[Clinical outcome of combined over-the-top reconstruction with modified Lemaire procedure in one-stage revision anterior cruciate ligament reconstruction].

Objective: To investigate the clinical effect of over-the-top (OTT) reconstruction of the anterior cruciate ligament (ACL) combined with the modified Lemaire technique in one-stage ACL revision. Methods: It's a retrospective study. The clinical data of 37 patients who underwent one-stage revision of ACL using OTT combined with modified Lemaire technique from April 2020 to May 2023 in the Department of Sports Medicine of Xinhua Hospital Affiliated to Dalian University were retrospectively analyzed. There were 28 males and 9 females with a mean age of (32.7±5.7) years. The postoperative knee function and surgical revision effect were evaluated by subjective and objective evaluation indexes of the knee joint before and 3 months and 24 months after surgery. The subjective evaluation indexes included the International Knee Documentation Committee (IKDC) score, Lysholm score, and Tegner score; and the objective evaluation indexes included the anterior-posterior static displacement distance of the tibia, tibia-femur relative rotation angle, the Lachman test, the axial shift test, and the ratio of the loss of muscle strength of the lower limb. Results: At the final follow-up, all patients returned to pre-injury motor level, with significant improvements in IKDC scores (45.3±6.8 preoperatively, 67.5±4.7 and 93.2±2.3 at 3 months and 2 years after the operation, respectively), Lysholm scores (57.2±2.6 preoperatively, 72.6±2.9 and 89.7±3.7 at 3 months and 2 years after the operation, respectively), and Tegner scores (3.1±0.7 preoperatively, 4.9±0.6 and 5.8±1.3 at 3 months and 2 years after the operation, respectively) (all P<0.001). The anterior-posterior static displacement distance of the tibia reduced significantly [from (5.2±0.5) mm before the operation to (1.4±0.5) mm at 2 years postoperatively) (P<0.001)]; and the relative rotation angle of the tibia-femur restored to the normal physiological range, it was reduced from 6.2°±1.2° before the operation to 1.7°±0.3° 2 years after (P<0.001). The Lachman test and axial shift test at 2 years postoperatively were all negative. The muscle strength loss ratio of the affected lower limb was significantly better than that before the operation (P<0.001). Conclusion: The use of OTT reconstruction combined with modified Lemaire technique for revision of ACL in patients with ACL reconstruction failure can circumvent the tunnel problem for one-stage revision and better restore the stability and function of the knee joint, with ideal clinical results.

[Risk factors analysis and the development and validation of a clinical prediction model for acute mesenteric ischemia-induced intestinal infarction].

Objective: To investigate the risk factors of intestinal infarction caused by acute mesenteric ischemia (AMI), and develop and validate a clinical prediction model of Nomogram plot for intestinal infarction caused by AMI. Methods: From January 2014 to December 2020, 201 patients with AMI in the Affiliated Hospital of Yunnan University were selected as the modeling group, and 105 patients with AMI in the First Affiliated Hospital of Kunming Medical University were collected as the validation group. The age of patients in the modeling group was (62±12) years, and there were 70 males and 131 females. The age of patients in the validation group was (62±10) years old, and there were 69 males and 36 females. The clinical data including clinical manifestations, laboratory indicators and imaging indicators of patients in the two groups were retrospectively analyzed, and univariate and multivariate binary logistic regression analyses were performed to screen out risk factors for the progression of AMI to intestinal infarction. A Nomogram plot clinical prediction model for AMI causing intestinal infarction was developed using R software, and the differentiation, calibration and net clinical benefit of this Nomogram plot were evaluated by C-index, calibration curve and clinical decision analysis curve. Results: Shock (OR=13.69, 95%CI: 2.31-88.87), body temperature ≥38.0 ℃ (OR=6.39, 95%CI: 1.85-22.11), white blood cells (WBC) ≥18×109/L (OR=1.19, 95%CI: 1.03-1.37), intestinal changes on CT (OR=0.17, 95%CI: 0.04-0.66), peritoneal irritation (OR=0.06, 95%CI: 0.01-0.26), and pH≤7.34 (OR=0.00, 95%CI: 0.00-0.01) were risk factors for intestinal infarction caused by AMI. Body temperature ≥38.0 ℃ (sensitivity: 81.5%, specificity: 87.0%), WBC ≥18×109/L (sensitivity: 84.0%, specificity: 83.0%) and pH ≤7.34 (sensitivity: 70.4%, specificity: 76.7%) were cut-off values for progression to intestinal infarction in patients with AMI. The Nomogram plot prediction model of intestinal infarction induced by AMI was established by using the above parameters and validated internally and externally. The C index of validation group and modeling group were 0.96 (95%CI: 0.921-0.999) and 0.98 (95%CI: 0.962-0.996), respectively. The calibration curves of the validation group and the modeling group were shaped near the reference line, and the deviation from the reference line was low. The clinical decision analysis curves of validation group and modeling group showed that the clinical prediction model of Nomogram could bring better clinical net benefit for AMI patients. Conclusions: Shock, temperature, white blood cells, intestinal changes on CT, peritoneal irritation sign, and PH are independent risk factors for AMI-induced intestinal infarction. The Nomogram plot established by these factors can effectively predict the probability of AMI patients progressing to intestinal infarction.

Near-GeV Electron Beams at a Few Per-Mille Level from a Laser Wakefield Accelerator via Density-Tailored Plasma.

A simple, efficient scheme was developed to obtain near-gigaelectronvolt electron beams with energy spreads of few per-mille level in a single-stage laser wakefield accelerator. Longitudinal plasma density was tailored to control relativistic laser-beam evolution, resulting in injection, dechirping, and a quasi-phase-stable acceleration. With this scheme, electron beams with peak energies of 780-840 MeV, rms energy spreads of 2.4‰-4.1‰, charges of 8.5-23.6 pC, and rms divergences of 0.1-0.4 mrad were experimentally obtained. Quasi-three-dimensional particle-in-cell simulations agreed well with the experimental results. The dechirping strength was estimated to reach up to 11 TeV/mm/m, which is higher than previously obtained results. Such high-quality electron beams will boost the development of compact intense coherent radiation sources and x-ray free-electron lasers.

[The prognostic value of myocardial infarct size measured by cardiovascular magnetic resonance in patients with acute ST-segment elevation myocardial infarction].

Objective: To investigate the prognostic value of infarct size (IS) in patients with acute ST-segment elevation myocardial infarction (STEMI) underwent primary percutaneous coronary intervention (PCI). Methods: A total of 104 patients with acute STEMI who underwent primary PCI treatment in Shengjing Hospital of China Medical University from February 2017 to November 2018 were included in the present study. All patients underwent cardiovascular magnetic resonance (CMR) within one week after primary PCI treatment. The subjects were followed up for two years. Major adverse cardiac events (MACE) included new onset congestive heart failure and/or recurrent nonfatal myocardial infarction and/orcardiac death. The optimal IS cutoff value for MACE was determined by receiver operating character (ROC) curve. Based on the IS cutoff value, the patients were divided into the high IS group and the low IS group. Clinical characteristics between the two groups were compared. A cox regression model was used to analyze the prognostic value of IS in acute STEMI patients treated with primary PCI for the adverse events. Results: The IS cutoff value determined by ROC curve was 13.55%. 50 patients were in the high IS group (IS≥13.55%) and 54 patients were in the low IS group (IS<13.55%). More female patients [14 cases (28.0%) vs. 6 cases (11.1%)] were in the IS group, and a higher proportion of patients in the high IS group had anterior myocardial infarction [27 cases (54.0%) vs. 16 cases (29.6%)] or microvascular obstruction [32 cases (64.0%) vs. 16 cases (29.6%)]. White blood cell counts [11.25(8.90, 13.38) ×109/L vs. 9.25(7.58, 11.00) ×109/L], troponin I levels [50.63(16.56, 76.30)µg/L vs. 16.58(2.66, 38.42)µg/L] and brain natriuretic peptide levels [178.10(79.70, 281.95)µg/L vs. 79.60(42.83, 183.90)µg/L] in the high IS group were higher than those in the low IS group (P<0.05), and left ventricular ejection fraction [(45.15±10.65)% vs. (51.95±12.91)%] in the high IS group was lower than that in the low IS group (P<0.05). Multivariate Cox regression analyses showed that IS was independently associated with the risk of cardiac death in patients with acute STEMI two years after primary PCI(P=0.033, HR=1.075, 95%CI1.006-1.148). Every 1% increase in IS was associated with a 7.5% increase in cardiac death. Conclusions: Infarct size, measured by CMR within one week after primary PCI, is strongly associated with cardiac death in patients with acute STEMI two years after primary PCI. IS could be used as an index for the prognosis of patients with acute STEMI.

[The influence of XRCC1 gene polymorphism on the prognosis and safety of stage â ¢ patients with colorectal cancer who received oxaliplatin based adjuvant chemotherapy].

Objective: To investigate the influence of X-ray repair cross complementing 1 (XRCC1) gene polymorphism on the prognosis and safety of stage Ⅲ patients with colorectal cancer (CRC) who received oxaliplatin based adjuvant chemotherapy. Methods: A total of 218 stage Ⅲ patients with CRC after R0 resection and received oxaliplatin based adjuvant chemotherapy in the department of gastrointestinal surgery of the First Affiliated Hospital of Zhengzhou University from March 2012 to December 2019 were included and the baseline characteristics were collected. There were 125 male and 93 female patients, aged from 18 to 78 years. Peripheral blood and peripheral blood mononuclear cell (PBMC) specimens of the colorectal cancer patients were preserved for genotyping of XRCC1 gene genetic variation and mRNA expression of XRCC1, respectively. The association between genotype status and prognosis was analyzed by Kaplan-Meier survival analysis. And the correlation between genotype status and adverse reactions was performed with χ2 test. Results: The median follow-up time were 4.9 (0.3-7.3) years. The median disease-free survival (DFS) of the 218 patients with CRC was 4.4 years and the median overall survival (OS) was 5.5 years. The prevalence of rs1799782 in XRCC1 gene among the 218 patients was: GG genotype 62.4% (136/218), GA genotype 33.0% (72/218) and AA genotype 4.6% (10/218), minor allele frequency was 0.21. And the distribution frequencies of the three genotypes were in accordance with the hardy-weinberg equilibrium (P=0.905). GA and AA genotypes were merged in the subsequent analysis. The median DFS [M (95%CI)] of GG genotype and GA/AA genotype was 5.2 (4.5-5.9) years and 3.8 (3.2-4.4) years, which was statistically significant (χ²=6.943, P=0.008). Furthermore, the median OS [M (95%CI)] of the two genotypes were 6.0 (5.3-6.7) years and 4.5 (3.9-5.1) years, which was statistically significant (χ²=5.538, P=0.010). The mRNA expression of XRCC1 in PBMC of the patients with GA/AA genotypes was 3.8±0.6,which was significantly higher than that of the GG genotype patients(2.8±0.7) (t=6.140, P<0.001). Conclusion: The prognosis of patients with CRC who received oxaliplatin based adjuvant chemotherapy may be influenced by XRCC1 rs1799782 through mediating the mRNA expression of XRCC1.

Hollow Plasma Acceleration Driven by a Relativistic Reflected Hollow Laser.

In order to address the present difficulty in experimentally generating the relativistic Laguerre-Gaussian laser, primarily due to damage caused to optical modulators, a high-reflectivity phase mirror is applied in the femtosecond petawatt laser system to generate a relativistic hollow laser at the highest intensity of 6.3×10^{19} W/cm^{2} for the first time. A simple optical model is used to verify that the vortex laser may be generated in this new scheme; using such a relativistic vortex laser, the hollow plasma drill and acceleration are achieved experimentally and proven by particle-in-cell simulations. With the development of the petawatt laser, this scheme opens up possibilities for the convenient production of the relativistic hollow laser at high repetition and possible hollow plasma acceleration, which is important for a wide range of applications such as the generation of radiation sources with orbital angular momentum, fast ignition for inertial confinement fusion, and jet research in the astrophysical environment.

Long noncoding RNA NNT-AS1 functions as an oncogene in breast cancer via repressing ZFP36 expression.

Breast cancer is one of the most common cancers in women. This study focuses on the effects of Long non-coding RNAs (lncRNAs) NNT-AS1 on breast cancer cell growth and metastasis. Fifty-six pairs of breast cancer (BC) tissues and matched paracarcinoma tissues were obtained. The BC cell lines and normal human breast cell line were employed. NNT-AS1 in BC cells was knocked down by shRNA. Cell counting kit-8 assay (CCK-8), colony formation assay, cell cycle analysis, cell apoptosis analysis, cound healing assay, Transwell assay, cioinformatics analysis, Western blot analysis and Xenograft model were used. Quantitative real-time polymerase chain reaction (qRT-PCR) assay indicated that expression of NNT-AS1 was obviously upregulated in breast cancer tissues compared with adjacent tissues (n=56). Knockdown of NNT-AS1 could attenuate breast cancer cell viability, proliferation, invasion and migration, as well as promote cell apoptosis and induce cell cycle arrest at G0/G1 phase. ZFP36 was directly combined with NNT-AS1, and silencing of ZFP36 could rescue tumor suppression role by downregulating NNT-AS1 on cell proliferation and metastasis. Knockdown of NNT-AS1 could suppress cell growth and metastasis via interacting with ZFP36 in vivo. This study demonstrated that knockdown of NNT-AS1 had tumor-suppressive effect on breast cancer progression and metastasis via interacting with ZFP36 in vitro and in vivo, which provides a new insight into the treatment and prognosis evaluation of breast cancer.

[Features of gut microbiota in patients with idiopathic Parkinson's disease].

Objectives: To investigate whether the fecal microbiome of Parkinson's disease patients differs from that of healthy population and explore the pathogenesis and new treatment of idiopathic Parkinson's disease. Methods: A total of 30 patients diagnosed as idiopathic Parkinson disease (PD group) in Beijing Friendship Hospital between April 2017 and June 2018 were enrolled and 30 healthy controls (NC group) were recruited at the same time.Medical records and score of unified Parkinson's disease rating scale (UPDRS) were collected and fresh fecal samples were obtained and stored in refrigerator (-80℃). The microbial compositions of fecal samples were investigated by 16S rRNA gene sequencing targeting the V3-V4 region. The taxa abundance and microbial composition were tested. Results: There was no difference of age and sex in PD and NC groups. Chao1 and Shannon indexes tended to be higher in PD group, yet failed to reach statistic significance (P=0.115 and 0.052). Relative abundance of gut microbiota differed in each taxonomic category. The relative abundance of Firmicutes in PD group was 53.6%(41.7%-64.8%), while that of Bacteroidetes in NC group was 51.7%(31.7%-65.3%). The ratios of Firmicutes to Bacteroidetes were significantly different between the two groups (1.6(0.9-3.4) vs 0.7(0.5-1.4), P=0.001). In Clostridia, Bacilli and Erysipelotrichia of Firmicutes, the relative abundances of Clostridiales, Christensenellaceae, Peptoclostridium, Lactobacillus and Erysipelatoclostridium were higher in PD group (P=0.024, 0.046, 0.036, 0.022 and 0.037). The relative abundance of Prevotella of Bacteroidales, was lower in PD group, yet failed to reach statistic significance (P=0.121). The relative abundances of Alistipes of Rikenellaceae and Butyricimonas of Marinilabiliales in PD group were significantly higher than those in NC group (P=0.047 and 0.033). The relative abundance of Bifidobacterium of Actinobacteria was significanly higher in PD group when compared with NC group (P=0.009). Despite the relatively low abundance, Akkermansia of Verrucomicrobia was significantly higher in PD group than in NC group (P=0.025). Conclusion: The structures of the fecal microbiota differ significantly between PD patients and healthy controls.

Collisionless Shock Acceleration of High-Flux Quasimonoenergetic Proton Beams Driven by Circularly Polarized Laser Pulses.

We present experimental studies on ion acceleration using an 800-nm circularly polarized laser pulse with a peak intensity of 6.9×10^{19} W/cm^{2} interacting with an overdense plasma that is produced by a laser prepulse ionizing an initially ultrathin plastic foil. The proton spectra exhibit spectral peaks at energies up to 9 MeV with energy spreads of 30% and fluxes as high as 3×10^{12} protons/MeV/sr. Two-dimensional particle-in-cell simulations reveal that collisionless shocks are efficiently launched by circularly polarized lasers in exploded plasmas, resulting in the acceleration of quasimonoenergetic proton beams. Furthermore, this scheme predicts the generation of quasimonoenergetic proton beams with peak energies of approximately 150 MeV using current laser technology, representing a significant step toward applications such as proton therapy.

High-Brightness High-Energy Electron Beams from a Laser Wakefield Accelerator via Energy Chirp Control.

By designing a structured gas density profile between the dual-stage gas jets to manipulate electron seeding and energy chirp reversal for compressing the energy spread, we have experimentally produced high-brightness high-energy electron beams from a cascaded laser wakefield accelerator with peak energies in the range of 200-600 MeV, 0.4%-1.2% rms energy spread, 10-80 pC charge, and ∼0.2 mrad rms divergence. The maximum six-dimensional brightness B_{6D,n} is estimated as ∼6.5×10^{15} A/m^{2}/0.1%, which is very close to the typical brightness of e beams from state-of-the-art linac drivers. These high-brightness high-energy e beams may lead to the realization of compact monoenergetic gamma-ray and intense coherent x-ray radiation sources.

Transient-grating self-referenced spectral interferometry for infrared femtosecond pulse characterization.

We propose a technique for measuring infrared femtosecond pulses: transient-grating self-referenced spectral interferometry. Based on this technique, we built an extremely simple, alignment-free device and successfully characterized both 38 fs pulses at 800 nm and sub-two-cycle 10 fs pulses at 1.75 µm.

All-optical cascaded laser wakefield accelerator using ionization-induced injection.

We report on near-GeV electron beam generation from an all-optical cascaded laser wakefield accelerator (LWFA). Electron injection and acceleration are successfully separated and controlled in different LWFA stages by employing two gas cells filled with a He/O2 mixture and pure He gas, respectively. Electrons with a Maxwellian spectrum, generated from the first LWFA assisted by ionization-induced injection, were seeded into the second LWFA with a 3-mm-thick gas cell and accelerated to be a 0.8-GeV quasimonoenergetic electron beam, corresponding to an acceleration gradient of 187 GV/m. The demonstrated scheme paves the way towards the multi-GeV laser accelerators.

Heparin-functionalized collagen matrices with controlled release of basic fibroblast growth factor.

Tissue engineering scaffolds with controlled long-term release of growth factors are constructed in an attempt to mimic the intelligent ability of the extracellular matrix (ECM) to release endogenous growth factors. In this study, collagen sponges (Collagen group) were modified by N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) crosslinking (EDC/NHS group) and heparin immobilization (EDC/NHS-H group), and subsequently seeded with human umbilical vein endothelial cells (HUVECs). Native and modified sponges were pre-adsorbed with basic fibroblast growth factor (bFGF) to evaluate the sustained release and bioactive maintenance of bFGF from the sponges. We found that modified collagen matrices permitted HUVECs to proliferate and migrate well and to distribute uniformly. The EDC/NHS-H group exhibited an excellent sustained-release profile and bioactive maintenance of the pre-adsorbed bFGF as compared with the Collagen and EDC/NHS groups. These results suggest that heparin-functionalized collagen matrices can support a controlled release of bFGF and thus, have potential as a tissue engineering scaffold.

Clinical study and observation on the effect of hemoperfusion therapy treatment on central nervous system injury in patients with 2,4-dichlorophenoxyacetic acid poisoning.

OBJECTIVE: To evaluate the curative effect of hemoperfusion therapy on central nervous system injury in patients with 2,4-dichlorophenoxyacetic acid poisoning. PATIENTS AND METHODS: A total of 60 patients with 2,4-dichlorophenoxyacetic acid poisoning were enrolled in this study. They were admitted to the Emergency Department of Qinghai Provincial People's Hospital from 2015 to 2018 and were randomly divided into two groups by random number table method. One group was control group (routine treatment group), and the other group was the treatment group (hemoperfusion therapy was added on the basis of routine treatment). Glasgow coma score (GCS), APACHE II score, and MMSE score were used to evaluate the effects before treatment and 7 days after treatment. The results were statistically analyzed. RESULTS: After treatment, GCS in the treatment group was higher than that in the control group, while APACHE II score was lower than that in the control group, and MMSE score was significantly higher than that in the control group, with statistically significant difference (p<0.05). The effective rate in the control group was only 26.67%, and that in the treatment group was 86.67%, with statistically significant difference (c2=19.62, p<0.001). CONCLUSIONS: Hemoperfusion therapy can promote the recovery of central nervous system in patients with 2,4-dichlorophenoxyacetic acid poisoning, reduce the injury of other organs, and significantly reduce the mortality of patients.

[Advances in the application of three-dimensional photothermal functionalized scaffold for bone repairation after maxillofacial tumor surgery].

The traditional treatment of maxillofacial tumors includes surgery and chemoradiotherapy. Surgery carries a high risk,and large bone defects are difficult to repair themselves. Besides, chemoradiotherapy can bring serious side effects. Therefore, how to repair the bone defects after maxillofacial tumor surgery and prevent tumor recurrence has become a major clinical challenge. In recent years, photothermal therapy has attracted much attention because of its low invasion, high efficiency, and no side effects. More and more photothermal materials have emerged and been applied in photothermal therapy. In order to treat tumor-related bone defects, many studies use three-dimensional printing technology to prepare photothermal functionalized scaffold to repair bone defects and prevent the recurrence of tumors. Based on photothermal therapy, the review states the photothermal materials, bone repair materials and photothermal functionalized scaffolds in the treatment of maxillofacial tumors at home and abroad, and provides a new way to solve the clinical challenge of bone defect and tumor recurrence after maxillofacial tumor surgery.

[Trends of female reproductive health status in Bangladesh from 1999 to 2018].

Objective: To analyze the changes of relevant indicators in reproductive health status among Bangladeshi women from 1999 to 2018 and to assess whether the 2030 Sustainable Development Goals (SDGs) can be achieved. Methods: Data were obtained from both the Bangladesh Demographic and Health as well as from the Maternal Mortality and Health Care Surveys. The trends of SDGs indicators related to reproductive health from 1999 to 2018 were analyzed and compared, and the average annual rate of change was calculated. Development index was used to assess the difficulty of achieving the SDGs. Results: The maternal mortality rate increased first and then leveled off from 2001 to 2016. From 1999 to 2018, the coverage of reproductive health care services and the proportion of women who had the right to make the decision on their own health care service, were generally increasing. Proportion of the following areas as: "contraceptive needs, women who consider that partner violence is justified, the rate of early marriage, and the rate of early childbearing etc.", were declining at various degrees. Development index of the antenatal care coverage, rate of delivery in medical facilities, percentage of live births attended by medically trained providers and the rate of postnatal care etc., were less than 1. The development indices of the maternal mortality rates were close to 1, while all the other indicators were greater than 1. Conclusions: The reproductive health-related SDGs indicators in Bangladesh appeared somehow degrees of progress from 1999 to 2018. However, for most indicators, the average annual rate of change was still lower than the expected to achieve the 2030 target which called for acceleration in the next few years.

MTIM affects retinal ganglion cells through PI3K/AKT signaling pathway.

OBJECTIVE: Retinal ganglion cells (RGCs) are the main cells that form vision in the retina. MT1M is involved in the occurrence and progression of various diseases. However, the role of MTIM in RGCs cells remains unclear. MATERIALS AND METHODS: RGCs were cultured in vitro and randomly divided into control group, MT1M group (transfected with MT1M-pcDNA3.1 plasmid), and MT1M siRNA group (transfected with MT1M siRNA) followed by measuring MT1M and NT-3 expression by real time PCR and Western blot, cell proliferation by MTT assay, secretion of IL-2 and IL-6 by enzyme-linked immunosorbent assay (ELISA), SOD activity and ROS content. In addition, expression of PI3K/AKT signaling pathway protein was detected by Western blot. RESULTS: MT1M expression in MT1M group was significantly increased, which promoted cell proliferation, increased NT-3 expression, and decreased Caspase 3 activity and IL-2 and IL-6 secretion. Meanwhile, SOD activity was increased, ROS content was decreased and PI3K/AKT protein phosphorylation was elevated. The differences were statistically significant compared with control group (p < 0.05). MT1M siRNA decreased MT1M expression, inhibited cell proliferation, decreased NT-3, and increased Caspase 3 activity and IL-2 and IL-6 secretion. In addition, MT1M siRNA decreased SOD activity, increased ROS content and reduced PI3K/AKT protein phosphorylation. Compared with control group, the differences were statistically significant (p < 0.05). CONCLUSIONS: Up-regulation of MT1M can inhibit RGC cell apoptosis and inflammation, and promote RGC cell proliferation through the PI3K/AKT signaling pathway.

MiR-204 inhibits inflammation and cell apoptosis in retinopathy rats with diabetic retinopathy by regulating Bcl-2 and SIRT1 expressions.

OBJECTIVE: To explore the influences of micro ribonucleic acid (miR)-204 on the rats with diabetic retinopathy by regulating the expressions of B-cell lymphoma 2 (Bcl-2) and sirtuin 1 (SIRT1). MATERIALS AND METHODS: A total of 36 Sprague-Dawley rats were randomly assigned into normal group (n=12), model group (n=12), and miR-204 mimics group (n=12). No treatment was performed in the normal group, the diabetic retinopathy model was established in model group, and miR-204 mimics were administered for intervention after modeling in the inhibitor group. After 7 d, materials were sampled for detection. The expressions of Bcl-2 and SIRT1 were detected via immunohistochemistry, and their relative protein expression levels were determined via Western blotting (WB). Quantitative Polymerase Chain Reaction (qPCR) was performed to detect the expression of miR-204, and the content of inflammatory factors interleukin (IL)-6, IL-18, and tumor necrosis factor-α (TNF-α) was measured using enzyme-linked immunosorbent assay (ELISA). Finally, cell apoptosis was evaluated via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). RESULTS: Immunohistochemistry results showed that the positive expression levels of Bcl-2 and SIRT1 were substantially lower in the model and miR-204 mimics groups than those in the normal group (p<0.05), and their positive expression levels in miR-204 mimics group were notably higher than those in model group (p<0.05). According to Western blot (WB) results, the relative protein expression levels of Bcl-2 and SIRT1 markedly declined in the other two groups compared with those in the normal group (p<0.05), while miR-204 mimics group exhibited remarkably higher relative protein expression levels of Bcl-2 and SIRT1 than the model group (p<0.05). The results of qPCR revealed that the relative expression level of miR-204 was markedly lowered in model and miR-204 mimics groups compared with that in the normal group (p<0.05), and its relative expression level in miR-204 mimics group was remarkably higher than that in the model group. It was found through enzyme-linked immunosorbent assay (ELISA) that compared with normal group, the other two groups had substantially increased content of IL-6, IL-18, and TNF-α (p<0.05), and the content of IL-6, IL-18, and TNF-α in miR-204 mimics group was markedly lower than that in the model group (p<0.05). According to TUNEL results, the apoptosis rate of cells rose substantially in the other two groups compared with that in the normal group (p<0.05), while was notably lower in the miR-204 mimics group than that in the model group (p<0.05). CONCLUSIONS: MiR-204 up-regulates Bcl-2 and SIRT1 expressions to inhibit the inflammation and cell apoptosis in rats with diabetic retinopathy.

Attenuating effect of melatonin on lipopolysaccharide-induced chicken small intestine inflammation.

Enriched melatonin (MEL) has been found in the mammalian intestine and has been recently demonstrated to alleviate rodent colitis. In this study, the effect of MEL on lipopolysaccharide (LPS)-induced intestinal inflammations was investigated in new chicken hatchlings. The chicks were fed with a diet supplemented with MEL (12.5 mg/day) from D1 to D10. Meanwhile, the chicks in the LPS or MEL + LPS groups were injected with LPS (10 mg/kg BW, i.p.) at D10. LPS treatment for 6 h increased the expression of IL-6, IL-4, caspase-3 mRNAs and TUNEL-positive cell populations, but decreased populations of the goblet and PCNA+ cells, IgA production and the expression of MUC2 mRNA in the duodenum. Compared with the LPS group, MEL pre-feeding alleviated duodenal inflammation and decreased the expression of TNF-α mRNAs by 23.6% (P = 0.004), IL-6 mRNAs by 69.4% (P = 0.001), IL-4 mRNAs by 4.1% (P = 0.824) and caspase-3 mRNAs by 45.8% (P < 0.001). Conversely, MEL pre-feeding attenuated the LPS-induced changes of IgA production by 161.6% (P = 0.013) and PCNA+ cell populations by 172.1% (P < 0.001) in the duodenum. TLR4 mRNA was also up-regulated by LPS treatment but down-regulated by MEL pre-feeding. In conclusion, dietary MEL could attenuate LPS-induced chick duodenal inflammation by down-regulating the expression of inflammatory cytokines, promoting epithelial cell proliferation, improving the immunological barrier and inhibiting epithelial apoptosis via the mediation of TLR4.

Culture and characterization of chicken small intestinal crypts.

The integrity and normal function of the small intestinal epithelium depends critically on the rapid renewal of epithelial cells from basal stem cells. The intensive proliferation that fuels this self-renewal process is confined to the intestinal crypts. Establishment of suitable protocols for crypt isolation and culture is pivotal for the studies of intestinal self-renewal mechanisms. In this study, chicken small intestinal crypts were isolated, purified, and further cultured in a Matrigel 3-D culture system. The growth factor concentration assay on the fourth d of culture showed that Group C (50 ng/mL epidermal growth factor (EGF), 100 ng/mL Noggin, and 500 ng/mL R-spondin 1) supplement in culture medium could significantly enlarge the diameter of organoids when compared with Group A (5 ng/mL EGF, 10 ng/mL Noggin, 50 ng/mL, and R-spondin 1) and Group B (10 ng/mL EGF, 20 ng/mL Noggin, and 100 ng/mL R-spondin 1) by 188.4% (P = 0.026) and 176.9% (P = 0.034), respectively. Transmission electron microscopy, neutral red staining, and 5-ethynyl-2΄-deoxyuridine incorporation demonstrated the integrated structure, high viability, and proliferative activity in cultured chicken intestinal organoids. In addition, intestinal stem cell marker genes (Olfm4, Znrf3, Hopx, and Lgr5) also could be detected in cultured intestinal organoids. Furthermore, CHIR99021 (a glycogen synthase kinase 3ß inhibitor) could enhance the expression of Olfm4, Znrf3, Hopx, and Lgr5 by 750% (P = 0.001), 467% (P < 0.001), 450% (P < 0.001), and 333% (P = 0.008), respectively, indicating the responsiveness of the cultured chicken intestinal organoids to exogenous stimulus. This study modified a murine culture model and optimized it to provide a chicken intestinal organoid model for use as a physiological or pathological research platform in vitro.