Objective: To develop an HPLC method for the simultaneous determination of 3 ultraviolet absorbers (benzylidene camphor sulfonic acid, camphor benzalkonium methosulfate, 4-methylbenzylidene camphor) in cosmetics.Methods: After extracted by acetonitrile-methanol-ammonium acetate aqueous solution-tetrahydrofur(30∶35∶30∶5, v/v)(for lotion and milk) or acetonitrile-methanol-ammonium acetate aqueous solution(30∶20∶50, v/v)(for cream and powder), the UV absorbers were separated on a Thermo scientific C18 column (250 mm×4.6 mm,5 μm) at 35 ℃ with methanol, acetonitrile and 0.02 mol·L-1ammonium acetate aqueous solution (pH 5.15 adjusted by glacial acetic acid) as the mobile phase with gradient elution at the flow rate of 1.0 ml·min-1, and then analyzed by a DAD detector at the wavelength of 288 nm and 300 nm.Results: Benzylidene camphor sulfonic acid, camphor benzalkonium methosulfate and 4-methylbenzylidene camphor showed good linearity within the range of 12.26-98.06 μg·ml-1(r=0.999 9), 9.31-74.48 μg·ml-1(r=0.999 9) and 6.86-54.88 μg·ml-1(r=0.999 9), respectively.The limit of detection (LOD) and the limit of quantitation (LOQ) were 4.6, 6.4 and 1.6 ng and 17.4, 15.9 and 5.5 ng, respectively.The average recovery was 99.2%(RSD=1.58%), 99.8%(RSD=2.38%) and 99.2%(RSD=2.03%)(n=36), respectively.Conclusion: The method is simple, rapid, reproducible, accurate and reliable, and can be applied in the determination of ultraviolet absorbers in cosmetics.

Objective To investigate the clinical significance of serum chemokine and hs-CRP levels in patients with type 2 diabetes mellitus with asymptomatic subclinical atherosclerosis.Methods The clinicpathological and follow-up data of 55 patients with Type 2 diabetic from 2012.1 to 2015.12 were collected and reviewed.At the same time,55 patients with non type 2 diabetes were taken as control.Determination of FPG,HbA1c,HDL-C,TC,TG,LDL-C,INS and other indicators by automatic biochemical analyzer,the application of color Doppler ultrasound equipment measurement C-IMT.Correlation using Pearson correlation analysis,risk factor analysis using multiple linear stepwise regression analysis.Results Compared with the control group,the two groups in BMI,WHR,systolic blood pressure,ankle brachial index,FPG,HbA1c,HDL-C,hsCRP,C-IMT,HOMA2-IR and serum chemokines were significantly different,with statistical significance (t =-6.31 ～5.79,P≤0.01 ～ 0.03).In 110 cases of experimental group and control group,the levels of serum chemokines were positively correlated with WHR,HOMA2-IR,C-IMT and hs-CRP (r=0.24～0.29,P=0.01～0.04).C-IMT and age,WHR,systolic blood pressure,diastolic blood pressure,FPG,HbA1c,diabetes duration,hs-CRP was positively correlated (r=0.15 ～0.68,P≤0.01～0.0.04),and the ankle brachial index was negatively correlated (r=-0.32～0.29,P≤0.01).Hs-CRP was positively correlated with HbA1c,HOMA2 IR,serum chemokine,C-IMT (r=0.25～0.32,P≤0.01～0.04),and was negatively correlated with TC and HDL-C (r=-0.27～-0.25,P all 0.02).Cox proportional hazard regression model for multivariate analysis showed that high serum chemokines,hs-CRP and HbA1c were the risk factors of C-IMT (β=0.026～0.658,SE=0.015～0.033,t=2.532～3.421,P≤0.01～0.04).Conclusion High serum levels of chemokines and hs-CRP are the risk factors of C-IMT,and it has important clinical significance for the diagnosis of asymptomatic subclinical atherosclerosis in patients with type 2 diabetes mellitus.

Objective To investigate the prognostic significance of postoperative radiotherapy (PORT) for locally advanced pulmonary adenocarcinoma with micropapillary pattern(MPPAC).Methods A total of 45 completely resected pN2-3 cases that occured from January 2012 to December 2014 at Tianjin Medical University Cancer Hospital were retrospectively analyzed.All of them were diagnosed with MPPAC by pathological diagnosis.Based on whether receiving PORT, patients were divided into radiotherapy and non-radiotherapy groups.General characteristics, overall survival and disease-free survival characteristics of the two groups were compared, respectively.Results The median overall survival (OS) of patients was 19.8 months, 1-year and 2-year overall survival rate was 79.4％ and 30.3％ , respectively.The median disease free survival (DFS) of patients was 13 months, 1-year and 2-year, and the disease free survival rate was 59.3％ and 28.9％ , respectively.The radiotherapy and non-radiotherapy groups exhibited median OS of 22.3 and 11.4 months,respectively (x2=13.329, P＜ 0.05) , and corresponding D FS of 16.2 and 10.4 months(x2 =7.972 ,P ＜0.05).The epidermal growth factor receptor gene (EGFR) mutation rate of patients was 57.14％ (20/35), In the subgroup analysis, for patients with EGFR mutation, the radiotherapy and non-radiotherapy groups showed median OS of 25.6 and 18.4 months, respectively(x2 =9.268,P ＜ 0.05) , and corresponding DFS of 21.6 and 12.6 months (P ＞ 0.05).For patients with wildtype EGFR, the radiotherapy and non-radiotherapy groups showed median OS of 21.8 and 10.6 months,respectively(x2 =9.595,P ＜ 0.05) , and corresponding DFS of 15.2 and 6.6 months(x2 =4.538,P ＜0.05).Conclusions PORT could improve survival of patients with pN2.3 MPPAC.For patients with locally advanced MPPAC after curative resection, PORT is still an integral part of treatment.

Objective To detect and investigate the expression and the effect of Prohibitin (PHB) in rats with renal interstitial fibrosis (RIF) induced by unilateral ureteral obstruction (UUO) .Methods Forty-eight Wistar male rats (6-weeks-old) were randomly assigned into 2 groups,sham-operated and model group.The model group rats were subjected to left ureteral ligation after anesthesia and the sham-operated group rats were subjected to sham operation.Six rats were killed 7,14,21,28 days after operation respectively.The renal tissues were collected.The index of RIF was calculated.The expressions of mRNA and protein of PHB were assayed by real time polymerase chain reaction and immunohistochemistry. Results Compared with sham-operation group,at each time point,the model group had significantly increased index of RIF (P < 0.01) and the obstruction for a longer period showed the higher index; the model group had significantly decreased expression of mRNA and protein of PHB (P < 0.01) and the obstruction for a longer period showed the lower expression; the model group had significantly increased expression of mRNA and protein of TGF-β1 (P < 0.01) and the obstruction for a longer period showed the higher expression.Correlation analysis showed that the index of RIF was negatively correlated with FHB (γ = -0.825) and positively correlated with TGF-β1 (γ = 0.995),while there was a positive correlation between PHB and TGF-β1 (γ = -0.786).Conclusions The lower expression of PHB in renal tissue of UUO rats might suggest that it play an important role in RIF.

Objective To explore the effects of lipoxin A4 ( LXA4 ) on lipopolysaccharide ( LPS)-induced oxidative stress in human umbilical veins endothelial cells(HUVEC) and the possible mechanism.Methods Neonatal umbilical cords were obtained from normal term pregnant women with cesarean section within 4 hours and then were used to isolate HUVEC for subculture.HUVEC were divided into four groups:control group; LPS group ( 10 μg/ml of LPS); LPS + LXA4 group ( 10 μg/ml of LPS and 100 nmol/L of LXA4); LXA4 group (100 nmol/L of LXA4) All expriments were performed after cells treated for 12 and 24 hours respectively.Immunofluorescence was used to detect the expression of Ⅷ foctor and nuclear translocation of nuclear factor-erythroid-2-related factor 2 ( Nrf2 ); the mRNA expression of Nrf2, heme oxygenase 1 (HO-1) and reduced form of nicotinamide-adenine dinucleotide quinone oxidoreductase-1(NQO1) were evaluated by reverse transcription-PCR .Results (1)The flavovirens fluorescence was observed in the cytoplasm under fluorescence microscope, which confirmed the existence of Ⅷ factor which specifically expressed in endothelial cells, especially in HUVEC.(2)Immunofluorescent results showed that in control group, Nrf2 protein expressed in the cytosol rather than in the nucleus.In LPS group, the expression of Nrf2 protein obviously increased in the nucleus while decreased in the cytosol after 12 hours.However, after LPS treatment for 24 hours, Nrf2 expression reduced in the cytosol and nucleus.In cotreatment with LPS and LXA4 group,the expression of Nrf2 protein was much higher than that in LPS group after 12 hours or 24 hours.Furthermore, Nrf2 protein also mostly expressed in the cytosol in LXA4 group.(3) After stimulation for 12 hours, compared with control group, the gene expression of Nrf2 and HO-1 were significantly enhanced in LPS group (0.581 ± 0.019 and 0.081 ±0.009, P ＜ 0.05 ) and in LPS + LXA4group(0.692 ±0.048 and 0.136 ± 0.018, P ＜ 0.05 ), the level of NQO1 mRNA in LPS group and LPS +LXA4 group were 0.381 ± 0.009 ( P ＞ 0.05 ) and 0.574 ± 0.034 ( P ＜ 0.05 ).After treatment for 24 hours,compared with control goup, the gene expressions of Nrf2 and NQO1 were down-regulated in LPS group (0.180±0.017 and 0.472 ±0.064, P＜0.05).But in LPS + LXA4 group the expression of Nrf2 and NQOI were upregulated (0.532 ± 0.051 and 0.830 ± 0.068, P ＜ 0.05, compared with treatment for LPS group).The mRNA expressions of Nrf2, HO-1 and NQO1 were increased in LPS + LXA4 group compared with LPS group ( P ＜ 0.05 ).In addition, there was no markedly difference in the expressions of Nrf2, HO1 and NQO1 between control and LXA4 group after 12 hours and 24 hours ( P ＞ 0.05 ) .Conclusion Through activating nuclear translocation of Nrf2 protein from cytoplasm, LXA4 upregulates the Nrf2downstream enzymes, such as NQO1 and HO-1 to protect HUVEC against the oxidative stress induced by LPS.

Objective To explore whether lipoxin A4 (LXA4)could prevent lipopolysaccharide (LPS)-induced human umbilical vein endothelial cells (HUVEC) monolayer hyperpermeability and its possible mechanism. Methods Human umbilical cords were obtained from women with normal pregnancy immediately after delivery from Tongji Hospital Affiliated of Tongji Medical College. Primary HUVEC were isolated from umbilical veins and subcultured, then, HUVEC were divided into four groups:control group;LPS group (10 mg/L of LPS); LPS + LXA4 group(10 mg/L of LPS and 100 nmol/L of LXA4); LPS +LXA4 + BOC-2 group [10 μmol/L of BOC-2, an effective antagonist of formyl peptide receptor like 1 (FPRL-1)]. All expriments were performed after cells were treated for 24 hours. Endothelial permeability was measured by fluorescein isothiocyan-ate labelled bovine serum albumin (FITC-BSA) clearance across the monolayer; tumor necrosis factor α(TNF-o) mRNA and secretion were detected by reverse transcriplase (RT) -PCR and ELISA assay respectively, and nuclear factor κB(NF-κB) protein change was determined by western blot. Results (1) LPS induced a significant increase in the permeability [Pa value of LPS group was (183.1 ±1.7)%], while co-administrating with LXA4 obviously attenuated this LPS-induced hyperpermeability, Pa value of LPS + LXA4 group was (103.1 ±2.2)%, LPS + LXA4 + BOC-2 group was (162.2 ± 2.8)%, control group was 100%, the permeability of HUVEC monolayer was significantly increased by LPS which was (83.1 ± 1.7)% of control (P <0.01), however, it was notably inhibited by LXA4 (P<0.05); the blockade of FPRL-1 could attenuate the effect of LXA4, that is, there was no difference between the LPS + LXA4 + BOC-2 group and the LPS group. (2) After treatment with different concentration of LPS(0,0.1, 1,10 mg/L), the mRNA expressions of TNF-α were increased (1.11 ±0.11,1.27 ± 0.03, 1.60 ± 0.06, 1.82 ± 0. 04, respectively), compared with the control group, at the concentration of 1,10 mg/L LPS, the difference was statistically significant (P<0. 05). (3) The increased levels of NF-κB and inflammatory mediator TNF-α in the LPS group were both inhibited by LXA4. Levels of NF-κB protein and TNF-o mRNA secretion in LPS treated group (0.53 ±0.06 and 0.81 ±0.09 ,respectively)were both inhibited by LXA4 (0.19 ± 0.05 and 0.41 ± 0.07, respectively, and both had significant difference, P<0.05). (4) Levels of TNF-α in HUVEC culture medium of LPS group [(31.94 ±0.01)ng/L] was significantly higher than the control group [(18.17 ± 0.03) ng/L, P<0.05], LPS + LXA4 group [(15.72 ± 0.07) ng/L] was significantly lower than the LPS group (P<0.05). Conclusion Our findings demonstrated that LXA4 could prevent the endothelial cell hyperpermeability induced by LPS in HUVEC under which the possible mechanism was through inhibiting the expression of NF-κB and its related cytokines through receptor-dependent.

OBJECTIVETo study the proliferation effect of neural stem cells (NSCs) of ginsenoside Rg1 in vitro.

METHODNSCs from the embryonic rat (E14) were isolated, 10 mg x L(-1) BrdU were added in the medium. The NSCs were incubated together with 1, 10 micromol x L(-1) ginsenoside Rg1 for 48 hours, control group were setup. Then BrdU positive cell number were detected and counted by fluorescence microscop. The Hes1 and Mash1 mRNA expression were detected by real-time RT-PCR.

RESULTThe number of BrdU positive cells in 10 micromol x L(-1) ginsenoside Rg1 group was increased significantly compared with the control group (40.5 +/- 10.9 vs 26.2 +/- 6.0, P < 0.01), and the Hes1 mRNA expression were increased significantly (1.00 +/- 0.11) vs (1.28 +/- 0.14), P < 0.05.

CONCLUSIONginsenoside Rg1 can promote the proliferation of neural stem cells in vitro, and this effect may be induced by the up-regulation Hes1 expression.

Animals ; Cell Proliferation ; drug effects ; Ginsenosides ; pharmacology ; Multipotent Stem Cells ; drug effects ; physiology ; Neurons ; drug effects ; physiology ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction

Objective:To evaluate the short-term efficacy of ustekinumab (UST) as the first-line treatment of Crohn′s disease (CD).Methods:From October 1, 2020 to March 1, 2023, at the First Affiliated Hospital of Soochow University, 64 CD patients treated with UST as first-line biologics were enrolled. The patients were classified using the Montreal classification. Clinical and endoscopic response and remission were assessed by Crohn′s disease activity index (CDAI) and simple endoscopic score for crohn′s disease (SES-CD), respectively. Clinical response was defined as a reduction in CDAI score ≥70, and clinical remission was defined as a CDAI score <150; Endoscopic response was defined as ≥50% reduction from baseline in SES-CD score, and endoscopic remission was defined as SES-CD score ≤2. The clinical response rate and clinical remission rate at week 24 and week 48, as well as the endoscopic response rate and endoscopic remission rate at week 48 were observed in CD patients (only 21 patients with endoscopic prognostic results). Mann-Whitney rank sum test was used for statistical analysis.Results:Among 64 CD patients, there were 47 males and 17 females, with an age of (33.5±13.7) years old. According to Montreal classification, there were 3 cases (4.7%) of type A1 (≤16 years old), 44 cases (68.8%) of type A2 (17 to 40 years old), and 17 cases (26.6%) of type A3 (>40 years old); 43 cases (67.2%) of type L1 (terminal ileum type), 10 cases (15.6%) of type L2 (colonic type), 8 cases (12.5%) of type L3 (ileocolonic type), 1 case (1.6%) of type L4 (upper gastrointestinal type), 2 cases (3.1%) of type L1+ L4; 23 cases (35.9%) of type B1 (non-stricturing, non-penetrating), 34 cases (53.1%) of type B2 (stricturing), 2 cases (3.1%) of type B3 (penetrating), 5 cases (7.8%) of type B2+ B3; 44 cases (68.8%) complicated with perianal lesions. Among 56 CD patients with UST maintenance therapy once every 8 weeks, the CDAI scores at week 24 and 48 after treatment were both lower than that at week 0 (64.46(30.61, 123.30), 34.24(15.77, 64.83) vs. 353.40(290.40, 391.30)), and the CDAI score at week 48 after treatment was lower than that at week 24 after treatment, and the differences were statistically significant ( Z=-9.01, -9.13, and -3.14; P<0.001, <0.001, and =0.002). The clinical response rate was 100.0% (56/56) and the clinical remission rate was 91.1% (51/56) at week 24; the clinical response rate was 100.0% (56/56) and the clinical remission rate was 98.2% (55/56) at week 48. Among 39 CD patients complicated with perianal lesions, the closure rate of anal lesions at week 24 was 87.2% (34/39) and at week 48 was 100.0% (39/39). Among 8 CD patients who received UST maintenance therapy once every 12 weeks, the CDAI scores at week 24 and 48 were both lower than that at week 0 (100.40(71.20, 171.30), 38.49(18.25, 143.50) vs. 268.00(242.60, 364.90)), and the differences were statistically significant ( Z=-3.26 and -3.36; both P<0.001). At week 24, 7 CD patients achieved clinical response and 5 CD patients achieved clinical remission. At week 48, 8 CD patients achieved clinical response and 6 CD patients achieved clinical remission. Among 5 CD patients complicated with perianal lesions, 3 CD patients achieved perianal closure at week 24 and all 5 CD patients achieved closure of perianal lesions at week 48. Among 21 CD patients who underwent endoscopic evaluation, 16 CD patients received UST maintenance therapy once every 8 weeks, the SES-CD score at week 48 was lower than that at week 0 (4.00(3.00, 7.75) vs. 9.50(7.25, 10.75)), and the difference was statistically significant ( Z=-3.43, P<0.001), among them, 9 CD patients achieved endoscopic response and 2 CD patients achieved endoscopic remission; 5 CD patients received UST maintenance therapy once every 12 weeks, there was no statistically significant difference in the SES-CD score at week 48 compared with that at week 0 (4.00(1.50, 6.50) vs. 7.00(3.50, 10.00)) ( P>0.05), among them, 3 CD patients achieved endoscopic response and 1 CD patients achieved endoscopic remission. Conclusion:UST as the first-line treatment for CD patients can achieve clinical efficacy (response or remission), and the maintenance therapy is beneficial for endoscopic remission and closure of perianal lesions.