Objective To investigate the effect on prevention and treatment autoimmune diabetes mellitus in NOD mice by oral administration porcine insulin.Methods One hundred and twenty female NOD mice were divided randomly into insulin group(n=60) and control group(n=60).Beginning from 6 week old, NOD mice were fed porcine insulin 500ul or phosphate buffered saline(PBS) 500ul seperately till 40 week old. At 6 ?12 ?20 ?25 ?30 week old,serum insulin of healthy mice was measured and pancreas were used for histology.Results Oral administration of insulin can marked decreased incidence of diabetes ( P

Objective:To observe the therapeutic effect ofcapsaicin at different concentrations on chronic knee arthritis pain model in mice.Methods:Choosing 50 healthy adult male Kunming mice builded chronic knee arthritis pain model by injecting 0.01 mL CFA (Complete Freund's Adjuvant,CFA) into left joint cavity.The model would be succeed in building after 3 weeks.The successful model mice were divided into five groups randomly (n=10):The first experimental group (saline group),the second experimental group (capsaicin excipient group),the third experimental group (0.5 ％ of capsaicin),the fourth experimental group (3 ％ of capsaicin) and the fifth experimental group (8 ％ of capsaicin).All of the mice would be observed the time of withdrawal latencies from the thermal heated surface after administration of one,four and seven hours,and thermal withdrawal time within 60 days after the injection.Results:①The physiological saline group compared with excipient group,the thermal withdrawal time had no statistically significant difference (P＞0.05)after administration of one,four and seven hours,and thermal withdrawal time within 60 days.②The acute pain duration of the third group would disappear after capsaicin injection 7 hours,four hours for the fourth group,and one hour for the fifth group.③The duration of analgesia of the third group,lasted for 18.9± 1.1 days;The analgesia time of the fourth group lasted for 33.7± 1.0 days;The analgesia time of the fifth group lasted for 58.2± 1.2 days.Conclusions:Capsaicin has analgesic effects on chronic knee pain model in mice induced by CFA,and the days of analgesia increases with the concentration of capsaicin.

OBJECTIVE To explore the effect of clofenotane (DDT) on epithelial-mesenchymal transition (EMT) and the relevant molecular mechanism in human colorectal cancer cells. METHODS Human colorectal cancer cells DLD1 were treated with DDT 0.01, 0.1, 1.0, 10.0 and 100.0 nmol·L-1 for 48 h. Then, the morphology of DLD1 cells was observed. mRNA levels of E-cadherin, N-cadherin, vimentin and Snail1 were detected by real-time PCR. Protein expression of STAT3 signaling pathway of proteins STAT3 and p-STAT3 was detected by Western blotting. STAT3 inhibitor WP1006 (5μmol · L-1) was added to determine its impact on DDT-induced alternation of STAT3/Snail1 signaling and EMT-related molecules. Protein expression of STAT3 and p-STAT3 was detected by Western blotting and mRNA levels of E-cadherin, N-cadherin, Vimentin and Snail1 were detected by real-time PCR. RESULTS DLD1 cell morphology was changed after exposure to DDT 0.01-100.0 nmol · L- 1. Meanwhile, real-time PCR showed that the mRNA level of E-cadherin was significantly decreased compared with normal cell control (P<0.01), which was 42.4±2.8%of that in the normal control group. The mRNA levels of N-cadherin, Vimentin and Snail1 were significantly increased (P<0.01), which were 1.91±0.1, 1.5±0.2 and 1.5±0.1 times that of the normal control group. DDT 0.1, 1.0 and 10.0 nmol · L-1 exposure induced up-regulation of STAT3 and p-STAT3 protein levels (P<0.01), which were 2.1 and 1.8 times that of the normal control group. The addition of STAT3 inhibitor WP1066 (5 μmol · L-1) prevented STAT3 from phosphorylation as well as the up-regulation of Snail1(P<0.01), which was (56.3 ± 0.9)% that of the DDT 1.0 nmol · L-1 treat?ment group. Compared with DDT treatment alone, the mRNA levels of EMT-related molecules were remarkably reversed by WP1066 (5 μmol · L- 1) co-treatment, increasing E-cadherin but decreasing N-cadherin and vimentin in DLD1 cells(P<0.01), which were 50.2±2.9%and 61.6±6.1%of those in the DDT 1.0 nmol · L- 1 treatment group, respectively. CONCLUSION DDT alters the expressions of EMT-related molecules including E-cadherin, N-cadherin and vimentin via STAT3/Snail1 signaling, thus promoting the EMT process in human colorectal cancer cells. This progress may be closely related to DDT-induced colorectal cancer development.

[Objective]To establish a novel noninvasive fluorescent animal model for endometriosis in vitro and in vivo.[Methods]Adenovirus encoding enhancing green fluorescent protein(Ad-eGFP)was used to transfect endometrial glandular cells and stromal cells(cells transfection and injection,Method No.1),and fragments(tissues transfection and injection,Method No.2).Transfection efficiencies were compared between the two methods in vitro.Then GFP transfected glandular cells and stromal cells suspension were injected into nude mice subcutaneously(Method No.1),taking Method No.2 as a comparison.In vivo observation last for 25 days,and positive rates and duration times of fluorescent lesions were calculated.Histological examination was used to confirmed lesion formation.[Results]On the fifth day after injection,lesion positive rate of Method No.1 was 88.9%,which was statistically significantly higher than that of Method No.2(22.2%),P=0.015＜0.05.The fluorescent positive duration of Method No.1 and No.2 were 12 ± 8 days and 7±4 days.The structures of lesions were all identified as human original endometrium by histological examination,including HE staining and immunofluoresceney.[Conclusion]Noninvasive animal model of endometriosis can be built up by subcutaneously injection of Ad-EGFP transfected endometrial glandular cells and stromal cells suspension with higher positive rate and longer observation time

[ ABSTRACT] AIM:To investigate the effects of microRNA-378*( miR-378*) on the survival and apoptosis of human mesenchymal stem cells ( hMSCs ) .METHODS: The expression of miR-378* was determined by microRNA arrays and quantitative real-time PCR ( qRT-PCR) .H2 O2 was used to induce hMSCs apoptosis.By transfection of miR-378*mimic or inhibitor, we up-regulated or down-regulated miR-378* expression in hMSCs.The effect of miR-378*and connective tissue growth factor ( CTGF) on hMSC survival and apoptosis were detected by MTT, LDH, caspase-3/7 and TUNEL assays.RESULTS:The expression of miR-378*was up-regulated in the old hMSCs compared with the young hMSCs.H2 O2 increased the expression of miR-378*, decreased the expression of CTGF.Up-regulation of miR-378*re-sulted in increasing apoptosis and decreasing survival of hMSCs.Conversely, down-regulation of miR-378*resulted in de-creasing cell apoptosis and increasing survival.The regulation of miR-378*on hMSC apoptosis and survival was attenuated by inhibiting the expression of miR-378* and CTGF together.Direct repression of CTGF expression inhibited the hMSC survival and increased apoptosis.CONCLUSION:miR-378*enhances apoptosis of hMSCs by repressing the expression of CTGF.

AIM: To investigate the effect of microRNA-708-5p (miR-708-5p) on the migration of human mesenchymal stem cells (hMSCs).METHODS:The expression of miR-708-5p was determined by miRNA arrays and re-al-time PCR.By transfection of miR-708-5p mimic or inhibitor, the up-regulation or down-regulation of miR-708-5p ex-pression in hMSCs was evaluated .The cell scratch and Transwell tests were used to detect the migration capability of hM-SCs.The effects of transmembrane protein 88 (TMEM88), a miR-708-5p target gene, onβ-catenin expression and migra-tion of hMSCs were detected .RESULTS:The expression of miR-708-5p was down-regulated in the old hMSCs compared with the young hMSCs.Up-regulation of miR-708-5p resulted in increasing migration of hMSCs.Conversely, down-regula-tion of miR-708-5p resulted in decreasing cell migration .The expression of TMEM88 was up-regulated in the old hMSCs compared with the young hMSCs , while the expression of β-catenin was down-regulated.Directly repression of TMEM88 expression increased the β-catenin expression and migration of hMSCs .The regulation of miR-708-5p on hMSCs was atten-uated by inhibiting the expression of miR-708-5p and TMEM88 together.CONCLUSION:miR-708-5p increases β-cate-nin expression and Wnt/β-catenin activity by repressing TMEM 88, thus enhancing the migration of hMSCs .

Objective To study the angle and articular facet curvature of lumbar zygapophyseal joints in adults.Methods The lumbar zygapophyseal joints in 120 healthy subjects without lumbar diseases were detected using spiral CT and mutiple planner re-construction.The angle and articular facet curvature of zygapophyseal joints were measured.The differences in the measured param-eters between male and female or different age groups were compared.Results ①There were no significant differences in the angle of the same lumbar zygapophyseal joints between males and females or different age groups (P >0.05);however,the significant differences in the angle between different lumbar segments were found (P <0.05).②There were no significant differences in articu-lar facet curvature of the same lumbar zygapophyseal joints between males and females or different age groups (P >0.05);however, there were significant differences in the curvature between different lumbar segments (P <0.05).The maximum articular facet cur-vature of L3-L4 was 22.1°±6.0°.Conclusion The angle and articular facet curvature of lumbar zygapophyseal joints varies from different lumbar segments with different contributions for the lumbar stability.

AIM:To investigate the effects of microRNA-486-5p (miR-486-5p) on the senescence of human mesenchymal stem cells ( hMSCs).METHODS: The expression of miR-486-5p was determined by miRNA arrays and real-time PCR.By transfection of miR-486-5p mimic or inhibitor, up-regulation or down-regulation of miR-486-5p expres-sion in hMSCs was established.The effect of miR-486-5p and silence information regulator 1 (SIRT1) on hMSC telomerase activity and senescence were detected byβ-galactosidase staining.RESULTS:The expression of miR-486-5p was up-regu-lated in the old hMSCs compared with the young hMSCs.Up-regulation of miR-486-5p resulted in increasing senescence of hMSCs.Conversely, down-regulation of miR-486-5p resulted in decreasing cell senescence.The expression of SIRT1 and telomerase reverse transcriptase ( TERT) was down-regulated in the old hMSCs compared with the young hMSCs.Directly repression of SIRT1 expression inhibited the hMSC TERT protein expression and telomerase activity, but increased cell se-nescence.The regulation of miR-486-5p on hMSC senescence was attenuated by inhibiting the expression of miR-486-5p and SIRT1 together.CONCLUSION:miR-486-5p enhances senescence of hMSCs by decreasing the expression of SIRT1 and telomerase activity.

Objective:To investigate the effects of femtosecond laser assistcataract extraction on the levels of inflammatory factors in aqueous humor, corneal perception and high order aberration.Methods:Forty-eight patients (56 eyes) undergoing cataract surgery in the Shouguang People′s Hospital from June 2018 to February 2020 were enrolled in this study, and they were divided into observation group (24 patients and 27 eyes, femtosecond laser cataract phacoemulsification) and control group (24 patients and 29 eyes, coaxial microincision phacoemulsification) by random number table method. The levels of prostglndin E2 (PGE2), interleukin (IL)-6 and IL-1β in aqueous humor were detected before and after operation. Upper corneal and central corneal perception thresholds were recorded before surgery, 7 d after surgery, 1 month after surgery and 3 months after surgery. High order aberrations of the whole eye and cornea at 4mm pupil diameter before and after operation were recorded.Results:After operation, the levels of PGE2, IL-6 and IL-1β in aqueous humor in the observation group were lower than those in the control group: (45.62 ± 5.71) ng/L vs. (63.65 ± 5.62) ng/L, (15.25 ± 3.21) ng/L vs. (24.59 ± 6.13) ng/L, (17.16 ± 1.28) ng/L vs. (25.74 ± 4.62) ng/L, the differences were statistically significant ( P<0.05). The upper and central corneal perception threshold of observation group after operation for 7 d were higher than those in the control group: 4.55 ± 0.50 vs. 4.20 ± 0.33, 5.68 ± 0.49 vs. 5.52 ± 0.45, the differences were statistically significant ( P<0.05). The indexes of high order aberration of the whole eye decreased in the two groups ( P<0.05), while the indexes of cornea did not change significantly. The indexes of high order aberration of the whole eye and cornea between before and after surgery between the two groups had no significant differences ( P>0.05). Conclusions:Femtosecond laser assist cataractextraction can relieve inflammation, reduce the whole eye high order aberration, but not affect the cornea, and can improve corneal perception.

Objective: To conduct a systematic review and meta-analysis of studies evaluating the oncological and fertility outcomes of early-stage endometrial cancer (EC) treated with the levonorgestrel-releasing intrauterine system (LIUS)-based regimens. Methods: The Meta-analyses Of Observational Studies in Epidemiology statement for meta-analyses was followed. Searches were conducted on MEDLINE, Embase, PubMed, Preprints, and the Cochrane Central Register of Controlled Trials from January 1990 to August 4, 2022. The Joanna Briggs Institute Critical Appraisal Checklist was used for quality assessment. The primary endpoint was the complete response (CR) rate and the secondary endpoints were relapse, pregnancy, and live birth rate. Results: A total of 25 studies (821 women) were included. The CR rate of LIUS-based regimens was 63.4% (95% confidence interval [CI]=52.3%–73.2%), with 29.6% (95% CI=23.3%–36.8%) of cases experiencing recurrence during follow-up. In sensitivity analyses, patients younger than 45 years of age with a body mass index <30 kg/m2 who were treated with LIUS-based regimens achieved a high CR rate of 84.6% (95% CI=80.3%–88.1%) over a median follow-up of more than 24 months. Overall pregnancy and live birth rates were 37.9% (95% CI=24.1%–53.9%) and 39.3% (95% CI=24.0%–57.0%), respectively. No statistical differences were apparent in CR or relapse rates among the LIUS+GnRH agonist, LIUS+oral progesterone, or hysteroscopic resection followed by LIUS subgroups. Conclusion LIUS-based therapies are viable for the conservative management of early-stage endometrioid EC on CR and fertility outcome.