ABSTRACT
BACKGROUND@#Current replacement procedures for stenosis or occluded arteries using prosthetic grafts have serious limitations in clinical applications, particularly, endothelialization of the luminal surface is a long-standing unresolved problem.METHOD: We produced a cell-based hybrid vascular graft using a bioink engulfing adipose-derived mesenchymal stromal cells (ADSCs) and a 3D bioprinting process lining the ADSCs on the luminal surface of GORE-Tex grafts. The hybrid graft was implanted as an interposition conduit to replace a 3-cm-long segment of the infrarenal abdominal aorta in Rhesus monkeys. @*RESULTS@#Complete endothelium layer and smooth muscle layer were fully developed within 21 days post-implantation, along with normalized collagen deposition and crosslinking in the regenerated vasculature in all monkeys. The regenerated blood vessels showed normal functionality for the longest observation of more than 1650 days. The same procedure was also conducted in miniature pigs for the interposition replacement of a 10-cm-long right iliac artery and showed the same long-term effective and safe outcome. @*CONCLUSION@#This cell-based vascular graft is ready to undergo clinical trials for human patients.
ABSTRACT
Acute pancreatitis （AP） is a common clinical acute abdominal disease， which is characterized by acute onset， rapid development， severe disease， many complications， and high mortality rate. It can progress to severe AP （SAP） if not treated promptly in the early stage. The pathogenesis of AP is complex and involves multiple cellular and molecular levels. It is now clear that oxidative stress and reactive oxygen species （ROS） production are involved in the physiopathological process of AP， which is associated with a low quantity and activity of antioxidant enzymes in pancreatic cells. Nuclear factor E2-related factor 2 （Nrf2） serves as the ''golden key'' to maintain redox homeostasis in tissue cells and constitutes an important signaling pathway for antioxidant response and inflammation in vivo by collaborating with downstream antioxidant enzymes such as heme oxygenase-1 （HO-1）. Traditional Chinese medicine has unique efficacy in treating diseases due to its multi-component， multi-target， multi-drug delivery， and multi-formulation characteristics. Based on the concept of synergy between traditional Chinese and Western medicine， traditional Chinese medicine is becoming a new craze in the treatment of AP. The level of oxidative stress and Nrf2/HO-1 signaling pathway in AP pancreatic tissue are in a dynamic change process， and the intervention of traditional Chinese medicine can clean ROS production， affect the inflammatory pathway， and reduce oxidative stress damage， so as to protect against pancreatic injury. This suggests that this pathway plays an important role in AP. This article reviews the recent literature on the regulation of the Nrf2/HO-1 signaling pathway by traditional Chinese medicine for AP and summarizes that the monomers of traditional Chinese medicine targeting this pathway are mainly heat-clearing and detoxifying， blood-activating and blood-stasis-removing， and Qi benefiting and middle warming， and the compounds of traditional Chinese medicine include Yinchenhao Decoction and QingYi Ⅱ， so as to provide a new direction for the prevention and treatment of AP and further drug development.
ABSTRACT
Objective:We applied a hypoxia-induced model of human fetal retinal microvascular endothelial cell (RMEC) to study the effect of carbonic anhydrase 9 (CA9) on cell proliferation.Methods:The eyeballs of spontaneously aborted fetuses in Guangdong Women and Children's Hospital were obtained, and the retinas were isolated. RMEC was obtained by trypsin and collagenase two-step enzyme digestion, and endothelial cells were identified by CD34. The fetal RMEC and the purchased adult RMEC were cultured in normoxic and hypoxic incubators (1%O 2+5%CO 2+94%N 2), and the expression of CA9 was detected by qPCR and Western blot. After knocking down the CA9 by small interference RNA technique, the cell proliferation was detected by CCK-8 method, and the cell viability was detected by CCK-8 after adding CA9 inhibitor U-104. Results:The primary RMEC was extracted successfully. Immunofluorescence staining showed the percentage of CD34 positive cells in the third-generation cells was nearly 100%. The expression of CA9 mRNA in immature fetus and adult RMEC under hypoxia culture was higher than that under normoxic culture (fetal 1% O 2 group vs. fetal 21% O 2 group: 67.80±10.31 vs. 1.00±0.04, P<0.001; adult 1% O 2 group vs. adult 21% O 2 group: 1.72±0.22 vs. 1.00±0.02, P=0.014). Western blot analysis showed significantly increased expression of CA9 in the fetal RMEC exposed to hypoxia, which aligned with the expression of CA9 mRNA. When fetal RMEC was transfected with siCA9 20 nM, the knockdown rate of CA9 was 95% ( P<0.001). CCK-8 assay showed significantly lower proliferation of fetal RMEC cells in siCA9 group compared to siNC group (0.57±0.05 vs. 0.90±0.03, P<0.001), which was reflected by the OD value. With the addition of 100 μM CA9 inhibitor U-104, the viability of fetal RMEC in the treated groupwas significantly lower than that in the untreated group (99.16%±3.82% vs. 119.10% ±1.72%, P=0.002). Conclusions:The expression of CA9 differed between adult and preterm fetus in our hypoxia-induced RMEC model. Inhibiting CA9 can inhibit the proliferation of retinal microvascular endothelial cells of preterm fetus.
ABSTRACT
The Asian expert working group completed the 2022 consensus on vernal keratitis (VKC), which provides detailed information on the definition, classification, clinical manifestation, diagnosis and treatment of VKC in Asia and identifies unmet treatment needs, including: (1) lack of clear criteria for the diagnosis and treatment of patients with VKC.The pathogenesis of the disease is unclear, and anti-allergy therapy is often ineffective in patients with moderate or severe VKC; (2) the need for standardized treatment and management strategies.Treatment and management of patients with the same severity varies widely among Asian countries, with a lack of clear, standardized guidelines and grading systems; (3) safety and iatrogenic complications; (4) optimal dosing regimen for pharmacologic treatment.In this article, the aspects mentioned above were interpreted and analyzed in the context of the Expert Consensus on the Diagnosis and Treatment of Allergic Conjunctivitis in China ( 2018), with a view to helping the prevention and treatment of VKC in China.
ABSTRACT
ObjectiveTo investigate the therapeutic effect of Qingjie Huagong decoction （QJHGD） on a mouse model of severe acute pancreatitis （SAP） and the mechanism of action of QJHGD against inflammatory response. MethodsA total of 36 male C57BL/6J mice were randomly divided into blank group， model group， Western medicine group （ulinastatin）， and low-， middle-， and high-dose QJHGD groups， with 6 mice in each group. All mice except those in the blank group were given 5% sodium taurocholate by retrograde pancreaticobiliary injection to establish a model of SAP. After modeling， the mice in the low-， middle-， and high-dose groups were given QJHGD （1， 2， and 4 g/kg， respectively） by gavage， and those in the Western medicine group were given intraperitoneal injection of ulinastatin （5×104 U/kg）， for 7 days in total. HE staining was used to observe the histopathological changes of the pancreas； ELISA was used to measure the levels of α-amylase， lipase， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-18 （IL-18）， and tumor necrosis factor-α （TNF-α） in mice； RT-qPCR was used to measure the mRNA expression levels of NOD-like receptor protein3 （NLRP3）， Toll-like receptor 4 （TLR4）， and nuclear factor-kappa B （NF-κB） in pancreatic tissue； immunohistochemistry was used to measure the positive expression rates of NLRP3， TLR4， and NF-κB in pancreatic tissue； Western blot was used to measure the protein expression levels of NLRP3， TLR4， NF-κB， IL-1β， and IL-6. An analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the blank group， the model group had diffuse destruction of pancreatic tissue structure， focal dilatation of pancreatic lobular septum， pancreatic acinar atrophy， and massive inflammatory cell infiltration， as well as significant increases in the content of α-amylase， lipase， IL-1β， IL-6， IL-8， IL-18， and TNF-α （all P<0.05）， the mRNA expression levels and positive expression rates of NLRP3， TLR4， and NF-κB （all P<0.05）， and the protein expression levels of NLRP3， TLR4， NF-κB， IL-1β， and IL-6 （all P<0.05）. Compared with the model group， the low-， middle-， and high-dose QJHGD groups and the Western medicine group had slightly tighter and more intact structure of pancreatic tissue， ordered arrangement of pancreatic acinar cells， a small amount of inflammatory cell infiltration， and hemorrhagic foci of pancreatic lobules， as well as significant reductions in the content of α-amylase， lipase， IL-1β， IL-6， IL-8， IL-18， and TNF-α （all P<0.05）， the mRNA expression levels and positive expression rates of NLRP3， TLR4， and NF-κB （all P<0.05）， and the protein expression levels of NLRP3， TLR4， NF-κB， IL-1β， and IL-6 （all P<0.05）. ConclusionQJHGD may exert a protective effect on the pancreatic tissue of SAP mice by inhibiting the activation of NLRP3/TLR4/NF-κB signaling pathway-related proteins， reducing the release of inflammatory mediators， and preventing the enhancement of inflammatory cascade response.
ABSTRACT
Objective:To study the feasibility and value of applying organ dose modulation technique in computed tomography(CT)scanning on eyes.Methods:A total of 330 patients who admitted to Beijing Friendship Hospital affiliated with Capital Medical University from August 2021 to August 2023 and underwent CT examination on eyes were selected.They were divided into modulation group,conventional group and low-dose group based on the control methods of tube current in scanning.The modulation group used organ dose modulation technique with intelligent automatic tube current of 70-150 mA,and the conventional dose group used a fixed tube current of 100 mA,and the low-dose group used a fixed tube current of 70 mA.The differences in radiation dose and CT imaging quality among 3 groups were compared.Results:The dose length product(DLP),volume CT dose index(CTDIvol)and effective dose(ED)of cross-sectional scan of the modulation group were respectively 62.37,12.83 and 0.143 mSv,which were significantly lower than 115.23,18.93 and 0.265mSv of conventional dose group,respectively,with statistically significant(F=2544.944,6009.596,2544.944,P<0.05).The DLP,CTDIvol and ED of coronary scan of the modulation group were 68.19,13.15 and 0.156,respectively,which were significantly lower than 122.41,19.20 and 0.282 mSv of the conventional dose group,respectively,with statistically significant(F=1232.413,3813.940,1232.413,P<0.05).The excellent rates of cross-sectional scans among the modulation group,conventional group and lower-dose group were respectively 98.83%,86.15%and 47.69%,and the difference among three groups was statistically significant(x2=53.908,P<0.05).The excellent rates of image qualities of the modulation group,conventional group and low-dose group were respectively 96.61%,80.70%and 48.28%,and the difference among three groups was statistically significant(x2=20.992,P<0.05).Conclusion:The application of organ dose modulation technique in CT scanning on eyes can significantly reduce radiation dose on patient under ensures imaging quality can meet diagnostic needs when undergoes cross-sectional and coronal scans,which has higher clinical feasibility and application value.
ABSTRACT
Background The benchmark dose (BMD) method calculates the dose associated with a specific change in response based on a specific dose-response relationship. Compared with the traditional no observed adverse effect level (NOAEL) method, the BMD method has many advantages, and the 95% lower confidence limit of benchmark dose lower limit (BMDL) is recommended to replace NOAEL in deriving biological exposure limits. No authority has yet published any health-based guideline for rare earth elements. Objective To evaluate genotoxicity threshold induced by acute exposure to neodymium nitrate in mice using BMD modeling through micronucleus test and comet assay. Methods SPF grade mice (n=90) were randomly divided into nine groups, including seven neodymium nitrate exposure groups, one control group (distilled water), and one positive control group (200 mg·kg−1 ethyl methanesulfonate), 10 mice in each group, half male and half female. The seven dose groups were fed by gavage with different concentrations of neodymium nitrate solution (male: 14, 27, 39, 55, 77, 109, and 219 mg·kg−1; female: 24, 49, 69, 97, 138, 195, and 389 mg·kg−1) twice at an interval of 21 h. Three hours after the last exposure, the animals were neutralized by cervical dislocation. The bone marrow of mice femur was taken to calculate the micronucleus rate of bone marrow cells, and the liver and stomach were taken for comet test. Results The best fitting models for the increase of polychromatophil micronucleus rate in bone marrow of female and male mice induced by neodymium nitrate were the exponential 4 model and the hill model, respectively. The BMD and the BMDL of female mice were calculated to be 31.37 mg·kg−1 and 21.90 mg·kg−1, and those of male mice were calculated to be 58.62 mg·kg−1 and 54.31 mg·kg−1, respectively. The best fitting models for DNA damage induced by neodymium nitrate in female and male mouse hepatocytes were the exponential 5 model and the exponential 4 model, respectively, and the calculated BMD and BMDL were 27.15 mg·kg−1 and 11.99 mg·kg−1 for female mice, and 16.28 mg·kg−1 and 10.47 mg·kg−1 for male mice, respectively. The hill model was the best fitting model for DNA damage of gastric adenocytes in both female and male mice, and the calculated BMD and BMDL were 36.73 mg·kg−1 and 19.92 mg·kg−1 for female mice, and 24.74 mg·kg−1 and 14.08 mg·kg−1 for male mice, respectively. Conclusion Taken the micronucleus rate of bone marrow cells, DNA damage of liver cells and gastric gland cells as the end points of genotoxicity, the BMDL of neodymium nitrate is 10.47 mg·kg−1, which can be used as the threshold of genotoxic effects induced by acute exposure to neodymium nitrate in mice.
ABSTRACT
Objective To investigate the clinical efficacy and safety of long-term intermittent oral ad-ministration of fosfomycin trometamol(FMT)in the control of urinary tract infection and the reduction of stone recurrence rate after removal of upper urinary infection stones.Methods A total of 171 patients who met the inclusion criteria were enrolled and divided into the FMT group(using FMT),cephalosporin group(using cefixime),and blank group(not using antibiotics)according to the random number method,with 57 cases in each group.Finally,55 cases in the FMT group,47 cases in the cephalosporin group and 48 cases in the blank group were included in the statistical analysis,and the urinary tract infection and stone recurrence of the three groups were followed up regularly after the stone removal operation.Results There was no statisti-cal significance in the baseline data of the three groups(P>0.05).There were significant differences in the recurrence rate of urinary tract infection at the 3rd and 6th month among the 3 groups(P=0.010,P<0.001).Further pair-wise comparison showed that the recurrence rate of urinary tract infection at the 3rd month in the FMT group was lower than that in the blank group(P<0.05),but there was no statistical difference compared with the cephalosporin group(P>0.05).The recurrence rate of urinary tract infection at the 6th month in the FMT group was lower than that in the cephalosporin group and blank group(P<0.05).The recurrence rate of stones in the 1st and 3rd year of the three groups were statistically different(P= 0.028,0.015).Further pair-wise comparison showed that the 1st year stone recurrence rate of the FMT group was lower than that of the cephalosporin group and blank group(P<0.05).The 3rd year stone recurrence rate of the FMT group was lower than that of the blank group(P<0.05),but there was no statistical difference compared with the cephalosporin group(P>0.05).There was no significant difference in the total incidence of adverse drug re-action between the FMT group and cephalosporin group(P=0.131).Conclusion FMT is superior to cephalospo-rin in the control of urinary tract infection after lithotripsy for upper urinary tract infection.
ABSTRACT
When refractory diarrhea comes on,it greatly affects the life and daily work of patients,and there is no unified treatment.Patients with refractory diarrhea have varying degrees of intestinal flora disorder,so rebuilding the intestinal micro ecosystem may be an effective way to treat refractory diarrhea.Fecal microbiota transplantation(FMT)has the potential to be an effective treatment for refractory diarrhea as a therapy that reconstructs normal intestinal flora.In recent years,FMT has been applied to the treatment of some refractory diarrhea related to intestinal flora imbalance,such as recurrent clostridium difficile infection,inflammatory bowel disease,irritable bowel syndrome,and has achieved good results,but some problems have not been properly solved so far.This article reviews the mechanism of action of FMT in the treatment of refractory diarrhea,its clinical application,research progress and current problems.
ABSTRACT
Objective To observe the protective effects of codonopsis pilosulae polysaccharide on lung tissues in mice with acute lung injury(ALI)induced by lipopolysaccharide(LPS)and its mechanism.Methods Fifty male Kunming mice were randomly divided into control group,model group,dexamethasone group,codonopsis polysaccharide high-dose group(300 mg/kg)and codonopsis polysaccharide low-dose group(150 mg/kg).The ALI model was established by intraperitoneal injection of LPS.All mice were given gavage administration according to the grouping except for the control group.0.3 s force expiratory volume(FEV 0.3)and force spirometry(FVC)and their ratios were measured using multiparametric lung function test system.The histopathology change of mouse lung was detected by hematoxylin-eosin(HE)staining,and the classification and count of inflammatory cells in alveolar lavage fluid(BALF)was detected by Richter-Giemsa staining.Levels of IL-1β,IL-6,MPO and TNF-α were measured by ELISA in BALF,and Western blot was used to detect the protein expression level of p-p38,p-IκB-α and p-p65.Results Compared with those in the control group,lung histopathological damage was more pronounced in the model mice,with significantly lower FEV 0.3,FVC,FEV0.3/FVC assay value,but signifi-cantly higher lung tissue wet mass/dry mass(W/D),neutrophils,lymphocytes,IL-1β,IL-6,MPO,TNF-α,p-p38 MAPK,p-IκB-α,and p-p65(P<0.05);while codonopsis pilosulae polysaccharide could significantly reverse these effects.Conclusion Codonopsis pilosulae polysaccharide plays a protective role against LPS-induced ALI via inhibiting MAPK/NF-κB pathway to reduce the pathological damage of lung tissue,and the level of inflammatory factors,thus to improve lung function in ALI model mice.
ABSTRACT
Objective To investigate the application value of single nucleotide polymorphism(SNP)linkage analysis based on next-generation sequencing(NGS)technology in preimplantation genetic testing(PGT)of families with autosomal recessive polycystic kidney disease(ARPKD).Methods A family with ARPKD was selected,where the female member had a pregnancy ultrasound revealing polycystic kidney in the fetus.Genetic testing showed compound heterozygous mutations of the polycystic kidney/polycystic liver disease 1 gene(PKHD1),c.10444C>T(paternal)and c.4303del(maternal),with the c.4303del mutation being reported for the first time.Targeting the coding region of the PKHD1 gene,335 high-density tightly linked SNP sites were selected in the upstream and downstream 2M regions using multiplex polymerase chain reaction(PCR)and NGS.The couple′s SNP risk haplotypes carrying gene mutations were constructed.After in vitro fertilization,blastocyst culture was performed.Trophoblastic cells obtained from the biopsy were subjected to whole-genome amplification,and NGS was used for linkage analysis and low-depth chromosomal aneuploidy screening of the embryos.Sanger sequencing was used to verify the results of embryo linkage analysis.Results Among the 6 biopsied embryos,4 were mutation-free and euploid,1 exhibited heterozygous for the mutation and mosaic while another unstable sequencing data,making it impossible to judge.One of the mutation-free and developmentally healthy euploid embryos was implanted into the maternal uterus,resulting in the full-term delivery of a healthy baby.Conclusion Application of NGS-based SNP linkage analysis in PGT can effectively blocking the vertical transmission of ARPKD within families,while avoiding abortion issues caused by aneuploid embryos.This study is also the first PGT report target-ing the PKHD1 gene c.4303del mutation.
ABSTRACT
BACKGROUND:Unilateral biportal endoscopic technique has been widely used in lumbar interbody fusion in recent years,but there is little comparison between its clinical efficacy and that of minimally invasive transforaminal lumbar interbody fusion(MIS-TLIF)in the treatment of lumbar degenerative disease,whether the unilateral biportal endoscopic technique is a safe and effective lumbar fusion remains to be further demonstrated. OBJECTIVE:To compare the clinical efficacy of unilateral biportal endoscopic lumbar interbody fusion(UBE-LIF)and MIS-TLIF in the treatment of lumbar degenerative diseases and explore a more efficient lumbar fusion procedure. METHODS:Patients with single-level lumbar degenerative disease were enrolled in Affiliated Hospital of Guilin Medical College from October 2020 to February 2022,including 35 patients who underwent UBE-LIF and 286 patients who underwent MIS-TLIF.Propensity score matching was used to eliminate confounders.Four covariates including sex,age,disease type and surgical segment were matched 1:1(caliper value 0.01).After matching,29 patients from each group were included in the study.The perioperative operative time,hemoglobin loss and hospital stay were compared between the two groups.Visual analog scale score and Oswestry disability index were used to evaluate the functional recovery of the two groups before,1,6 months and 1 year after operation.The excellent and good rate of the two groups was evaluated by the modified MacNab standard at the last follow-up.The fusion of the two groups was evaluated by Lenke Dynamic X-ray film. RESULTS AND CONCLUSION:(1)The operative time in the MIS-TLIF group was shorter than that in the UBE-LIF group(P＜0.05).The amount of intraoperative hemoglobin loss in the MIS-TLIF group was higher than that in the UBE-LIF group.The hospital stay in the MIS-TLIF group was longer than that in the UBE-LIF group,and the differences were statistically significant(P＜0.05).(2)The visual analog scale scores for lumbago and leg pain,and Oswestry disability index were significantly reduced in both groups 1,6 months,and 1 year after surgery compared to before surgery(P＜0.05).Except for the visual analog scale score for lumbago at 1 month after surgery,there was no significant difference in the visual analog scale score for lumbago and leg pain,and Oswestry disability index between the two groups at the above time points(P＞0.05).(3)At the last follow-up,the modified MacNab standard efficacy evaluation showed that the excellent and good rates were 93%(27/29)in the UBE-LIF group and 90%(26/29)in the MIS-TLIF group;there was no significant difference between the two groups(P＞0.05).(4)Lenke dynamic radiographic evaluation system evaluation for lumbar fusion exhibited that the fusion rate was 90%(grade A,21 cases;grade B,5 cases;grade C,3 cases)in the UBE-LIF group;the fusion rate was 86%(grade A,20 cases;grade B,5 cases;grade C,4 cases)in the MIS-TLIF group;there was no significant difference between the two groups(P＞0.05).(5)It is indicated that UBE-LIF and MIS-TLIF have similar clinical effects in the treatment of single-level lumbar degenerative disease with the advantages of less trauma,less bleeding and shorter hospital stay.In addition,the early postoperative lumbago was relatively mild and the learning curve was relatively smooth.Although the operative time in the UBE-LIF group was longer than that in the MIS-TLIF group,it was still a safe and effective operation.
ABSTRACT
Objective: To investigate the effect of hsa_circ_0000392 (circ_0000392) on the radiosensitivity of cervical cancer cells and explore its potential mechanism. Methods: Cervical cancer tissues and adjacent normal tissues of 42 patients with cervical cancer who were confirmed pathologically for the first time in Huaihe Hospital of Henan University from 2016 to 2019 were collected. According to the patients' response to radiotherapy, the cancer tissues were divided into radio-sensitive tissues and radio-resistant tissues. The expressions of circ_0000392, miR-145-5p, and CRKL in radiation-sensitive, radiation-resistant cervical cancer tissues and Hela, SiHa cells were detected by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and western blot. SiRNA circ_0000392, miR-145-5p mimic, miR-145-5p inhibitor, pcDNA 3.1-CRKL and its negative control were transfected into HeLa and Siha cells, respectively. After radiation induction, the survival fraction of cells was detected by clone formation assay, apoptosis was detected by flow cytometry, and the expressions of apoptosis-related proteins Bax and Bcl-2 and ERK pathway protein p-ERK1/2 and ERK1/2 were detected by western blot. The targeting relationship between circ_0000392, miR-145-5p and CRKL was verified by dual luciferase reporter gene assay. The effect of circ_0000392 on radiotherapy sensitivity of cervical cancer in vivo was observed in the tumor formation experiment in nude mice. Results: circ_0000392 and CRKL were upregulated in radiation-resistant tissues and cancer cells of cervical cancer, while miR-145-5p was downregulated. The clone formation numbers of Hela and SiHa cells in si-circ_0000392#1+ 6 Gy group were (78.67±10.97) and (71.00±9.54), respectively, which were lower than those in si-Ctrl+ 6 Gy group [(176.00±22.27) and (158.33±17.56), respectively]. The apoptosis rates were (41.55±3.40)% and (31.41±3.29)%, respectively, which were higher than those in si-Ctrl+ 6 Gy group [(15.91±1.37)% and (13.70±1.89)%, P<0.05]. The protein expression of Bax was higher than that of si-Ctrl+ 6 Gy group, and the protein expressions of Bcl2 was lower than those of si-Ctrl+ 6 Gy group. The clone formation numbers of Hela and SiHa cells in si-circ_0000392#1+ miR-145-5p inhibitor+ 6 Gy group were (171.33±25.01) and (137.00±21.66), higher than those in si-circ_0000392#1+ inhibitor NC+ 6 Gy group [(84.67±17.79) vs (71.00±11.00), P<0.05]. The apoptosis rates were (17.41±2.58) % and (15.96±1.25) %, lower than those of si-circ_0000392 #1+ inhibitor NC+ 6 Gy [(40.29±2.92)% and (30.82±2.34)%, respectively, P<0.05]. The expression of Bax protein was lower than that of si-circ_0000392#1+ inhibitor NC+ 6 Gy group, and the expressions of Bcl2 protein were higher than those of si-circ_0000392#1+ inhibitor NC+ 6 Gy group. Circ_0000392 can target miR-145-5p, and CRKL is the downstream target gene of miR-145-5p. The clone formation numbers of Hela and SiHa cells in miR-145-5p mimic+ 6 Gy group were (74.33±10.02) and (66.00±12.17), respectively, which were lower than those of mimic NC+ 6 Gy group [(197.67±17.21) vs (157.67±11.59), respectively, P<0.05]. The apoptosis rates were (45.58±2.16)% and (32.10±3.55)%, higher than those of mimic NC+ 6 Gy group [(15.85±2.45)% and (13.99±1.69)%, respectively, P<0.05]. The expression of Bax protein was higher than that of the mimic NC+ 6 Gy mimic group, and the expression of Bcl2 protein was lower than that of the mimic NC+ 6 Gy group. The clone formation numbers of Hela and SiHa cells in miR-145-5p mimic+ pcDNA-CRKL+ 6 Gy group were (158.00±15.88) and (122.33±13.65), respectively, which were higher than those of miR-145-5p mimic+ pcDNA+ 6 Gy group [(71.33±8.02) vs (65.67±12.22), P<0.05]. The apoptosis rates were (19.50±3.45)% and (17.04±0.94)%, respectively, which were lower than those of miR-145-5p mimic+ pcDNA+ 6 Gy group [(44.33±2.36)% and (32.05±2.76)%, respectively, P<0.05]. The expression of Bax protein was lower than that of miR-145-5p mimic+ pcDNA group+ 6 Gy group, and the expression of Bcl2 protein was higher than that of miR-145-5p mimic+ pcDNA+ 6 Gy group. Sh-circ_0000392 group had smaller tumor volume and decreased tumor weight (P<0.05). The relative mRNA expression levels of circ_0000392, miR-145-5p and CRKL and the relative protein expression levels of CRKL, Bcl-2 and p-ERK1/2 were decreased, while the relative expression level of Bax protein was increased (P<0.05). Conclusion: Circ_0000392 could enhance the radiosensitivity of cervical cancer cells, and its mechanism may be related to the regulation of CRKL/ERK signaling pathway by targeting miR-145-5p, which provides a new reference for enhancing the radiosensitivity of cervical cancer cells.
Subject(s)
Animals , Mice , Female , Humans , Uterine Cervical Neoplasms/radiotherapy , bcl-2-Associated X Protein/genetics , Mice, Nude , Proto-Oncogene Proteins c-bcl-2/genetics , Apoptosis , MicroRNAs/genetics , Cell Proliferation , Cell Line, TumorABSTRACT
Objective: To investigate the effect of hsa_circ_0000392 (circ_0000392) on the radiosensitivity of cervical cancer cells and explore its potential mechanism. Methods: Cervical cancer tissues and adjacent normal tissues of 42 patients with cervical cancer who were confirmed pathologically for the first time in Huaihe Hospital of Henan University from 2016 to 2019 were collected. According to the patients' response to radiotherapy, the cancer tissues were divided into radio-sensitive tissues and radio-resistant tissues. The expressions of circ_0000392, miR-145-5p, and CRKL in radiation-sensitive, radiation-resistant cervical cancer tissues and Hela, SiHa cells were detected by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and western blot. SiRNA circ_0000392, miR-145-5p mimic, miR-145-5p inhibitor, pcDNA 3.1-CRKL and its negative control were transfected into HeLa and Siha cells, respectively. After radiation induction, the survival fraction of cells was detected by clone formation assay, apoptosis was detected by flow cytometry, and the expressions of apoptosis-related proteins Bax and Bcl-2 and ERK pathway protein p-ERK1/2 and ERK1/2 were detected by western blot. The targeting relationship between circ_0000392, miR-145-5p and CRKL was verified by dual luciferase reporter gene assay. The effect of circ_0000392 on radiotherapy sensitivity of cervical cancer in vivo was observed in the tumor formation experiment in nude mice. Results: circ_0000392 and CRKL were upregulated in radiation-resistant tissues and cancer cells of cervical cancer, while miR-145-5p was downregulated. The clone formation numbers of Hela and SiHa cells in si-circ_0000392#1+ 6 Gy group were (78.67±10.97) and (71.00±9.54), respectively, which were lower than those in si-Ctrl+ 6 Gy group [(176.00±22.27) and (158.33±17.56), respectively]. The apoptosis rates were (41.55±3.40)% and (31.41±3.29)%, respectively, which were higher than those in si-Ctrl+ 6 Gy group [(15.91±1.37)% and (13.70±1.89)%, P<0.05]. The protein expression of Bax was higher than that of si-Ctrl+ 6 Gy group, and the protein expressions of Bcl2 was lower than those of si-Ctrl+ 6 Gy group. The clone formation numbers of Hela and SiHa cells in si-circ_0000392#1+ miR-145-5p inhibitor+ 6 Gy group were (171.33±25.01) and (137.00±21.66), higher than those in si-circ_0000392#1+ inhibitor NC+ 6 Gy group [(84.67±17.79) vs (71.00±11.00), P<0.05]. The apoptosis rates were (17.41±2.58) % and (15.96±1.25) %, lower than those of si-circ_0000392 #1+ inhibitor NC+ 6 Gy [(40.29±2.92)% and (30.82±2.34)%, respectively, P<0.05]. The expression of Bax protein was lower than that of si-circ_0000392#1+ inhibitor NC+ 6 Gy group, and the expressions of Bcl2 protein were higher than those of si-circ_0000392#1+ inhibitor NC+ 6 Gy group. Circ_0000392 can target miR-145-5p, and CRKL is the downstream target gene of miR-145-5p. The clone formation numbers of Hela and SiHa cells in miR-145-5p mimic+ 6 Gy group were (74.33±10.02) and (66.00±12.17), respectively, which were lower than those of mimic NC+ 6 Gy group [(197.67±17.21) vs (157.67±11.59), respectively, P<0.05]. The apoptosis rates were (45.58±2.16)% and (32.10±3.55)%, higher than those of mimic NC+ 6 Gy group [(15.85±2.45)% and (13.99±1.69)%, respectively, P<0.05]. The expression of Bax protein was higher than that of the mimic NC+ 6 Gy mimic group, and the expression of Bcl2 protein was lower than that of the mimic NC+ 6 Gy group. The clone formation numbers of Hela and SiHa cells in miR-145-5p mimic+ pcDNA-CRKL+ 6 Gy group were (158.00±15.88) and (122.33±13.65), respectively, which were higher than those of miR-145-5p mimic+ pcDNA+ 6 Gy group [(71.33±8.02) vs (65.67±12.22), P<0.05]. The apoptosis rates were (19.50±3.45)% and (17.04±0.94)%, respectively, which were lower than those of miR-145-5p mimic+ pcDNA+ 6 Gy group [(44.33±2.36)% and (32.05±2.76)%, respectively, P<0.05]. The expression of Bax protein was lower than that of miR-145-5p mimic+ pcDNA group+ 6 Gy group, and the expression of Bcl2 protein was higher than that of miR-145-5p mimic+ pcDNA+ 6 Gy group. Sh-circ_0000392 group had smaller tumor volume and decreased tumor weight (P<0.05). The relative mRNA expression levels of circ_0000392, miR-145-5p and CRKL and the relative protein expression levels of CRKL, Bcl-2 and p-ERK1/2 were decreased, while the relative expression level of Bax protein was increased (P<0.05). Conclusion: Circ_0000392 could enhance the radiosensitivity of cervical cancer cells, and its mechanism may be related to the regulation of CRKL/ERK signaling pathway by targeting miR-145-5p, which provides a new reference for enhancing the radiosensitivity of cervical cancer cells.
Subject(s)
Animals , Mice , Female , Humans , Uterine Cervical Neoplasms/radiotherapy , bcl-2-Associated X Protein/genetics , Mice, Nude , Proto-Oncogene Proteins c-bcl-2/genetics , Apoptosis , MicroRNAs/genetics , Cell Proliferation , Cell Line, TumorABSTRACT
BACKGROUND/OBJECTIVES@#Tibetan tea is a kind of dark tea, due to the inherent complexity of natural products, the chemical composition and beneficial effects of Tibetan tea are not fully understood. The objective of this study was to unravel the composition of Tibetan tea using knowledge-guided multilayer network (KGMN) techniques and explore its potential antioxidant and hypolipidemic mechanisms in mice.MATERIALS/METHODS: The C57BL/6J mice were continuously gavaged with Tibetan tea extract (T group), green tea extract (G group) and ddH 2 O (H group) for 15 days. The activity of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in mice was detected.Transcriptome sequencing technology was used to investigate the molecular mechanisms underlying the antioxidant and lipid-lowering effects of Tibetan tea in mice. Furthermore, the expression levels of liver antioxidant and lipid metabolism related genes in various groups were detected by the real-time quantitative polymerase chain reaction (qPCR) method. @*RESULTS@#The results showed that a total of 42 flavonoids are provisionally annotated in Tibetan tea using KGMN strategies. Tibetan tea significantly reduced body weight gain and increased T-AOC and SOD activities in mice compared with the H group. Based on the results of transcriptome and qPCR, it was confirmed that Tibetan tea could play a key role in antioxidant and lipid lowering by regulating oxidative stress and lipid metabolism related pathways such as insulin resistance, P53 signaling pathway, insulin signaling pathway, fatty acid elongation and fatty acid metabolism. @*CONCLUSIONS@#This study was the first to use computational tools to deeply explore the composition of Tibetan tea and revealed its potential antioxidant and hypolipidemic mechanisms, and it provides new insights into the composition and bioactivity of Tibetan tea.
ABSTRACT
In recent years， the continuous advances in material sciences and techniques have helped with the establishment and development of liver organoids that can simulate the structure and function of organs in vivo. In addition to the research on traditional biological factors， the construction of microenvironments with different mechanical cues to investigate the influence of mechanical stimulation on the growth of liver organoids has also become a research focus. This article first discusses the development of liver organoids and then reviews the influence of mechanical forces of different properties on the formation of liver organoids， so as to lay a foundation for the construction of more complex and ordered liver organoids in vitro and provide ideal research models for understanding the interaction between biological and mechanical factors in the formation of liver organoids.
ABSTRACT
【Objective】 To explore the relevant risk factors of Henoch-Schonlein purpura (HSP) recurrence so as to provide some theoretical basis for early identification of children prone to recurrence. 【Methods】 The clinical data of 417 children with HSP hospitalized in Department of Pediatrics, The First Affiliated Hospital of Xi’an Jiaotong University, in the past five years were collected and followed up. They were divided into recurrent group and non-recurrent group. Cox regression analysis was used for univariate and multivariate analysis, and finally the independent risk factors for HSP recurrence were screened. 【Results】 A total of 417 children with initial onset of HSP were included in the study. During the follow-up period of 14 to 60 months, 78 cases recurred, and the recurrence rate was 18.7%. 94.9% of the children had relapse within 1 year. The results of univariate Cox regression analysis showed that age >7 years old at the time of onset, history of infection, history of strenuous exercise, duration of rashes more than 4 weeks, high level of neutrophil-to-lymphocyte ratio (NLR), and high level of platelet-to-lymphocyte ratio (PLR) were all risk factors for HSP recurrence (P7 years old at the time of onset, history of infection, history of strenuous exercise, duration of rashes for more than 4 weeks at the first onset, and high PLR level were independent risk factors for HSP recurrence (P 7 years at the time of onset, with a history of infection, vigorous exercise, rashes lasting more than 4 weeks, and high PLR level, nursing should be strengthened after discharge to avoid infection and vigorous exercise and increase the frequency of follow-up.
ABSTRACT
【Objective】 To explore the efficacy of flexible ureteroscopic lithotripsy (FURL) under local anesthesia (LA) in the treatment of upper urinary tract calculi, and to analyze the risk factors of pain. 【Methods】 Clinical data of 255 patients treated during Apr.2022 and Sep.2022 were reviewed, including visual analogue score (VAS) of pain during ureteroscopy, sheath placement, holmium laser lithotripsy, 1 hour and 8 hours after operation. VAS ≥5 was defined as significant pain. Clinical and follow-up data of the significant pain group and non-significant pain group were analyzed with logistic regression to analyze the risk factors of pain in FURL under LA. 【Results】 Altogether 198 patients (77.6%) successfully completed the operation, and the stone-free rate (SFR) was 89.9% (178/198). The VAS of ureteroscopy was the highest (4.49±1.08), and 73 patients (28.6%) experienced significant pain. Univariate analysis showed that significant pain was associated with gender, previous surgical history, age, body mass index (BMI), education level, and ASA classification (P<0.05). Multivariate analysis showed that male (OR=2.896, 95%CI:1.413-5.933, P=0.040) and BMI≥28 (OR=7.776, 95%CI:2.268-26.657, P=0.001) were independent risk factors of significant pain, while age ≥65 years (OR=0.237, 95%CI:0.083-0.672, P=0.007) and previous surgical history (OR=0.156, 95%CI:0.032-0.754, P=0.021) were the protective factors. 【Conclusion】 It is feasible and effective to treat upper urinary tract calculi with FURL under LA. The presence of significant pain is associated with factors such as gender, age, BMI and previous surgical history.
ABSTRACT
【Objective】 To compare the clinical efficacy of double and triple D-J ureteral stents in the treatment of benign secondary ureteral stricture. 【Methods】 Clinical data of 58 patients with benign secondary ureteral stricture treated at our hospital during Dec.2018 and Aug.2021 were retrospectively analyzed. According to the number of D-J ureteral stents, the patients were divided into the double group (n=29) and triple group (n=29). The clinical efficacy, complications during catheterization, and fluctuations of serum creatinine and hydronephrosis volume were compared and analyzed between the two groups. 【Results】 There were no significant differences in the general data, catheter duration and follow-up between the two groups (P>0.05). The triple group had more reduction of hydronephrosis than the double group [-22.8(32) cm3vs. -7.4(12) cm3, P=0.001] . There were no significant differences in the change of serum creatinine [-8.0(15)μmol·L-1 vs. -4.0(15) μmol·L-1, P =0.657] and incidence of complications (34.4% vs. 41.4%, P=0.588) between the two groups. The triple group had higher total effective rate than the double group (93.1% vs. 86.2%), but the difference was not statistically significant (P=0.666). There was no significant difference in the efficacy of balloon dilatation between the two groups (100.0% vs. 90.9%, P=0.407). 【Conclusion】 Both double and triple D-J ureteral stents are safe and effective in the treatment of benign secondary ureteral stenosis, but three D-J ureteral stents can better reduce hydronephrosis. Clinicians can choose appropriate indwelling scheme according to patients’ condition.
ABSTRACT
Objective: To investigate the differences of immune microenvironment between stage T1N3 and stage T3N0 breast cancer patients and explore the relationship between M1 macrophage infiltration and lymph node metastasis in breast cancer. Methods: Clinical information and RNA-sequencing (RNA-Seq) expression data of stage T1N3 (n=9) and stage T3N0 (n=11) breast cancer patients were extracted from Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) databases. Using CIBERSORT, the proportions of 22 types of immune cells were calculated, and then the differences of immune cell infiltration between stage T1N3 and T3N0 patients were compared. From 2011 to 2022, pathologic specimens were collected from breast cancer patients who underwent curative resection at the Cancer Hospital, Chinese Academy of Medical Sciences, including 77 at stage T1N3 and 58 at stage T3N0.The METABRIC database analysis results were verified by examining the density of M1 macrophages in tissues using dual-staining immunohistochemistry. Results: METABRIC data analysis showed M1 macrophage was the highest proportion, 15.85% in stage T1N3 breast cancer; M2 macrophage was the highest proportion, 13.07% in stage T3N0 breast cancer.M1 macrophage proportions were statistically different between patients with stage T1N3 and stage T3N0 (P=0.010). The dual-staining immunohistochemistry analysis of breast cancer tissues showed M1 macrophage density (median) of 62.0 and 38.0 cells/mm(2) for stage T1N3 and T3N0, respectively. The difference was statistically significant (P=0.002). Conclusion: The density of M1 macrophages is notably higher in stage T1N3 patients and is associated with lymph node metastasis.