To analyze the distribution of serotypes and antimicrobial resistance of clinical Salmonella isolates from multiple centers across China.

Objective To explore the effects of different grinding amounts of articular processes on the biomechanical stability and inflammatory response of lumbar spine during percutaneous foraminoscopic surgery(PTED).Methods A total of 195 patients with lumbar disc herniation(LDH)in our hospital were selected for prospective study from January 2018 to October 2020,and they were divided into three groups by randomized numerical table method,each with 65 cases,and all of them were implemented PTED.Patients with intraoperative upper joint process grinding＜33％were treated as a small group,patients with intraoperative grinding＜33％～50％were treated as a medium group,and patients with＞50％were treated as a large group.The surgery-related indexes of the three groups of patients were observed,as well as the inflammatory indexes[interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α)],biomechanical stability,and joint functional recovery scores at different time points before and after surgery.Results The operation time and intraoperative bleeding were(61.32±7.86)min and(11.85±1.10)ml in the small group,(70.06±8.53)min and(14.32±2.21)ml in the medium group,and(74.47±10.00)min and(19.86±3.00)ml in the large group,and the differences among the three groups were statistically significant(P＜0.05).Serum IL-6 and TNF-α were(6.10±1.12)pg/ml and(8.21±1.87)ng/L in the small group 3d postoperatively,and(3.27±0.58)pg/ml and(4.32±1.00)ng/L in the discharge group,and in the medium group 3d postoperatively,they were(6.68±1.35)pg/ml and(9.00±1.20)ng/L at discharge and(4.10±0.63)pg/ml and(6.85±1.28)ng/L at discharge,respectively,and(7.32±1.00)pg/ml and(10.57±1.28)ng/L in the massive group at 3d postoperatively and(4.57±0.49)pg/ml and(8.14±1.35)ng/L at discharge,and the difference between the three groups was statistically significant(P＜0.05);the serum lumbar lordosis angle,lumbar flexion,and lumbar stability scores were(42.28±2.12)°,(1.86±0.36)cm,and(10.52±2.01)° at 1 month postoperatively,and(42.11±1.97)°,(1.87±0.52)cm,and(10.63±1.96)° at 1 year postoperatively,respectively,for the small group,and(40.86±1.89)°,(1.72±0.28)cm,(10.63±2.15)scores,(39.15±2.11)°,(1.60±0.42)cm,(12.11±1.63)scores for the medium group,respectively,1 month postoperatively in the large volume group,(39.01±2.35)°,(1.61±0.29)cm,(11.20±2.75)scores,and(38.11±2.32)°,(1.43±0.33)cm,(13.24±1.52)scores at 1 year postoperatively,respectively,and the difference between the three groups were statistically significant(P＜0.05).Conclusion PTED treatment of LDH can reduce the amount of grinding of the superior articular process during surgery,reduce operative time and intraoperative bleeding,alleviate inflammatory response,and achieve good short-and medium-term lumbar function and joint stability.

This study established the mouse podocyte clone-5 (MPC5) with Smad3 knockout and studied the effect of transforming growth factor-beta 1 (TGF-β1) on the dedifferentiation of the MPC5 cells with Smad3 knockout, aiming to provide a cell tool for studying the role of Smad3 in mouse podocytes. The single-guide RNA (sgRNA) sequence targeting Smad3 was designed according to the principles of CRISPR/Cas9 design. The pX458-Smad3 vector was constructed and introduced into competent cells, and then the vector was extracted and used to transfect MPC5 cells. The successfully transfected cells were sorted by a flow cytometer. After single-cell clone expansion, PCR amplification of sequences adjacent to the edition site of Smad3 and sequencing were performed to identify potential cells with gene knockout. Western blotting was employed to verify the knockout efficiency of Smad3. Finally, the effect of Smad3 knockout on TGF-β1-induced dedifferentiation of MPC5 cells was analyzed by reverse transcription-polymerase chain reacting (RT-PCR), Western blotting, and the immunofluorescence method. The sgRNA was designed to target the fifth exon of Smad3. EGFP expression was observed 24 h after transfection of the pX458-Smad3 plasmid into MPC5 cells, with the transfection efficiency of 0.1% as determined by flow cytometry. From the transfected cells, 21 cell clones were obtained through flow cytometric sorting and single-cell clone expansion. PCR amplification and sequencing of the region around the sgRNA target site in Smad3 identified two cell clones with biallelic frameshift mutations. Western blotting results confirmed the absence of Smad3 expression in these clones, indicating successful establishment of the MPC5 cell line with Smad3 knockout. In normal MPC5 cells, TGF-β1 stimulation promoted the expression of fibrosis-related genes fibronectin and Col1a1 (collagen I) and inhibited the expression of the podocyte marker proteins synaptopodin and podocin, which suggested epithelial-mesenchymal transition and podocyte injury. However, in the two MPC5 cell lines with Smad3 knockout, TGF-β1-induced expression of epithelial-mesenchymal transition markers was significantly suppressed. The MPC5 cell lines with Smad3 knockout that were constructed by CRISPR/Cas9 provide a valuable cell model for functional studies of Smad3 protein and highlight the critical role of Smad3 in cell dedifferentiation.
Animals ; Smad3 Protein/genetics* ; CRISPR-Cas Systems/genetics* ; Mice ; Podocytes/metabolism* ; Transforming Growth Factor beta1/pharmacology* ; Cell Line ; Gene Knockout Techniques ; RNA, Guide, CRISPR-Cas Systems/genetics*

Objective To explore the effects of different grinding amounts of articular processes on the biomechanical stability and inflammatory response of lumbar spine during percutaneous foraminoscopic surgery(PTED).Methods A total of 195 patients with lumbar disc herniation(LDH)in our hospital were selected for prospective study from January 2018 to October 2020,and they were divided into three groups by randomized numerical table method,each with 65 cases,and all of them were implemented PTED.Patients with intraoperative upper joint process grinding＜33％were treated as a small group,patients with intraoperative grinding＜33％～50％were treated as a medium group,and patients with＞50％were treated as a large group.The surgery-related indexes of the three groups of patients were observed,as well as the inflammatory indexes[interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α)],biomechanical stability,and joint functional recovery scores at different time points before and after surgery.Results The operation time and intraoperative bleeding were(61.32±7.86)min and(11.85±1.10)ml in the small group,(70.06±8.53)min and(14.32±2.21)ml in the medium group,and(74.47±10.00)min and(19.86±3.00)ml in the large group,and the differences among the three groups were statistically significant(P＜0.05).Serum IL-6 and TNF-α were(6.10±1.12)pg/ml and(8.21±1.87)ng/L in the small group 3d postoperatively,and(3.27±0.58)pg/ml and(4.32±1.00)ng/L in the discharge group,and in the medium group 3d postoperatively,they were(6.68±1.35)pg/ml and(9.00±1.20)ng/L at discharge and(4.10±0.63)pg/ml and(6.85±1.28)ng/L at discharge,respectively,and(7.32±1.00)pg/ml and(10.57±1.28)ng/L in the massive group at 3d postoperatively and(4.57±0.49)pg/ml and(8.14±1.35)ng/L at discharge,and the difference between the three groups was statistically significant(P＜0.05);the serum lumbar lordosis angle,lumbar flexion,and lumbar stability scores were(42.28±2.12)°,(1.86±0.36)cm,and(10.52±2.01)° at 1 month postoperatively,and(42.11±1.97)°,(1.87±0.52)cm,and(10.63±1.96)° at 1 year postoperatively,respectively,for the small group,and(40.86±1.89)°,(1.72±0.28)cm,(10.63±2.15)scores,(39.15±2.11)°,(1.60±0.42)cm,(12.11±1.63)scores for the medium group,respectively,1 month postoperatively in the large volume group,(39.01±2.35)°,(1.61±0.29)cm,(11.20±2.75)scores,and(38.11±2.32)°,(1.43±0.33)cm,(13.24±1.52)scores at 1 year postoperatively,respectively,and the difference between the three groups were statistically significant(P＜0.05).Conclusion PTED treatment of LDH can reduce the amount of grinding of the superior articular process during surgery,reduce operative time and intraoperative bleeding,alleviate inflammatory response,and achieve good short-and medium-term lumbar function and joint stability.

Objective To study the initial pumping speed of sodium citrate in single plasma exchange with regional citrate anticoagulation(RAC).Methods From January to December 2021,15 patients and 67 times of treatment with local sodium citrate anticoagulation single plasma exchange in the hospital were in-cluded in the study.According to the initial pumping speed of sodium citrate,they were included in the low-speed group(n=33)and the high-speed group(n=34).The transmembrane pressure,filter pressure drop and venous pressure were compared between the two groups at 30 minutes,one hour and two hours after treatment.The free calcium concentration after plasma separator at 15 minutes and one hour after treatment,and the coagulation of plasma separator and extracorporeal circulation pipeline at the end of treatment were compared between the two groups.The concentration of free calcium,blood gas analysis and electrolyte were compared at the beginning of treatment,one hour after treatment and at the end of treatment.Results The free calcium concentration after the filter was monitored at 15 minutes and one hour of treatment in both groups was within the effective range of anticoagulation recommended by the guidelines.There were no lips,fingertip numbness and hand-foot convulsions in the two groups during the treatment,and no bleeding oc-curred after the treatment.There were four cases of hypocalcemia and two cases of alkalosis in the low-speed group,and 13 cases of hypocalcemia and eight cases of alkalosis in the high-speed group.The difference be-tween the two groups was statistically significant(P＜0.05).There were 15 cases of grade Ⅰ coagulation and five cases of grade Ⅱ coagulation in plasma separator and pipeline in the low-speed group,while there were 14 cases of grade Ⅰ coagulation and four cases of grade Ⅱ coagulation in plasma separator and pipeline in the high-speed group.There was no significant difference between the two groups(P＞0.05).Conclusion In plasma exchange treatment,according to the low initial pumping speed,RAC can not only ensure the anticoagulant effect,but also reduce the incidence of complications such as hypocalcemia and alkalosis.

Objective:To explore the predictive value of uterine artery ultrasound parameters combined with four coagulation indicators test for placental abruption in late pregnancy.Methods:A total of 160 pregnant women with placental abruption during the late pregnancy who were diagnosed and treated in our hospital from Jan. 2021 to Dec. 2022 were collected as the observation group, and 160 pregnant women with normal delivery were regarded as the control group. According to different grading of placental abruption, 65 cases were classified as grade I, 54 cases as grade II, and 41 cases as grade III. Ultrasound parameters of uterine artery [uterine artery pulsatility index (PI), resistance index (RI), ratio of maximum systolic blood flow velocity (S) to maximum diastolic blood flow velocity (D) (S/D) ], and four coagulation parameters [prothrombin time (PT), fibrinogen (FIB), thrombin time (TT), and activated partial thromboplastin time (APTT) ] were compared; Logistic regression analysis was conducted to analyze the factors affecting placental abruption in late pregnancy; receiver operating characteristic (ROC) curve was applied to analyze the predictive value of uterine artery ultrasound parameters combined with four coagulation indicators test for placental abruption in late pregnancy.Results:Compared with the control group, PI, RI, and S/D ratios of pregnant women with placental abruption in the late pregnancy in the observation group were obviously increased [ (1.26 ± 0.22) vs. (0.95 ± 0.14), (0.65 ± 0.12) vs. (0.48 ± 0.06), (3.46 ± 0.63) vs. (2.57 ± 0.45) ] (P<0.05) ; compared with the control group, PT, TT, and APTT in pregnant women with placental abruption in the late pregnancy in the observation group were obviously increased [ (12.90 ± 1.42) vs. (10.24 ± 1.14), (15.06 ± 1.24) vs. (12.67 ± 1.08), (30.32 ± 2.55) vs. (25.48 ± 2.10) ] ( P<0.05), the FIB level was obviously decreased [ (3.09 ± 0.37) g/L vs. (3.96 ± 0.58) g/L] (P<0.05) ; compared with the grade I group, as the grading of placental abruption increased, PI, RI, S/D ratio, PT, TT, and APTT in grade II group and grade III group increased in turn ( P<0.05) and the level of FIB decreased in turn ( P<0.05) ; Logistic regression analysis showed that mechanical injury, polyhydramnios, premature rupture of membranes, pregnancy diabetes, pregnancy induced hypertension, PI, RI, S/D ratio, PT, TT, APTT were the risk factors for placental abruption in late pregnancy ( P<0.05), and FIB was the protective factor for placental abruption in late pregnancy ( P<0.05) ; the area under the ROC curve (AUC) the combined detection of PI, RI, S/D ratio, PT, FIB, TT, and APTT for placental abruption in late pregnancy was 0.982, which was better than their individual predictions (Z combined test-PI=6.118, P<0.001; Z combined test-RI=6.080, P<0.001; Z combined test-S/D ratio=6.690, P<0.001; Z combined test-PT=5.837, P<0.001; Z combined test-FIB=6.500, P<0.001; Z combined test-TT=6.439, P<0.001; Z combined test- PTT=6.112, P<0.001) . Conclusions:The PI, RI, S/D ratio, PT, TT, APTT increase, and FIB level decreases in pregnant women with placental abruption in the late pregnancy, they are related to different grades of placental abruption. Combined detection has good predictive value for placental abruption in pregnant women in the late pregnancy.

Objective:To analyze the predictive value of combined insulin autoantibody (IAA), islet cell antibody (ICA), and serum glutamic acid decarboxylase antibody (GADA) testing for pregnancy outcome in patients with gestational diabetes mellitus (GDM) .Methods:One hundred and twenty patients with GDM were retrospectively selected for the study, and were divided into a poor pregnancy outcome group and a normal pregnancy outcome group based on the pregnancy outcome at follow-up to delivery, which were 37 and 83 cases, respectively. The occurrence of adverse pregnancy outcomes in the study population was counted, and univariate and multivariate analyses of adverse pregnancy outcomes in GDM were performed.Results:Adverse pregnancy outcomes were seen in 37 of the 120 study subjects, including 12, 13 and 9 cases of macrosomia, caesarean section and neonatal hypoglycaemia, respectively, and 1 case of premature rupture of the foetal hairs, placenta previa and neonatal asphyxia. The percentage of IAA, ICA, and GADA positivity was higher in the adverse pregnancy outcome group than in the normal pregnancy outcome group ( P<0.05). Multifactorial regression analysis showed that IAA ( OR=3.180, 95% CI 1.394-7.258), ICA ( OR=3.459, 95% CI 1.592-7.517), and GADA ( OR=3.219, 95% CI 1.508-6.872) positivity were independent risk factors for adverse pregnancy outcomes in GDM ( P<0.05). The AUC values for the combined IAA, ICA, and GADA for the detection of adverse pregnancy outcomes in GDM were higher than those for IAA, ICA, and GADA alone ( Z=2.607, 2.600, and 2.527, P<0.05) . Conclusion:Positive IAA, ICA, and GADA are risk factors for adverse pregnancy outcomes in patients with GDM, and combined testing has a higher predictive value.

Objective:To analyze the predictive value of combined insulin autoantibody (IAA), islet cell antibody (ICA), and serum glutamic acid decarboxylase antibody (GADA) testing for pregnancy outcome in patients with gestational diabetes mellitus (GDM) .Methods:One hundred and twenty patients with GDM were retrospectively selected for the study, and were divided into a poor pregnancy outcome group and a normal pregnancy outcome group based on the pregnancy outcome at follow-up to delivery, which were 37 and 83 cases, respectively. The occurrence of adverse pregnancy outcomes in the study population was counted, and univariate and multivariate analyses of adverse pregnancy outcomes in GDM were performed.Results:Adverse pregnancy outcomes were seen in 37 of the 120 study subjects, including 12, 13 and 9 cases of macrosomia, caesarean section and neonatal hypoglycaemia, respectively, and 1 case of premature rupture of the foetal hairs, placenta previa and neonatal asphyxia. The percentage of IAA, ICA, and GADA positivity was higher in the adverse pregnancy outcome group than in the normal pregnancy outcome group ( P<0.05). Multifactorial regression analysis showed that IAA ( OR=3.180, 95% CI 1.394-7.258), ICA ( OR=3.459, 95% CI 1.592-7.517), and GADA ( OR=3.219, 95% CI 1.508-6.872) positivity were independent risk factors for adverse pregnancy outcomes in GDM ( P<0.05). The AUC values for the combined IAA, ICA, and GADA for the detection of adverse pregnancy outcomes in GDM were higher than those for IAA, ICA, and GADA alone ( Z=2.607, 2.600, and 2.527, P<0.05) . Conclusion:Positive IAA, ICA, and GADA are risk factors for adverse pregnancy outcomes in patients with GDM, and combined testing has a higher predictive value.

Objective:To analyze the fail mode of neoadjuvant therapy combined with surgery for locally advanced esophageal squamous cell carcinoma (ESCC) after long-term follow-up.Methods:Clinical data of consecutive 238 patients with locally advanced resectable ESCC who underwent neoadjuvant therapy combined with surgery in Zhejiang Cancer Hospital from September 2012 to October 2019 were retrospectively analyzed. The failure mode in the whole cohort was analyzed after long-term follow-up. The overall survival (OS) and disease free survival (DFS) rates were analyzed by Kaplan-Meier method. Survival differences were determined by log-rank test.Results:The pathological complete response (pCR) rate was 42.0% in 238 patients. After a median follow-up of 46.1 months, tumor progression occurred in 96 patients (40.3%), including 25 patients (10.5%) with local recurrence, 61 patients (25.6%) with distant metastases, and 10 patients (4.2%) with simultaneous local recurrence and distant metastases. The median OS and DFS were 64.7 months and 49.9 months. And the 3-, 5-, and 7-year OS and DFS rates were 70.0%, 52.8%, 36.4% and 63.5%, 42.5%, and 30.0%, respectively. The 3-, 5-, and 7-year locoregional recurrence-free survival rates and distant metastasis-free survival rates were 86.0%, 71.4%, 61.2% and 70.6%, 55.9%, 43.0%. Compared with non-pCR patients, the overall progression rate and distant metastasis rate of pCR patients were lower (26.0% vs. 50.7%, 16.0% vs. 32.6%, both P<0.05). And the 3-, 5-, and 7-year OS (83.0% vs. 60.2%, 69.7% vs. 41.7%, 50.4% vs. 27.7%, all P<0.001) and DFS rates (80.4% vs. 51.4%, 63.9% vs. 31.2%, 45.9% vs. 20.3%, all P<0.001) were significantly better in pCR patients. Conclusions:Distant metastasis is the main failure mode of patients with locally advanced ESCC after neoadjuvant therapy. Patients with postoperative pCR can achieve better long-term survival.

Objective:To analyze the failure patterns and survival after stereotactic body radiotherapy (SBRT) in patients with T 1-2N 0M 0 non-small cell lung carcinoma (NSCLC). Methods:Clinical data of early-stage NSCLC patients who received SBRT at Zhejiang Cancer Hospital from January 2012 to September 2018 were retrospectively analyzed. The primary observed endpoint was the pattern of disease progression, which was divided into intra-field recurrence, regional lymph node recurrence and distant metastasis. Overall survival (OS) and progression-free survival (PFS) were calculated by Kaplan-Meier method. Univariate analysis was conducted by log-rank test, and multivariate analysis was performed by Cox's model.Results:A total of 147 patients with 156 lesions were included. The median follow-up time was 44.0 months (16.5-95.5 months). A total of 57 patients (38.8%) progressed: 14 patients (24.5%) had recurrence with the 1-, 3-, and 5-year local recurrence rates of 2.0%, 10.9%, and 14.3%, respectively; 36 patients (63.2%) had Distant metastasis with the 1-, 3- and 5-year distant metastasis rates of 12.2%, 22.4% and 28.6%, respectively; and 7 patients (12.3%) had recurrence complicated with distant metastasis. The 3-, 5- and 7-year OS rates were 80.5%, 64.2% and 49.9% for all patients, respectively. The median OS was 78.4 months. The 3-, 5- and 7-year PFS rates were 64.8%,49.5% and 41.5%, with a median PFS of 57.9 months (95% CI: 42.3-73.5 months). Univariate and multivariate analyses showed that biologically equivalent dose and age were the factors affecting the efficacy of SBRT (both P<0.05). Conclusion:Distant metastasis is the main failure pattern in patients with T 1-2N 0M 0 NSCLC after SBRT. High-risk population should be selected for further systematic treatment to improve the efficacy.