In the field of biomedicine,two-dimensional(2D)cell lines and animal models have played an im-portant role in the study of cell pathways and drug targets.However,due to species differences between humans and other animals,and the lack of hierarchy,cellular diversity,and cell-cell or cell-matrix interactions,2D cell lines could not ful-ly reflect what cells actually look like in the human body.Organoids are three-dimensional(3D)in vitro culture models de-rived from autologous tissue stem cells,which make up for the defects of 2D culture and can simulate the structure and function of real human organs to a certain extent,providing new ideas for disease diagnosis and treatment.Among them,lung organoids(LO)are a typical case studying the development process of human lung and the generation principle of lung diseases.Relevant studies have provided help for the treatment of pulmonary fibrosis,lung cancer,lung injury and other diseases.This paper aims to summarize and analyze the research progress of lung organoids in recent years,and fur-ther summarize the application of LO in the diagnosis and treatment of lung diseases.

ObjectiveTo study the effect of Tanreqing injection （TRQ） on the pulmonary flora in the rat model of chronic obstructive pulmonary disease （COPD）. MethodWistar rats were randomized into control， model， and TRQ groups. The rats in other groups except the control group were treated by smoking combined with intratracheal instillation of lipopolysaccharide for the modeling of COPD. The TRQ group was intraperitoneally injected with TRQ （2 g·kg^-1）. At the end of the experiment， after blood collection from the abdominal aorta of the rats， the lung tissue was collected for hematoxylin-eosin and picric sirius red staining to reveal the pathological changes. The lung lavage fluid was collected， and the diversity and relative abundance of lung flora in different groups were analyzed by 16S rDNA amplicon sequencing. ResultThe lungs of the control group were normal， and those of the model group showed neutrophil infiltration， telangiectasia， lung hemorrhage and emphysema in individual cases， and thickening of collagen fibers in the trachea. Compared with the model group， the TRQ group showed significantly improved lungs and recovered collagen fibers. The MLI analysis showed that compared with the control group， the model group showcased increased alveolar space （P<0.01）， which was reduced in the TRQ group （P<0.05）. Compared with the control group， the model group showed increased wall thickness （P<0.01）， and the increase was attenuated in the TRQ group （P<0.01）. TRQ increased the Simpson index and altered the α diversity of pulmonary flora. The results of principal co-ordinate analysis showed that TRQ changed the β diversity and reduced the β diversity index of pulmonary flora. At the genus level， the model group showed increased relative abundance of g_Bacillus and g_Brevundimonas and decreased relative abundance of g_Pseudomonas， compared with the control group. After treatment with TRQ， the relative abundance of g_Stenotrophomonas increased， and that of g_Bacillus decreased. The LEfSe of differential taxa between groups showed that the modeling increased the relative abundance of g_Lachnospiraceae_NK4A136_group， and TRQ treatment increased the relative abundance of g_Rhodococcus and g_Stenotrophomonas. ConclusionTRQ can regulate the diversity of pulmonary flora and restore the balance of bacterial genera in the rat model of COPD， which may be one of the mechanisms of the prevention and treatment of COPD with TRQ.

Objective:Migraine and tremor share some genetic mutation sites,and clinical studies have also confirmed their correlation.This study aims to explore the clinical and electrophysiological characteristics of migraine patients with concomitant tremor,and to analyze the relevant influential factors of tremor occurrence. Methods:We retrospectively analyzed the clinical data of 217 migraine patients who visited the Third Affiliated Hospital of Qiqihar Medical University from June 2022 to October 2023.The Clinical Rating Scale for Tremor(CRST),Numerical Rating Scale(NRS),Generalized Anxiety Disorder-7(GAD-7),and Patient Health Questionnaire-9(PHQ-9)were respectively used to assess the tremor symptoms,degree of headache,anxiety,and depression of patients.All patients underwent routine head MR scanning and electromyography examination,and were divided into a migraine with tremor group and a migraine without tremor group based on the electromyogram examination. Results:The migraine with tremor group and the migraine without tremor group were included 52 patients(23.96%)and 165 patients(76.04%),respectively.Compared with the migraine without tremor group,the migraine with tremor group had a longer course and duration of headache,higher frequency of headache attacks,higher NRS score,GAD-7 score,and PHQ-9 score,and fewer weekly physical exercises.The differences were statistically significant(all P<0.05).There were no statistically significant differences in the presence or absence of prodromal headache and white matter hyperintensities(WMHs)between the 2 groups(both P>0.05).The evaluation results of the CRST showed that out of 217 migraine patients,39 patients(17.97%)were accompanied by tremors.The electromyographic results showed that all 52 migraine patients with tremors had upper limb tremors,including 28 migraine patients with postural tremors and 24 migraine patients with static tremors.Compared with the migraine patients with static tremors,the migraine patients with postural tremors had lower average frequency,peak frequency,and headache onset frequency(all P<0.05).Multiple linear regression analysis showed that frequency of physical exercise,duration of illness,frequency of headache attacks,NRS score,GAD-7 score,and PHQ-9 score were risk factors for migraine patients with concomitant tremors(all P<0.05). Conclusion:Patients with migraine mainly experience upper limb postural tremors.Reduced physical exercise,long course of disease,long duration of headache,frequent headache attacks,severe headache,anxiety,and depression are risk factors for migraine patients with concomitant tremors.

Cardiotoxicity is a serious complication of antineoplastic drugs. With the continuous development of antineoplastic drugs, immune checkpoint inhibitors have been used in the treatment of a variety of cancers. PD-1/PD-L1 immune checkpoint inhibitors have been widely concerned in recent years. This article reviews the manifestations and possible mechanisms of cardiotoxicity induced by PD-1/PD-L1 immune checkpoint inhibitors, and the detection methods and treatment of cardiotoxicity.

Objective To discuss the correlation between gastrointestinal symptoms concomitant with chronic migraine and sleep disorders,anxiety and depression.Methods Self-rating Anxiety Scale (SAS),Self-rating Depression Scale (SDS),Pittsburgh Sleep Quality Index (PSQI) and Gastrointestinal Symptom Rating Scale (GSRS) were adopted to conduct comparative analysis on gastrointestinal symptoms,anxiety,depression,sleep disorders,and related risk factors in 151 chronic migraine patients who conformed to International Headache Society International Classification of Headache Diseases-Ⅱ diagnostic code and 151 healthy controls in the Third Affiliated Hospital of Qiqihar Medical University from January 2015 to December 2017.Results PSQI scores (8.46±2.54 vs 4.53±1.95,t=2.913,P＜0.05),SAS scores (53.24± 11.66 vs 39.58±8.63,t=3.112,P＜0.05),SDS scores (51.09±8.80 vs 36.11±5.74,t=3.520,P＜0.05),and GSRS scores (4.53 (3.74,5.32) vs 1.29 (0.73,1.31),Z=30.804,P＜0.05) were all higher in migraine patients than that in healthy controls,and the differences were all statistically significant.In chronic migraine patients,139 (92.05％) had one gastrointestinal symptom at least,and the overall incidence of each gastrointestinal symptom was abdominal distension (60 cases,39.74％),exhaust increase (51 cases,33.77％),and endless defecation (43 cases,28.48％),etc.,in turn.Multiple regression analysis showed that headache degree,frequency,PSQI scores,SAS scores,and SDS scores were the main risk factors of gastrointestinal symptoms concomitant with chronic migraine.GSRS scores of chronic migraine patients presented positive correlation with PSQI scores (r=0.65,P=0.000),SAS scores (r=0.42,P=0.000),and SDS scores (r=0.48,P=0.000),respectively.Conclusion Chronic migraine patients are easy to be accompanied with gastrointestinal symptoms,and headache degree and frequency,sleep disorders,depression and anxiety are the main risk factors of concomitant gastrointestinal symptoms.

Objective To investigate the potential effects and mechanism on peroxisome proliferator-activated receptor γ-toll-like receptor 4-tumor necrosis factor-α (PPARγ-TLR4-TNF-α) targeted pathway on hyperglycemia induced myocardium inflammation and oxidative stress. Methods Thirty-two Japanese healthy adult rabbits were randomly divided into four groups with 8 rabbits in each group: normal control group (NC group), diabetes mellitus group (DM group), diabetes mellitus + pioglitazone 4 mg·kg-1·d-1 and 8 mg·kg-1·d-1 groups (DM+PGZ 4 mg and 8 mg groups). DM model was reproduced by alloxan of 150 mg/kg through auricular vein injection. On the same day of successful DM model reproduction, the diabetic rabbits were fed with corresponding dose of pioglitazone in DM+PGZ 4 mg and 8 mg groups, but the rabbits in NC group were not challenged. After 8 weeks of feeding, venous blood of left jugular vein bifurcation and myocardium tissue were harvested respectively for the determination of inflammation and oxidative stress parameters. TNF-α, interleukin-1 (IL-1), adiponectin (ADP), nitric oxide (NO) and total nitric oxide synthase (NOS) levels were determined by enzyme linked immunosorbent assay (ELISA), myeloperoxidase (MPO) activity was determined by colorimetric method, superoxide dismutase (SOD) activity was determined by hydroxylamine method, malondialdehyde (MDA) was determined by thiobarbituric acid colorimetric method, and catalase (CAT) activity was determined by UV spectrophotometry method. In addition, the mRNA expressions of TNF-α and TLR4 were determined by real-time quantitate reverse transcription-polymerase chain reaction (RT-qPCR). Results ① IL-1 and TNF-α in serum and myocardium of model rabbits were significantly increased, ADP was significantly decreased, and the mRNA expressions of TNF-α and TLR4 in myocardium were significantly increased, indicating a significant inflammatory reaction. The inflammatory reaction in pioglitazone intervention groups was significantly reduced, TNF-αand IL-1 levels in serum and myocardium of DM+PGZ 4 mg and 8 mg groups were significantly decreased as compared with those of DM group [serum: TNF-α(ng/L) was 268.33±46.57, 261.34±33.73 vs. 331.40±69.05, myocardium: TNF-α (ng/L) was 144.72±26.90, 139.59±14.59 vs. 177.48±27.40; serum: IL-1 (ng/L) was 24.40±2.56, 23.35±3.13 vs. 30.08±5.44, myocardium: IL-1 (ng/L) was 21.26±2.85, 20.54±2.75 vs. 24.78±3.60, all P < 0.05], and ADP levels were significantly increased [serum (μg/L): 19.64±8.85, 20.54±7.47 vs. 15.45±3.06, myocardium (μg/L): 10.31±2.22, 11.49±3.42 vs. 7.76±1.77, all P < 0.05], and the mRNA expressions of TNF-α and TLR4 in myocardium were significantly decreased (TNF-αmRNA: 0.15±0.05, 0.14±0.06 vs. 0.25±0.09; TLR4 mRNA: 0.57±0.17, 0.40±0.18 vs. 0.75±0.35, all P < 0.05). ②Oxidative stress in serum and myocardium of model rabbits was significantly increased, SOD, NO, and total NOS levels were significantly decreased while the serum CAT and MDA levels were significantly increased without effect on MPO. Compared with the DM group, SOD and NO levels in serum and myocardium were significantly increased in DM+PGZ 4 mg and 8 mg groups [serum: SOD (U/L) was 571.39±40.85, 609.28±54.47 vs. 535.10±37.08, myocardium:SOD (U/mg) was 55.74±8.12, 53.60±9.87 vs. 42.26±12.34; serum: NO (μmol/L) was 2.95±0.51, 2.99±0.43 vs. 2.03±0.78, myocardium: NO (nmol/mg) was 1.95±0.37, 2.11±0.26 vs. 1.56±0.33, all P < 0.05], the serum MDA levels were significantly decreased (μmol/L: 20.11±2.34, 19.70±2.02 vs. 23.07±3.06, both P < 0.05), while no significant effect on CAT. There was no significant difference in parameter of inflammatory and oxidative stress between the two pioglitazone intervention groups. Conclusion 4 mg·kg-1·d-1 pioglitazone could activate PPARγ-TLR4-TNF-α targeted pathway, thus inhibit inflammatory and oxidative stress factors expression, and down-regulate hyperglycemia induced myocardium inflammatory and oxidative stress level, but the effect did not show a dose dependent manner.

Objective To investigate changes in the number and function of endothelial progenitor cells (EPCs) in elderly patients with permanent atrial fibrillation(AF).Methods This prospective study included 35 elderly patients in each the permanent atrial fibrillation group and the control group.The numbers of circulating CD34+/KDR+ cells in the two groups were determined by flow cytometry.After two sets of peripheral blood samples were taken,mononuclear cells were isolated through density gradient centrifugation and cultured in vitro.EPC colonies were identified by the methylthiazolyldipheny-tetrazolium(MTT) assay and adhesion assay.The proliferation,adhesion and vasculogenesis of EPC colonies were determined by Matrigel culture.Enzyme-linked immunosorbent assay and nitric acid reductase assay were used to measure the secretion of vascular endothelial growth factor (VEGF) and nitric oxide (NO) in EPCs.Results The numbers of CD34+/KDR+ cells were lower in the AF group than in the control group (20.0±12.7)/104 vs.(77.9±58.9)/104 (P＜0.05).The number of EPC colonies in the atrial fibrillation group was significantly lower than that in the control group (1.8 ± 0.6) CFU vs.(3.5 ± 0.8) CFU (P ＜ 0.01).The proliferation,adhesion and vasculogenesis of EPC colonies in the AF group decreased,compared with the control group (each P＜0.01 or 0.05).Paracrine secretion of VEGF in the AF group (27.4±9.9)ng/L was lower than that in the control group (41.9±7.3)ng/L (P＜0.01) and paracrine production of NO in the AF group also decreased (P＜0.05).Conclusions EPCs In elderly patients with permanent atrial fibrillation show decreased numbers and reduced proliferation,adhesion and vasculogenesis.Paracrine VEGF and NO secretion is down as well.

Objective To explore the effect of the temperature-sensitive hydrogel and platelet-rich plasma(PRP)complex on the healing of anterior cruciate ligament (ACL)partial tear in a rat model.Method PRP was processed according to an established method and then mixed with poly(ethylene glycol)monomethyl ether and poly(lactic-co-glycolic acid)(mPEG-PLGA).A total of 110 right knees of Sprague-Dawley male rats (12-week old)were included and randomly divided into a healthy control group (n=10),a lesion control group(n=60)and a study group(n=40).The saline and mPEG-PLGA-PRP complex was applied at the lesion site respectively.Samples were harvested 0,2 weeks and 6 weeks post-operatively and the histological and biomechanical changes were observed and compared among the 3 groups.Results Histologically,at 6 weeks after surgery,the torn ACL of the study group had been partially healed,with decreased number of inflammatory cells and new fibrous tissues and micro-vessels appearing.The ligament maturity index revealed a significantly higher score in the study group than the lesion control group(20.6 ± 4.9 vs.4.7 ± 1.0,P＜0.01).Biomechanically,at 6 weeks after surgery,the tensile strength of the study group was significantly higher than the contro] groups(52.7 ± 11.2 vs.30.3 ± 8.8,P＜0.05).Conclusion At six weeks after surgery,the mPEG-PLGA-PRP complex can enhance the healing of ACL partial tear,ahhough the ligament was not recovered to the norma] state.

Objective The purpose of this study was to detect the degradation degree of long-term formalin fixed tissue and to compare the detection rate of STR with SNP. Methods DNA was extracted from 24 formalin-fixed tissues stored at -20 ℃ for five years, and the concentration and degradation index of DNA was quantified with Quantifiler? Trio DNA Kit. A 55-SNP multiplex SNaPshot assay and PowerPlex? 21 system were used to amplify SNP and STR loci, respectively. Results The results showed that the degradation indexes of 24 specimens were ranged from 1~8. The SNP genotypes of the 24 specimens were completely consistent with the non-degraded DNA from the same individuals and the successful genotyping rate was 100%. However, 33 allele dropouts were observed with STR genotyping in 8 samples, of which the degradation index was higher than 2.6, and the fragment size of the 75.8% allele was longer than 300bp. The likelihood ratio based on 16 typable STR loci in the sample was close to that onthe basis of 54 SNPs. However, likelihood ratio based on more than 17 STR loci was over that accord to 54 SNPs. There was a negative correlation between the fragment size of STR and the allele detection rate, and a negative correlation also observed between the degradation index of samples and the allele detection rate except for two samples with mild degradation. Conclusion This study validated that the long-term formalin-fixed tissues were susceptible to degradation, and the SNP was more suitable for detecting these tissues than STR typing system. However more SNP loci are needed to test in order to increase the discrimination power.

Objective To explore the mutation types and disciplines of STR commonly used in forensic in gynecologic and breast cancerand investigate the application of microdissection in forensic practice involving tumor tissue. Methods DNA of tumor tissues, adjacent normal tissues and peripheral blood from 62 patients with breast cancer, 62 patients with gynecologic cancer and 10 patients with benign gynecologic tumor were amplified by PowerPlex 21 System kit and Argus X-12 kit. Capillary electrophoresis of PCR products was carried out on an ABI 3130 Genetic Analyzer to obtain genotypes. Some tumor tissues with STR variation were microdissected. Results The genotype of peripheral blood in cancer patient was consistent with that of corresponding normal tissue. 4 types of STR variations were found in 46.77% gynecologic cancer tissues, compared with that in benign tumor tissues and breast cancer, the difference of STR variation was significant(P<0.01,P=0.009). The genotype of stromal cells separated by microdissection was consistent with that of corresponding adjacent normal tissue. Conclusion The STR loci detected in the study with poor stability are not suitable for forensic cases involving gynecologic cancer tissues. The genotype of stromal cells separated accurately from tumor tissues by microdissection could represent the normal DNA genotype of the individual with cancer. Microdissection is an effective solution in forensic cases with tumor tissues.