Objective:To evaluate the effect of Lactobacillus-derived extracellular vesicles (Lac-EVs) on lipopolysaccharide (LPS)-induced activation of microglia and proteomic analysis.Methods:BV2 microglia obtained from mice with good growth status were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), LPS group (group L) and LPS+ Lac-EVs group (group L+ E). Group C was commonly cultured. Group L was incubated for 24 h with LPS (final concentration 1 μg/ml). Group L+ E was incubated for 24 h with Lac-EVs (final concentration 2.5 μg/ml) after being treated with LPS for 24 h. The expression of CD86 and CD206 was detected using immunofluorescence staining. Cell precipitates were taken from L and L+ E groups, and proteomics were used to screen for differentially expressed proteins between the two groups. The differentially expressed proteins were analyzed by the bioinformatics analysis, and two differentially expressed proteins, apolipoprotein A1 and G protein-coupled receptor kinase 2, were verified by quantitative real-time polymerase chain reaction and Western blot. Results:Compared with group C, the expression of CD86 was significantly up-regulated, and the expression of CD206 was down-regulated in group L ( P<0.05). Compared with group L, the expression of CD86 was significantly down-regulated, and the expression of CD206 was up-regulated in L+ E group ( P<0.05). One hundred and twenty-five differentially expressed proteins were identified using proteomics (FC=2.0, P<0.05), of which the expression of 66 proteins was up-regulated and the expression of 59 proteins was down-regulated. The results of GO analysis indicated that these differentially expressed proteins were mainly involved in biological processes such as endothelial cell proliferation, SDNA damage detection, and lipoprotein transport. The results of KEGG analysis indicated that there were differences in PPAR signaling pathway, endocytosis, metabolic pathway, MAPK signaling pathway, etc. The expression trends of the differentially expressed proteins determined by Western blot and quantitative real-time polymerase chain reaction were consistent with the results of proteomics. Conclusions:Lac-EVs can inhibit LPS-induced microglial polarization toward M1 phenotype, and the mechanism may be related to the up-regulated differential proteins apolipoprotein A1 and G protein-coupled receptor kinase 2.

Objective:To investigate the application value of antegrade splenic superior region dissection first in laparoscopic total gastrectomy of obesity gastric cancer.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 21 obesity patients with gastric cancer who underwent laparoscopic total gastrectomy in Renji Hospital of Shanghai Jiaotong University School of Medicine from July 2018 to October 2023 were collected. There were 16 males and 5 females, aged (58±13)years. All 21 patients underwent laparoscopic total gastrec-tomy with antegrade splenic superior region dissection first. Observation indicators: operation time, volume of intraoperative blood loss, laparotomy conversion, intraoperative splenic hemorrhage or gastric hemorrhage, lymph node dissection, time to postoperative first flatus, time to postoperative initial liquid food intake, duration of postoperative hospital stay, postoperative complication. Measure-ment data with normal distribution were represented as Mean± SD, and count data were expressed as absolute numbers. Results:All 21 patients underwent laparoscopic total gastrectomy success-fully, with the operation time of (283±47)minutes, time for splenogastric ligament and vascular manage-ment of (34±12)minutes, volume of intraoperative blood loss of (143±86)mL, and no laparotomy conversion. There was no intraoperative splenic hemorrhage or gastric haemorrhage. The total number of lymph node dissected in 21 patients was 375, with the number of lymph node dissected as (21±9)per case. Time to postoperative first flatus, time to postoperative initial liquid food intake and duration of postoperative hospital stay in 21 patients were (3.1±0.7)days, (4.0±0.8)days and (10.1±3.0)days, respectively. There were 2 patients with postoperative complications, including 1 case of incision infection and 1 case of lung infection. The 2 patients with postoperative com-plications were recovered and discharged after conservative treatment. There was no death during the postoperative 30 days.Conclusion:The application of antegrade splenic superior region dissec-tion first in laparoscopic total gastrectomy is safe and feasible, which can reduce surgical difficulty.

Chloroquine has played a pivotal role in the treatment of malaria.Recently,with the development of clinical application,they are increasingly recognized for their effectiveness in other as-pects,such as anti-inflammation,immunosuppression,anti-amoebic effects and so on.In the outbreak of the new coronavirus infection,chloroquine and its derivative hydroxychloroquine swiftly included in the treatment of infected patients in many countries.However,several studies have shown these drugs have some detrimental influence,especially in cardiac arrhythmia(prolonged the QT interval and in-creased risk of torsional ventricular tachycardia).Thus,we should pay attention to both their benefi-cial effects and potential adverse manifestation.In order to regulate the use of drugs and guide the clin-ic,this paper reviewed the electrophysiological basis and clinical characteristics of cardiac adverse re-actions of these drugs from the perspective of ion channels.

OBJECTIVE To develop a method for simultaneous content determination of menthone, menthol, 2-methoxy-4-vinylphenol, 3-butenyl phthalide, atractyline, ligustilide in Honghua Xiaoyao tablets. METHODS The quantitative analysis was carried out on a capillary of HP-5(30 m×250 μm, 0.25 μm) column combined with an oven temperature program; inlet port temperature 250 ℃; detector temperature 280 ℃, split sampling at a split ratio 10∶1. RESULTS The 6 components were completely separated and could be well separated from other components; menthone, menthol, 2-methoxy-4-vinylphenol, 3-butenyl phthalide, atractyline, ligustilide showed a good linear relationship with the chromatographic peak area within the range of 1.37-87.38, 13.67-874.55, 2.05-65.49, 3.35-107.20, 3.49-111.60, 6.95-444.60 μg·mL-1(r≥0.999 0). The average recovery was 97.63%(RSD=3.27%), 94.25%(RSD=1.80%), 97.54%(RSD=2.39%), 103.81%(RSD=1.75%), 94.30% (RSD=2.32%), 98.97%(RSD=2.36%)(n=6), respectively. And its content in 39 batches of Honghua Xiaoyao tablets were determined. CONCLUSION This method is easy to operate, accurate, reliable, and has good repeatability, can be used to supplement the insufficient quality control of Honghua Xiaoyao tablets.

Chloroquine has played a pivotal role in the treatment of malaria.Recently,with the development of clinical application,they are increasingly recognized for their effectiveness in other as-pects,such as anti-inflammation,immunosuppression,anti-amoebic effects and so on.In the outbreak of the new coronavirus infection,chloroquine and its derivative hydroxychloroquine swiftly included in the treatment of infected patients in many countries.However,several studies have shown these drugs have some detrimental influence,especially in cardiac arrhythmia(prolonged the QT interval and in-creased risk of torsional ventricular tachycardia).Thus,we should pay attention to both their benefi-cial effects and potential adverse manifestation.In order to regulate the use of drugs and guide the clin-ic,this paper reviewed the electrophysiological basis and clinical characteristics of cardiac adverse re-actions of these drugs from the perspective of ion channels.

Objective:To investigate the short-term efficacy of laparoscopic total gastrec-tomy with hand-sewn esophagojejunostomy versus Roux-en-Y anastomosis.Methods:The propen-sity score matching and retrospective cohort study was conducted. The clinicopathological data of 159 patients who underwent laparoscopic total gastrectomy in Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine from October 2014 to July 2021 were collected. There were 107 males and 52 females, aged 63(range, 28?79)years. Of 159 patients, 71 cases undergoing totally laparoscopic total gastrectomy with hand-sewn esophagojejunostomy were allocated into totally laparoscopic group and 88 cases undergoing laparoscopic-assisted total gastrectomy with Roux-en-Y anastomosis were allocated into laparoscopic-assisted group, respectively. Observation indicators: (1) propensity score matching and comparison of general data of patients between the two groups after matching; (2) intraoperative and postoperative conditions; (3) perioperative complications. Propensity score matching was done by the 1:1 nearest neighbor matching method. Measurement data with normal distribution were expressed as Mean± SD, and t test was used for comparison between groups. Measurement data with skewed distribution were expressed as M(range), and the Mann-Whitney U test was used for comparison between groups. Count data were expressed as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test or Fisher's exact probability method. The rank sum test was used for comparison of ordinal data. Results:(1) Propensity score matching and comparison of general data of patients between the two groups after matching. Of 159 patients, 112 cases were successfully matched, including 56 cases in the totally laparoscopic group and 56 cases in the laparoscopic-assisted group. Before propensity score matching, age, cases with tumor located in cardia or gastric body in the totally laparoscopic group were 61(range, 30?76)years, 26, 45, respectively. The above indicators in the laparoscopic-assisted group were 65(range, 28?79)years, 50, 38, respectively. There were significant differences in the above indicators between the two groups ( Z=?2.89, χ2=6.43, P<0.05). After propensity score matching, the males and females, age, body mass index, cases of American Society of Anesthesiologists classification Ⅰ, Ⅱ, Ⅲ and Ⅳ, tumor diameter, cases with tumor located in cardia or gastric body, cases in TNM stage Ⅰ, Ⅱ and Ⅲ of patients in the totally laparoscopic group were 40, 16, (62±9)years, (22.7±2.8)kg/m 2, 22, 26, 7, 1, 3.5(range, 0.6?17.0)cm, 24, 32, 22, 9, 25. The above indicators of patients in the laparoscopic-assisted group were 38, 18, (62±10)years, (22.7±3.2)kg/m 2, 19, 32, 5, 0, 4.0(range, 0.6?15.0)cm, 23, 33, 21, 7, 28, respectively. There was no significant difference in the above indicators between the two groups ( χ2=0.17, t=?0.09, ?0.04, Z=?0.12, ?0.82, χ2=0.04, Z=?0.42, P>0.05). The elimination of age and tumor location confounding bias ensured comparability between the two groups. (2) Intraoperative and postoperative conditions: after propensity score matching, the total operation time, time of esophagojejunostomy, postopera-tive 24-hour pain numerical score and time to first out-off bed activities were (310±49)minutes, (37±10)minutes, 2.3±0.8 and (2.4±0.7)days for patients in the totally laparoscopic group, versus (344±77)minutes, (44±12)minutes, 3.1±1.2 and (2.9±1.0)days in the laparoscopic-assisted group, showing significant differences between the two groups ( t=?2.85, ?3.05, ?4.20, ?3.10, P<0.05). (3) Perioperative complications: after propensity score matching, 6 cases of the patients in the totally laparoscopic group had Clavien-Dindo grade 2 or higher complications, including 2 cases of anas-tomotic leak, 1 case of anastomotic stenosis, 1 case of pleural effusion, 1 case of abdominal infection and 1 case of intestinal obstruction. The incidence of Clavien-Dindo grade 2 or higher complications was 10.7%(6/56). In the laparoscopic-assisted group, 5 patients had Clavien-Dindo grade 2 or higher complications, including 2 cases of anastomotic leak, 1 case of abdominal infection, 1 case of intestinal obstruction and 1 case of cholangitis. The incidence of Clavien-Dindo grade 2 or higher complications was 8.9%(5/56). There was no significant difference in the incidence of Clavien-Dindo grade 2 or higher complications between the two groups ( χ2=0.10, P>0.05). Patients with anas-tomotic leak were improved after puncture and drainage, secondary surgery and conservative treat-ment, and other complications were improved after symptomatic treatment. Conclusions:Com-pared with Roux-en-Y anastomosis in laparoscopic total gastrectomy, the time of hand-sewn esophagojejunostomy and esophago-jejunal anastomosis are shorter, patients have less postopera-tive pain and faster postoperative recovery. Both methods have good peri-operative safety.

OBJECTIVE To prepare celastrol -loaded albumin nanoparticles （CLT-AN），and to investigate their activity against rheumatoid arthritis （RA）in vivo . METHODS CLT-AN was prepared by ultrasonic method . The formulation technology was optimized by single -factor test by taking particle size ，polydispersity index （PDI）and stability as indexes ，with the dosage of CLT ， the dosage of soybean oil and the ultrasonic power as factors . The physical and chemical properties of CLT -AN were investigated by transmission electron microscopy （TEM）and laser particle size analyzer ；in vitro stability and release profile were studied . A rat model of adjuvant -induced arthritis was constructed to investigate the effects of CLT -AN on joint swelling ，the levels of serum inflammatory factors [tumor necrosis factor α（TNF-α）and interleukin -1β（IL-1β）] and pathological state of joint tissue . RESULTS The optimized formulation was CLT 6.5 g，soybean oil 45 mg，ultrasonic power 490 W，ultrasonic time 8 min. CLT- AN prepared by the best formulation showed uniform and spherical morphology . Its particle size ，PDI，Zeta potential were （96.8± 1.1）nm，0.174±0.020，and（－18.6±1.7）mV，respectively. The encapsulation efficiency and drug -loading efficiency were （94.61±0.46）% and（2.42±0.21）%. There were no significant changes in particle size ，PDI，Zeta potential and encapsulation efficiency of CLT -AN within 5 days of storage at room temperature . CLT-AN was slowly released in vitro ，and the cumulative release reached 73.56% in 72 h. Compared with CLT ，CLT-AN could significantly inhibit the joint swelling of model rats ，reduced the levels of inflammatory factors TNF -α and IL -1β in serum ，and improved the pathological state of inflammatory joint tissue . CONCLUSIONS CLT-AN prepared by ultrasonic method has the appropriate particle size ，good stability ，significant sustained - release characteristics ，and excellent therapeutic efficacy against RA .

【Objective】 To investigate the effect and mechanism of achyranthes bidentata polysaccharides （ABPS） on the differentiation of bone mesenchymal stem cells （BMSCs） into nucleus pulposus-like cells. 【Methods】 Five 4-week-old SD rats were sacrificed and their BMSCs were isolated and purified by repeated adherent method. The third-generation BMSCs were treated with different concentrations of ABPS for 48 h. The cell viability was detected by MTT method. The highest concentration of ABPS was selected for subsequent experiments. According to different treatment methods， cells were divided into control group （conventional culture）， ABPS group （200 mg/L of ABPS）， Notch1 group （transfected with Notch1）， Notch1 negative control group （transfected with Notch1 negative control）， and ABPS+Notch1 group （200 mg/L of ABPS was added after transfection with Notch1）. After 14 days of induction， the cell survival rate was measured by MTT method. CollagenⅡ was detected by immunohistochemical staining. The mRNA and protein expression levels of collagenⅡ， aggrecan， Notch1 and Hes1 were detected by qPCR and Western Blotting respectively. The glucosaminoglycan （GAG） was detected by alcian blue method on the 1st， 4th， 7th， 10th and 14th day of cell culture. 【Results】 After BMSCs were treated with 400 mg/L of ABPS for 48 h， the cell survival rate was significantly lower than that in the control group （P<0.05）. When the concentration was ≤200 mg/L， ABPS had no significant toxic effect on the growth of BMSCs. After 14 days of induction， compared with the control group， A value， GAG content in the medium， the expressions of collagen II and aggrecan mRNA and protein in the ABPS group were increased， and the expressions of Notch1 and Hes1 mRNA and protein were decreased （P<0.05）. The relative cell viability， GAG content in the medium， the expressions of collagen II and aggrecan mRNA and protein in the Notch1 group were decreased， and the expressions of Notch1 and Hes1 mRNA and protein were increased （P<0.05）. ABPS+Notch1 could partially reverse the inhibitory effect of Notch1 overexpression on the differentiation of BMSCs into nucleus pulposus-like cells. 【Conclusion】 ABPS can promote the differentiation of BMSCs into nucleus pulposus-like cells， and the mechanism may be related to the inhibition of Notch1 pathway.

AIM: To investigate the effects of phorbol ester (TPA) on the anti-tumor effect and renal toxicity of cisplatin (CP). METHODS: MTT assay was used to examine the effect of TPA on the proliferation inhibition of CP in A549 and SPC-A-1 lung cancer cells. Also the effect of TPA on acute toxicity of CP was observed by once injection of high dose CP through caudal vein; The tumor-bearing mice model was explored to investigate the effect of TPA on tumor inhibition ratio and renal toxicity of CP in vivo. And the effect of TPA on renal oxidative stress induced by CP was detected. RESULTS: 1 ng/mL TPA could significantly enhance the inhibitory effect of CP on cell proliferation. In acute toxicity test, TPA could significantly reduce the toxicity of CP and prolong the survival time of animals. And the tumor weight (P<0.05), serum creatinine (P<0.05) and urea nitrogen levels (P<0.01) in TPA combined with CP group were significantly lower than those in CP group. Meanwhile, the results of HE staining showed that the renal tissue damage was significantly reduced in the combined group compared with CP group. The contents of MDA in renal tissue were decreased (P<0.01). However, the contents of GSH and the activity of SOD were increased (P<0.05) in TPA and CP combined group. CONCLUSION: TPA can enhance the inhibitory effect of CP on cell proliferation and inhibit tumor growth in tumor-bearing mice. At the same time, TPA can reduce the renal toxicity of CP, which may be related to the inhibition of renal oxidative stress induced by CP.