BACKGROUND: Puerarin, the main effective component of Chinese herb, Radix puerariae, is isoflavone monomer, which can counteract learning and memory impairment induced by scopolamine or D-galactose etc.OBJECTIVE: To investigate the protective effects of puerarin on β-amYloid peptide-induced learning and memory impairment of model mouse of dementia and the changes of superoxide dismutase activity and malondialdhehyde content in brain and blood.DESIGN: Randomized controlled trailSETTING: Department of Pharmacology, Capital University of MedicalSciencesMATERIALS: The experiment was conducted in Departmentof Pharmacology of Capital University of Medical Sciences from March to June 2002.A total of 40 ICR mice were selected and randomly divided into 4 groups:pseudooperation group, dementia model group, puerarin 25 mg/kg group and puerarin 50 mg/kg group, with 10 in each group.METHODS: ①Model preparation: After anaesthesia with pentobarbital sodium, single intraventricular injection of 3 μL β-amyloid peptide was conducted from right side on each mouse in dementia model group, puerarin 25 mg/kg group and puerarin 50 mg/kg group under aseptic manipulation. The same operation was carried out on the mouse in pseudooperation group but without injection of β-amyloid peptide. ②Giving medicine:10 mL/kg physiological saline was intraperitoneally injected into the mouse in pseudooperation group and model group; 25 mL/kg puerarin was intraperitoneally injected to the mouse in 25 mg/kg puerarin group; 50 mL/kg puerarin was intraperitoneally injected to the mouse in 50 mg/kg puerarin group.The medicines were given to each group from the day of model preparation on and behavioral test was carried out 12 days later. ③ Morris water maze examination was used to detect learning and memory ability of the mice.Time for finding the platform (escape latency) in 2 minutes, swimming distance, original angle and search strategy were recorded as learning results.④When the above experiment was finished, anaesthesia with ether was applied to the mice and blood was collected from the orbit to prepare serum.After that, the mice were put to death by decapitation and the tissue of right-brain of the mice were rapidly took out to prepare cerebral homogenate in ice bath, then superoxide dismutase activity and malondialdhehyde content were determined in brain and serum.MAIN OUTCOME MEASURES: ①Escape latency, swimming distance,search strategy and original angle for the mouse in each group to reach the latform. ②Superoxide dismutase activity and malondialdhehyde content in brain and blood of the mouse in each group.RESULTS: All the 40 mice were involved in result analysis. ① Escape latency and swimming distance were shortened in puerarin 25 mg/kg and 50 mg/kg groups (P ＜ 0.05-0.01). The results of search strategy and original angle indicated that as the number of training days increased, the frequency of randomization+magin strategy gradually decreased; The decreasing rates and extents in pseudooperation group and puerarin 25 mg/kg and 50 mg/kg puerarin were more rapid than that in dementia model group,meanwhile, the increasing rates and extents of frequency of tendency+directness strategy in these groups were more rapid than that in dementia model group. There was no significant difference in original angle between groups (P ＞ 0.05). ② The content of superoxide dismutase increased and that of malondialdhehyde decreased in brain and blood of the model mouse in puerarin 25 mg/kg group andpuerarin 50mg/kg group (P ＜ 0.05 -0.01).CONCLUSION: Puerarin can counteract the neurotoxicity of β-amyloid peptide, which improves learning and memory of model mouse. It is not relevant to the dosage but probably related with elimination of cerebral free radical and improvement of antioxidation activity.

Objective To set up a technical platform of reverse genetics based on the 8 plasmid.virus rescue system of cold-adapted influenza virus strain. Methods The cold-adapted, temperature sensitive, live attenuated influenza virus strain A/AnnArbor/6/60(H2N2)was chosen as the master donor virus(MDV)for rescue research,and its six internal gene fragments PB2,PB1,PA,NP,M and NS were artificially synthesized. Meanwhile, five amino acid mutations have been introduced as tags. Six fragments were ligated with modified pAD3000 for the construction of rescue plasmid. Six transcription/expression plasmids(pMDV-A-PB2,pMDV-A-PB1,pMDV-A-PA,pMDV-A-NP,pMDV-A-M,and pMDV-A-NS)were obtained, and their sequences were accurate. Results The reassorted virus named as rMDV-A contains HA and NA gene segments derived from PR8 strain along with six gene segments,PB2,PB1,PA,NP,M and NS,from MDV. The COS-1 cells were co-transfected with eight recombinant plasmids. The results showed that a cold-adapted, attenuated reassortant influenza A virus with hemagglutination activity was rescued successfullv bv"6+2" combination of MDV and PR8, and the allanotoic fluid of the injected eggs gave a posigenes of A/AA/6/60 used as backbone has provided experimental materials for further research on the gene function and novel vaccine candidate of cold-adapted, attenuated human influenza virus.

Objective:To observe the effects of Sanjia Powder on myocardial fibrosis in diabetic cardiomyopathy (DCM) model rats; To elaborate its mechanism.Methods:A DCM rat model was constructed using STZ single intraperitoneal injection combined with high-sugar and high-fat feeding. The model rats were divided into model group, TCM group, Western medicine group, miR-21 inhibition group, and miR-21 inhibition control group using a random number table method, with 10 rats in each group; another 10 SD rats were set as normal group. TCM group was orally administered with Sanjia Powder water decoction at a dose of 6.2 g/kg, and the Western medicine group was administered metformin 250 mg/kg + enalapril 45 mg/kg by gavage. miR-21 inhibition group, miR-21 inhibition control group, normal group, and model group were given the same volume of distilled water by gavage, once a day, for 4 weeks; at the same time, the normal group, the model group, TCM group, and the Western medicine group were injected with the same amount of normal saline in the tail vein. The miR-21 inhibition group received tail vein injection of miR-21 inhibitor at a dose of 20 mg/kg, and the miR-21 inhibition control group received tail vein injection of miR-21 inhibitor NC at a dose of 20 mg/kg, twice a week, for 4 weeks. The morphological changes of myocardial tissue were observed using HE; the expression of Collagen-Ⅰ in myocardial tissue was detected using Western Blot; the expressions of endothelial cell marker CD31 and stromal cell marker FSP1 in myocardial tissue were detected using immunofluorescence method.Results:Compared with the model group, the pathological morphology of myocardial tissue improved in the TCM group, Western medicine group, and miR-21 inhibition group, while the expressions of Collagen-Ⅰ and FSP1 decreased ( P<0.01) and the expression of CD31 increased ( P<0.01); there was no statistically significant difference in the expressions of Collagen-Ⅰ, CD31, and FSP1 between the TCM group and the miR-21 inhibition group ( P>0.05). Conclusion:Sanjia Powder can improve myocardial fibrosis in rats with diabetic cardiomyopathy, and its mechanism may be related to the up-regulation of CD31 expression and down-regulation of FSP1 expression, thereby inhibiting endothelial mesenchymal transition, and may also be related to the inhibition of miR-21 expression.

Objective To investigate the effects of CD226 knockout ( KO) on obese mice fed with high fat diet and to analyze the composition of immune cells in CD226KO obese mice for further elucidating the immunological mechanism of CD226 involved in high fat diet-induced obesity. Methods Both wild-type ( WT) and CD226KO mice were randomly divided into two groups, high-fat and normal diet groups, and fed for 14 weeks to establish the type 2 diabetes model. Immune cells in mouse spleen and peripheral blood were analyzed by flow cytometry. In in vitro experiments, NK92-MI cells were infected with pshRNA-CD226 lenti-virus to silence CD226 expression, and then qPCR was performed to detect the expression of Foxp3, TNF-αand IFN-γ at mRNA level. Results In the high-fat diet groups, CD226KO mice had lower blood glucose, serum insulin and HOMA-IR than WT mice, but higher HOMA-IS and HOMA-β. CD226KO could reduce compensatory hyperplasia of islet tissue, and significantly down-regulate the proportion of spleen NK cells in mice. The proportion of CD3-CD49b+CD25+Foxp3+regulatory NK cells (NKreg) increased significantly in CD226KO mice. CD226KO could significantly increase Foxp3 expression in NK92-MI cells and decrease the expression of TNF-α and IFN-γ. Conclusions CD226KO can alleviate insulin resistance, increase the number of islet β-cell and improve islet β-cell function in obese mice. The mechanism might be related to the up-regulation of Foxp3+ NKreg ratio.

OBJECTIVE: To explore the correlation between altered levels of neurotransmitters in the frontal lobe and hippocampus and behavioral abnormalities in a Clock variant mice modeling bipolar disorder manic disorder. METHODS: Open field test and Elevated plus-maze test were carried out on the Clock mutant and wild-type control groups. The frontal lobe and hippocampus of Clock mutant mice and controls were dissected, and neurotransmitters in tissue extracts were analyzed by high-performance liquid chromatography and mass spectrometry. The concentration of neurotransmitters and behavioral indicators were assessed by t test and Pearson correlation analysis using SPSS 22.0. RESULTS: The Clock mutant mice showed a significant increase in activity, albeit with no difference in the level of anxiety from the wild-type controls, which suggested that the Clock mutant mice can be used as a model for manic attack of bipolar disorder. Altered neurotransmitter levels were detected in the frontal and hippocampal regions, including elevated histamine in the left hippocampus, reduced histamine in the right hippocampus, reduced gamma-aminobutyric acid (GABA) in bilateral hippocampus, elevated dihydroxyphenylalanine (DOPA) in the left frontal lobe and reduced DOPA in the right hippocampus, and decreased glutamine in bilateral frontal lobes. The reduced glutamine in the left frontal lobe and GABA in the right hippocampus correlated with the increased activity of Clock mutant mice. CONCLUSION Clock mutant mice showed abnormal behavior with increased activity. Reduced glutamine in the left frontal lobe and GABA in the right hippocampus were correlated with increased activity.

Objective:To investigate the influencing factors of textbook outcomes in liver surgery (TOLS) after radical resection of gallbladder carcinoma.Methods:The retrospective case-control study was conducted. The clinicopathological data of 530 patients who underwent radical resection of gallbladder carcinoma in 15 medical centers, including the First Affiliated Hospital of Army Medical University et al, from January 2014 to January 2020 were collected. There were 209 males and 321 females, aged (61±10)years. Patients underwent radical resection of gallbladder carcinoma, including cholecystectomy, hepatectomy, invasive bile duct resection, and lymph node dissection. Observation indicators: (1) situations of TOLS; (2) influencing factors of TOLS. Measure-ment data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data between groups was conducted using the Mann-Whitney U test. The univariate analysis was conducted using the corresponding statistical methods based on data type, and variables with P<0.10 were included in multivariate analysis. Multivariate analysis was conducted using the Logistic stepwise regression model. Results:(1) Situations of TOLS. All 530 patients underwent radical resection of gallbladder carcinoma, and there were 498 cases achieving R 0 resection, 508 cases without ≥grade 2 intra-operative adverse events, 456 cases without postoperative grade B and grade C biliary leakage, 513 cases without postoperative grade B and grade C liver failure, 395 cases without severe com-plications within postoperative 90 days, 501 cases did not being re-admission caused by severe com-plications within postoperative 90 days. Of the 530 patients, 54.53%(289/530) of patients achieved postoperative TOLS, while 45.47%(241/530) of patients did not achieve postoperative TOLS. (2) Influencing factors of TOLS. Results of multivariate analysis showed that American Society of Anesthesiologists classification >grade Ⅱ, preoperative jaundice, T staging as T3?T4 stage, N staging as N2 stage, liver resection as right hemi-hepatectomy, and neoadjuvant therapy were independent factors influencing TOLS in patients undergoing radical resection of gallbladder carcinoma ( odds ratio=2.65, 1.87, 5.67, 5.65, 2.55, 3.34, 95% confidence interval as 1.22?5.72, 1.18?2.95, 2.51?12.82, 2.83?11.27, 1.41?4.63, 1.88?5.92, P<0.05). Conclusion:American Society of Anesthesiologists classification >grade Ⅱ, preoperative jaundice, T staging as T3?T4 stage, N staging as N2 stage, liver resection as right hemi-hepatectomy, and neoadjuvant therapy are independent factors influencing TOLS in patients undergoing radical resection of gallbladder carcinoma.

Objective:To investigate the influence of curative-intent resection with textbook outcomes of liver surgery (TOLS) on long-term prognosis of gallbladder carcinoma (GBC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 824 patients with GBC in the national multicenter database of Biliary Surgery Group of Elite Group of Chinese Journal of Digestive Surgery, who were admitted to 15 medical centers from January 2014 to January 2021, were collected. There were 285 males and 539 females, aged (62±11)years. According to the evalua-tion criteria of TOLS, patients were divided into those who achieved TOLS and those who did not achieve TOLS. Measurement data with normal distribution were represented as Mean± SD, and com-parison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data were conduc-ted using the Mann-Whitney U test. The Kaplan-Meier method was used to calculate survival rate and draw survival curve, and the Log-rank test was used for survival analysis. The COX stepwise regression model with backward Wald method was used for univariate and multivariate analyses. Results:(1) Achievement of TOLS. Of the 824 patients undergoing curative-intent resection for GBC, there were 510 cases achieving TOLS and 314 cases not achieving TOLS. (2) Follow-up. Of the 824 patients undergoing curative-intent resection for GBC, after excluding 112 deaths within 90 days after discharge, 712 cases were included for the survival analysis. The median follow-up time, median overall survival time and 5-year overall survival rate of the 510 patients achieving TOLS were 22.1(11.4,30.1)months, 47.6(30.6,64.6)months and 47.5%. The median follow-up time, median overall survival time and 5-year overall survival rate of the 202 patients not achieving TOLS were 14.0(6.8,25.5)months, 24.3(20.0,28.6)months and 21.0%. There was a significant difference in overall survival between patients achieving TOLS and patients not achieving TOLS ( χ2=58.491, P<0.05). (3) Analysis of factors influencing prognosis of patients. Results of multivariate analysis showed that TOLS, carcinoembryonic antigen (CEA), CA19-9, poorly differentiation of tumor, T2 stage of eighth edition of American Joint Committee on Cancer (AJCC) staging, T3 and T4 stage of eighth edition of AJCC staging, N1 stage of the eighth edition of AJCC staging, N2 stage of the eighth edition of AJCC staging, adjuvant therapy were independent factors influencing overall survival time of patients undergoing curative-intent resection for GBC ( hazard ratio=0.452, 1.479, 1.373, 1.612, 1.455, 1.481, 1.835, 1.978, 0.538, 95% c onfidence interval as 0.352-0.581, 1.141-1.964, 1.052-1.791, 1.259-2.063, 1.102-1.920, 1.022-2.147, 1.380-2.441, 1.342-2.915, 0.382-0.758, P<0.05). Conclusion:Patients under-going curative-intent resection for GBC with TOLS can achieve better long-term prognosis.

Rheumatoid arthritis is a chronic, systemic autoimmune disease predominantly based on joint lesions with an extremely high disability and deformity rate. Several drugs have been used for the treatment of rheumatoid arthritis, but their use is limited by suboptimal bioavailability, serious adverse effects, and nonnegligible first-pass effects. In contrast, transdermal drug delivery systems (TDDSs) can avoid these drawbacks and improve patient compliance, making them a promising option for the treatment of rheumatoid arthritis (RA). Of course, TDDSs also face unique challenges, as the physiological barrier of the skin makes drug delivery somewhat limited. To overcome this barrier and maximize drug delivery efficiency, TDDSs have evolved in terms of the principle of transdermal facilitation and transdermal facilitation technology, and different generations of TDDSs have been derived, which have significantly improved transdermal efficiency and even achieved individualized controlled drug delivery. In this review, we summarize the different generations of transdermal drug delivery systems, the corresponding transdermal strategies, and their applications in the treatment of RA.