nutrition on patients with severe acute pancreatitis. Methods Clinical data of thirty patients diagnosed as severe acute pancreatitis in our emergency department during January 2013 and December 2017 were reviewed retrospectively. Patients were divided into the treatment group (n=15, patients given enteral nutrition combined with microecopharmaceutics within 24 h after admission) and the control group (n=15, patients given delayed enteral nutrition after 48 h of admission). Two weeks after the treatment, the serum variables of C-reactive protein, total protein, albumin, recovery time of urine and blood amylase, length of hospital stay and APACHE Ⅱ score were compared between the two groups by using paired samples t test. Results The C-reactive protein [(46.7±13.1) mg/L vs. (190.72±19.3) mg/L, t=10.4, P<0.01] and APACHE Ⅱ score [(7.2±1.9) vs.(9.3±2.4),t=2.7,P<0.05] of the treatment group were significantly lower than those in the control group. The total protein [(58.1±6.3)g/L vs.(52.6±5.4)g/L, t=2.5, P<0.05] and albumin [(29.9±3.2)g/L vs.(22.0±2.8)g/L, t=7.12, P<0.01] of the treatment group were significantly higher than those in the control group. The recovery time of urine amylase [(13.2±2.1)d vs.(18.7±3.9)d, t=4.9, P<0.01] and blood amylase [(7.5±3.0)d vs.(11.1±3.4)d, t=3.1, P<0.01], and length of hospital stay[(14.9±4.5)d vs.(27.1±5.3)d, t=6.9, P<0.01] were significantly shorter in the treatment group compared with those in the control group. Conclusions Ultra-early enteral nutrition combined with microecopharmaceutics can shorten the length of hospital stay of patients with severe acute pancreatitis, and is safe and effective.

Objective: To investigate the effect of icariin (ICA) on the bisphenol A (BPA)-enhanced proliferation function of thyroid carcinoma cell B-CPAP and underlying mechanism. Methods: The proliferation of Gastric B-CPAP cell line was evaluated by cell counting kit-8 (CCK-8). Apoptosis and ROS expression in B-CPAP cells were detected by flow cytometry. The expression of superoxide dismutase (SOD) and malondialdehyde (MDA) in B-CPAP cells were measured by individual assay kits. The expressions of Bcl-2 and γ-HA2X were detected by Western blot. SPSS 18.0 software was used to analyze the data. Results: B-CPAP cell activity was promoted by treatment with 3×10^-7mol/L BPA for 48 h, with significant difference in absorbance between BPA and control groups (1.089±0.053 vs 0.935±0.010, P<0.05). The cell activities of BPA+ ICA₂₅, BPA+ ICA₅₀, BPA+ ICA₁₀₀ and BPA+ ICA₂₀₀ groups was 0.780±0.036, 1.007±0.050, 0.958±0.033 and 0.625±0.064, respectively (all P<0.01). The proliferation of B-CPAP cells treated with BPA for 72 hours showed a similar trend to 48 hours. There was no significant difference between all treatment groups in 24 hours. The apoptosis rate was (19.272±0.186)% in BPA-treated cells, and was (22.412±0.238)% in control cells (P<0.05). The apoptosis rates of BPA+ ICA₅₀ and BPA+ ICA₂₀₀ groups were (23.688±0.412)% and (30.270±0.696)%, respectively (P<0.01). The intracellular accumulation of ROS in BPA, BPA+ ICA₅₀, and BPA+ ICA₂₀₀ groups were 806±21, 1 772±37, 2 041±16, respectively (P<0.01). The expressions of anti-apoptotic protein Bcl-2 in control, BPA, BPA+ ICA₅₀, BPA+ ICA₂₀₀ groups were 7 120±151, 9 801±286, 5 902±171 and 4 203±216, respectively (P<0.01). Conclusion BPA can promote the proliferation of thyroid carcinoma B-CPAP cells and decrease the apoptosis of cells, and this effect can be inhibited by ICA. The possible mechanism is to induce high expression of intracellular ROS and inhibit the expression of antioxidase system, leading to cell oxidative damage, thereby inducing apoptosis.

Objective To explore CT performance of primary pulmonary lymphoma(PPL)and analyze the cause of misdiagnosis. Methods CT data of 42 cases with PPL which confirmed by pathology and clinical were analyzed.All cases met the diagnostic criteria of Kim et al[1] and CT analysis of lesions included the location,shape,border,enhanced degree and internal characteristic,etc.CT images of 29 preoperative misdiagnosis cases were also analyzed and summarized.Results 34 cases underwent both pre-contrast and post-contrast scan and 8 cases only had plain CT scan.The cases included mass or nodular type(21 cases),pneumonic-alveolar type(11 cases), bronchovascular-lymphatic type(4 cases),and mixed pattern(6 cases).Meanwhile,CT features contained air-bronchus sign (32 cases),angiogram sign(20 cases),airway dilatation(4 cases)and cross lobe growing(4 cases).29 cases were misdiagnosed as other lung diseases,thus misdiagnosis rate accounted for 69%.Conclusion PPLs mainly present with intrapulmonary nodules, masses and patches.In the lesions,air-bronchus sign or angiogram sign can be seen,and the density of solid portion is homogeneous with mildly to moderately homogeneous enhancement.The coexistence of multiple-type lesions and trans-lobes distribution have certain characteristics.