Protective effect of Qingjie Huagong decoction on pancreatic tissue of mice with severe acute pancreatitis by regulating the NOD-like receptor protein 3/Toll-like receptor 4/nuclear factor-kappa B signaling pathway / 临床肝胆病杂志

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.

Research Progress of Traditional Chinese Medicine Intervention in Nrf2/HO-1 Signaling Pathway for Acute Pancreatitis： A Review / 中国实验方剂学杂志

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.

Study on the feasibility of the application of organ dose modulation technique of CT scanning for the eye / 中国医学装备

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.

Estimation of genotoxicity threshold induced by acute exposure to neodymium nitrate in mice using benchmark dose / 环境与职业医学

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.

Comparative study on the clinical efficacy of intermittent oral administration of fosfomycin trometamol for a long time after removal of upper urinary infection stones / 重庆医学

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.

Application and research progress of fecal microbiota transplantation in refractory diarrhea / 解放军医学杂志

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.

Codonopsis polysaccharide protected LPS-induced acute lung injury by inhibiting MAPK/NF-κB signaling pathway in mice / 实用医学杂志

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.

Application research of PGT in blocking the inheritance of novel mutations in the PKHD1 gene in autoso-mal recessive polycystic kidney disease pedigrees / 实用医学杂志

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.

Implantation of Adipose-Derived Mesenchymal Stromal Cells (ADSCs)-Lining Prosthetic Graft Promotes Vascular Regeneration in Monkeys and Pigs

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.

Two lumbar fusion regimens in treatment of single-level lumbar degenerative diseases based on propensity score matching / 中国组织工程研究

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.

Effect of carbonic anhydrase 9 on hypoxia-induced proliferation of retinal microvascular endothelial cells in preterm fetus / 中国新生儿科杂志

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.

Prevention and management of VKC in China from the Asian VKC expert consensus / 中华实验眼科杂志

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.

Clinical analysis of P2Y2 receptor agonist in the treatment of diabetic dry eye / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To evaluate the clinical efficacy of P2Y2 agonist diquafosol sodium(DQS)eye drops in the treatment of diabetic dry eye.METHODS: A total of 80 patients(160 eyes)with diabetic dry eye who admitted to our hospital from January 2022 to March 2022 were selected. They were randomly divided into study group and control group. A total of 40 patients(80 eyes)in the study group were treated with 3% DQS eye drops and 40 patients(80 eyes)in the control group were treated with 0.3% sodium hyaluronate eye drops. The ocular surface disease index(OSDI)score, non-invasive tear meniscus height(NITMH), first non-invasive tear film break-up time(NIBUTf), average non-invasive tear film break-up time(NIBUTav), tarsal gland loss score, lipid layer thickness grade and bulbar redness analysis(including conjunctival grade and ciliary grade), were examined before treatment and at 1wk, 1 and 3mo after treatment, respectively. Furthermore, corneal fluorescence staining and conjunctival lissamine green staining were analyzed based on the ocular surface staining score(OSS), and the conjunctival impression cytology and confocal microscopy were evaluated before and 3mo after treatment, respectively.RESULTS: There were no differences in OSDI score, tarsal gland loss score, conjunctival grade score and ciliary grade score between the two groups before and after treatment(P&#x0026;#x003E;0.05). OSS scores in the study group were lower than those in the control group, while NITMH, NIBUTf and NIBUTav were higher than those in the control group at 1 and 3mo after treatment(P&#x0026;#x003C;0.05). After 3mo of treatment, the density of conjunctival goblet cells increased and corneal dendritic cells decreased in the study group compared with the baseline(all P&#x0026;#x003C;0.05), while there were no significant changes in the control group compared with the baseline(all P&#x0026;#x003E;0.05).CONCLUSION: 3% DQS eye drops were effective in treating diabetic dry eye without serious complications.

Preparation of chitosan hydrochloride stabilized emulsion and its immunostimulatory effect / 生物工程学报

In order to increase the ability of oil-emulsion adjuvant to stimulate cellular immunity, chitosan hydrochloride with positive charge was selected to stabilize oil-in-water emulsion (CHE). In this paper, model antigen ovalbumin was selected to prepare vaccines with emulsion adjuvant, commercial adjuvant or no adjuvant. The emulsion was characterized by measuring the particle size, electric potential and antigen adsorption rate. BALB/c mice were immunized by intramuscular injection. Serum antibody levels, the numbers of IL-4-secreting cells in splenocytes, cytotoxic T lymphocyte (CTL) response, and the expression of central memory T cells were measured to evaluate the immunostimulatory effect. The results showed that chitosan hydrochloride can effectively stabilize the emulsion. The emulsion size is about 600 nm, and the antigen adsorption rate is more than 90%. After immunization, CHE could increase serum antibodies levels and increase IL-4 secretion. Expression of CTL surface activation molecules was also increased to stimulate CTL response further and to increase the CD44+CD62L+ in T cells proportion. CHE as adjuvant can stimulate humoral and cellular immunity more efficiently, and is expected to extend the duration of protection.

Construction of a Prognostic Model of Multiple Myeloma Based on Metabolism-Related Genes / 中国实验血液学杂志

OBJECTIVE@#To screen the prognostic biomarkers of metabolic genes in patients with multiple myeloma (MM), and construct a prognostic model of metabolic genes.@*METHODS@#The histological database related to MM patients was searched. Data from MM patients and healthy controls with complete clinical information were selected for analysis.The second generation sequencing data and clinical information of bone marrow tissue of MM patients and healthy controls were collected from human protein atlas (HPA) and multiple myeloma research foundation (MMRF) databases. The gene set of metabolism-related pathways was extracted from Molecular Signatures Database (MSigDB) by Perl language. The biomarkers related to MM metabolism were screened by difference analysis, univariate Cox risk regression analysis and LASSO regression analysis, and the risk prognostic model and Nomogram were constructed. Risk curve and survival curve were used to verify the grouping effect of the model. Gene set enrichment analysis (GSEA) was used to study the difference of biological pathway enrichment between high risk group and low risk group. Multivariate Cox risk regression analysis was used to verify the independent prognostic ability of risk score.@*RESULTS@#A total of 8 mRNAs which were significantly related to the survival and prognosis of MM patients were obtained (P<0.01). As molecular markers, MM patients could be divided into high-risk group and low-risk group. Survival curve and risk curve showed that the overall survival time of patients in the low-risk group was significantly better than that in the high risk group (P<0.001). GSEA results showed that signal pathways related to basic metabolism, cell differentiation and cell cycle were significantly enriched in the high-risk group, while ribosome and N polysaccharide biosynthesis signaling pathway were more enriched in the low-risk group. Multivariate Cox regression analysis showed that the risk score composed of the eight metabolism-related genes could be used as an independent risk factor for the prognosis of MM patients, and receiver operating characteristic curve (ROC) showed that the molecular signatures of metabolism-related genes had the best predictive effect.@*CONCLUSION@#Metabolism-related pathways play an important role in the pathogenesis and prognosis of patients with MM. The clinical significance of the risk assessment model for patients with MM constructed based on eight metabolism-related core genes needs to be confirmed by further clinical studies.

Diagnosis and Treatment of Klebsiella Pneumoniae Combined with Enterobacter Cloacae in Lumbar Intervertebral Space Infection / 中山大学学报(医学科学版)

ObjectiveTo explore the clinical diagnosis and treatment of rare primary lumbar intervertebral space infection with Klebsiella pneumoniae and Enterobacter cloacae， and provide clinical experience for the diagnosis and treatment of this rare spinal infection. MethodsAn elderly male patient with low back pain and numbness in the left lower extremity for more than 7 months， which aggravated for more than 1 week， was diagnosed with lumbar disc herniation after laboratory and imaging examinations. After admission， the symptoms became acutely aggravated， and re-examination of lumbar enhanced MRI showed local enhancement at the posterior edge of the L3/4 intervertebral space. The VAS score was 9 points， and the lumbar JOA score was 6 points. A posterior lumbar interbody fusion of L3-L5 was performed， and L3/4 intervertebral disc specimens were collected during the operation for bacterial culture. ResultsBacterial culture results showed Klebsiella pneumoniae and Enterobacter cloacae infection. The patient was treated with sensitive antibiotics for 6 weeks after the operation， and the patient was cured during the follow-up of half a year after the operation. ConclusionFor middle-aged and elderly patients with clinical manifestations of acute severe low back pain or lower extremity pain， the possibility of spinal infection should be considered when routine laboratory and imaging examinations suggest lumbar degenerative diseases.

Advances in live-imaging aging reporter mice / 生理学报

Aging is an independent risk factor for chronic diseases in the elderly, and understanding aging mechanisms is one of the keys to achieve early prevention and effective intervention for the diseases. Aging process is dynamic and systemic, making it difficult for mechanistic study. With recent advances in aging biomarkers and development of live-imaging technologies, more and more reporter mouse models have been generated, which can live monitor the aging process, and help investigate aging mechanisms at systemic level and develop intervention strategies. This review summarizes recent advances in live-imaging aging reporter mouse models based on widely used aging biomarkers (p16Ink4a, p21Waf1/Cip1, p53 and Glb1), and discusses their applications in aging research.

Muscone inhibits opening of mPTP to alleviate OGD/R-induced injury of HT22 cells / 中国中药杂志

This study aims to investigate the mechanism of muscone in inhibiting the opening of mitochondrial permeability transition pore(mPTP) to alleviate the oxygen and glucose deprivation/reoxygenation(OGD/R)-induced injury of mouse hippocampal neurons(HT22). An in vitro model of HT22 cells injured by OGD/R was established. CCK-8 assay was employed to examine the viability of HT22 cells, fluorescence microscopy to measure the mitochondrial membrane potential, the content of reactive oxygen species(ROS), and the opening of mPTP in HT22 cells. Enzyme-linked immunosorbent assay was employed to determine the level of ATP and the content of cytochrome C(Cyt C) in mitochondria of HT22 cells. Flow cytometry was employed to determine the Ca~(2+) content and apoptosis of HT22 cells. The expression of Bcl-2(B-cell lymphoma-2) and Bcl-2-associated X protein(Bax) was measured by Western blot. Molecular docking and Western blot were employed to examine the binding between muscone and methyl ethyl ketone(MEK) after pronase hydrolysis of HT22 cell proteins. After the HT22 cells were treated with U0126, an inhibitor of MEK, the expression levels of MEK, p-ERK, and CypD were measured by Western blot. The results showed that compared with the OGD/R model group, muscone significantly increased the viability, mitochondrial ATP activity, and mitochondrial membrane potential, lowered the levels of ROS, Cyt C, and Ca~(2+), and reduced mPTP opening to inhibit the apoptosis of HT22 cells. In addition, muscone up-regulated the expression of MEK, p-ERK, and down-regulated that of CypD. Molecular docking showed strong binding activity between muscone and MEK. In conclusion, muscone inhibits the opening of mPTP to inhibit apoptosis, thus exerting a protective effect on OGD/R-injured HT22 cells, which is associated with the activation of MEK/ERK/CypD signaling pathway.

Effects of Yinqi Sanhuang Jiedu Decoction （茵芪三黄解毒汤） on Liver Hardness and Epithelial Mesenchymal Transition in Hepatic Fibrosis Model Mice / 中医杂志

ObjectiveTo investigate the possible mechanism of compound Yinqi Sanhuang Jiedu Decoction （茵芪三黄解毒汤， YSJD） against the progression of hepatic fibrosis （HF）. MethodsThirty-eight C57BL/6J mice were randomly divided into blank group， model group， silybin group and low-， medium- and high-dose YSJD groups， with eight mice in the model group and six mice each in other groups. Except for the blank group， all mice were injected intraperitoneally with 10% carbon tetrachloride （CCl4） to establish a HF model， twice a week for 8 weeks. The drug intervention started one week after the first modeling； the low-， medium- and high-dose YSJD groups were given 8.325， 16.65 and 33.3 g/（kg·d） of YSJD suspension by gavage， respectively， while the silybin group was given 55 mg/（kg·d） of silybin suspension by gavage， and the blank group and the model group were given 0.2 ml normal saline by gavage， all for 8 weeks. The liver hardness of living mice was observed using a small animal ultrasound detector， and grey-scale ultrasound was recorded. The liver tissue was observed by Sirius scarlet staining， and the proportion of collagen fiber deposition was calculated. Liver function indicators including alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， and serum laminin （LN）， hyaluronic acid （HA）， pre-collagen type III （PCIII） and collagen type IV （CIV） were also detected. The protein expression of transforming growth factor β1 （TGF-β1）， Vimentin， E-cadherin， α smooth muscle actin （α-SMA） in liver tissue were detected. ResultsCompared to the blank group， the model group showed increased gray value， collagen deposition，serum ALT， AST， HA， LN and PCIII levels， decreased expression of E-cadherin， and increased expression of N-cadherin，α-SMA，Vimentin and TGF-β1 in liver tissues （P＜0.05）. Compared to the model group， the ultrasonic gray value and the proportion of collagen fiber deposition in liver of silybin group and YSJD medium- and high-dose groups decreased， and the serum ALT， AST， LN， HA and PCⅢ levels decreased. Compared to the model group， the expression of E-cadherin in liver tissues of silybin group and all three YSJD groups increased， while the expressions of N-cadherin， Vimentin， TGF-β1 and α-SMA decreased （P＜0.05）， and among them， most improvements were seen in the medium-dose YSJD group （P＜0.05）. ConclusionThe effect of YSJD on significantly reducing the extent of HF in mice caused by CCl4 may be related to its ability to regulate liver hardness and inhibit the occurrence of epithelial mesenchymal transition in mice.

Infiltration and significance of M1 macrophage in breast cancer: an analysis of METABRIC database and clinical validation / 中华肿瘤杂志

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.