BACKGROUND:Liver organoids are of great significance to elucidate the exact pathological mechanism of liver diseases and the treatment of liver diseases. OBJECTIVE:To summarize the basic research in this field at home and abroad,review the important research progress in the construction of liver organoids,disease modeling and transplantation therapy,and discuss the application prospect of combined tissue engineering technology of liver organoids. METHODS:The relevant articles included in PubMed and CNKI databases were searched.The English and Chinese search terms were"liver,organoids,liver diseases."The main search time was from April 2018 to April 2024.Duplicate literature was excluded by manual reading.Finally,94 articles were included for review and analysis. RESULTS AND CONCLUSION:The seed cells constructed by liver organoids are mainly concentrated in adult cells and pluripotent stem cells,which promote the generation of organoids by assisting various cytokines to participate in signal guidance and providing 3D microenvironment by extracellular matrix.However,the overall maturity is not high,which is expected to improve this problem by combining tissue engineering technology.In vitro disease modeling is mainly studied in the field of simple diseases and single-gene genetic diseases.Organoids highly retain patient genetic characteristics,and it is expected to simulate more complex liver diseases and clarify deeper pathological mechanisms by combining CRISPR-Cas9 gene correction and other emerging technologies.In vivo transplantation treatment,liver organoids can be safely and effectively implanted,showing amazing liver function replacement potential,tissue regeneration ability,and may also be combined with other tissue engineering materials to achieve therapeutic purposes.

Objective To investigate the effect of electromagnetic wave power density on the expression of glial fibrillary acidic protein(GFAP)in the hippocampus of SD rats under 1800 MHz electromagnetic wave irradiation,and whether it exhibits a"window effect".Methods Ninety-eight 4-week-old SPF-grade SD rats were randomly divided into 14 groups,with 7 rats in each group.Seven groups were exposed groups(frequency:1800 MHz,power densities:0.1 mW/cm2,0.3 mW/cm2,0.5 mW/cm2,0.7 mW/cm2,0.9 mW/cm2,1.0 mW/cm2,1.2 mW/cm2)and corresponding 7 groups were control groups(power density:0 mW/cm2).Exposure was conducted for 12 hours daily for 3 weeks.After exposure,Western Blot was used to detect the expression level of GFAP in the hippocampal tissue,and immunohistochemistry staining was performed to determine the average optical density(MOD)value of GFAP-positive expression products in the DG,CA3,and CA1 regions of the hippocampal tissue,to determine the power density window of GFAP expression in the hippocampus of SD rats under 1800 MHz exposure.Results At power densities of 0.1 mW/cm2 and 0.3 mW/cm2,Western Blot results showed increased expression of GFAP in the rat hippocampus(P<0.05),and immunohistochemistry staining demonstrated increased MOD values of GFAP in the three regions(P<0.05).Conclusion Long-term exposure to 1800 MHz elect-romagnetic radiation has a"window effect"on the expression of GFAP in the DG,CA3,and CA1 regions of the hippocampus in SD rats,with power density windows of 0.1 mW/cm2 and 0.3 mW/cm2.

Objective To observe the effects of electroacupuncture on intestinal flora and serum inflammatory factors in rabbit model with rheumatoid arthritis(RA);To explore the mechanism of its therapeutic effect on RA.Methods RA model rabbits were established by ovalbumin induction combined with Freund's complete adjuvant,and the rabbits after successful modeling were randomly divided into model group,probiotic group and electroacupuncture group,with 6 rabbits in each group.Another 6 rabbits were set as the normal group.The electroacupuncture group received electroacupuncture at the bilateral"Zusanli"and"Dubi"for 30 minutes,the probiotic group was given probiotic capsules solution(14.5 mg/kg)by gavage,once a day,for two consecutive weeks.The knee joint circumference and pain threshold of rabbits were measured,histopathological morphological changes of colonic tissue and synovial tissue ws observed by HE staining,16S rDNA sequencing was used to analyze structural changes of intestinal flora,the contents of TNF-α,IL-1β and IL-6 in serum were detected by ELISA.Results Compared with the normal group,the circumference of knee joint of rabbits in the model group increased significantly,the pain threshold was significantly decreased(P<0.01);the colonic mucosal damage was serious,the goblet cells were missing,a large number of inflammatory cells were infiltrate;the joint capsule synovial surface was rough,the synovial cell layer was hyperplasia and thickening,the synovial tissue inflammatory cell infiltration was obvious;the number and evenness of gut microbiota species decreased,while the relative abundance of Proteobacteria,Firmicutes,and Bacteroidetes decreased(P<0.05,P<0.01),the relative abundance of Desulfobacteria increased(P<0.01),while the relative abundance of Campylobacter,Lawsonella,and Pseudomonas increased(P<0.01),while the relative abundance of Heshanomonas and Herbaspirillum decreased(P<0.01);the contents of serum TNF-α,IL-1β and IL-6 increased(P<0.01).Compared with the model group,the knee joint circumference of the probiotic group and the electroacupuncture group decreased,the pain threshold increased(P<0.01,P<0.05);the degree of intestinal mucosal damage was reduced,the goblet cells were basically arranged neatly,the inflammatory cell infiltration was reduced;synovial cells proliferation and thickening decreased,with a small amount of inflammatory cell infiltration;the number and evenness of gut microbiota species increased,while the relative abundance of Proteobacteria,Firmicutes,and Bacteroidetes increased(P<0.05,P<0.01),the relative abundance of Desulfobacteria decreased(P<0.01),while the relative abundance of Campylobacter,Lawsonella and Pseudomonas decreased(P<0.01),the relative abundance of Heshanomonas and Herbaspirillum increased(P<0.05,P<0.01);the contents of serum TNF-α,IL-1β and IL-6 significantly decreased(P<0.01).Conclusion Electroacupuncture can significantly improve the symptoms of RA rabbits and reduce the inflammatory reaction in synovial tissue of joint,and the mechanism may be related to the regulation of intestinal flora structure.

【Objective】 To investigate the predictive value of regional cerebral oxygen saturation (rScO2) monitoring during total aortic arch replacement and stent trunk surgery for perioperative neurocognitive disorders (PND) and changes in plasma S100β protein and neuron-specific enolase (NSE) concentrations and their relationship with PND. 【Methods】 Sixty-five Stanford type A aortic dissection patients who planned to undergo total aortic arch replacement and trunk stenting were selected. Their rScO2 values were monitored throughout the operation and recorded after induction (T1), the beginning of CPB (T2), during deep hypothermic circulatory arrest (T3), rewarming to 36℃(T4), CPB stop for 1 hour (T5), and post-operation (T6). After induction (Ta), rewarming to 36℃ (Tb),1 h (Tc), 6 h (Td) and 24 h (Te) after cessation of cardiopulmonary bypass, central venous blood was collected from patients, and the concentrations of S100β protein and NSE in plasma were detected by ELISA. The patients were divided into PND group and non-PND group by the evaluation of MMSE scale at time of before operation, on the day of extubation, and 7 days after operation. 【Results】 The incidence of PND was 44.6%. The rScO2 value at T2 was significantly lower than that at T1 (P<0.05). The rScO2 value of PND group at T3 and T6 was significantly lower than that at T1 and non-PND group (P<0.05). The mean value of rScO2 and the minimum value of rScO2 in PND group were significantly lower than those in non-PND group, while rScO2 %max in PND group was significantly higher than that in non-PND group (P<0.05). The intraoperative critical value of rScO2 %max was >9.89%, the area under curve (AUC) was 0.658 (95% CI: 0.525-0.791, P<0.05), and sensitivity and specificity were 48.3% and 75.0%, respectively. The concentrations of S100β protein and NSE protein in PND group were significantly higher than those in non-PND group at Tc and Td (P<0.01). Compared with Ta, the concentration of S100β protein in PND group was significantly increased at Tc and Td (P<0.001), and the concentration of NSE protein was significantly increased at Tb-Te (P<0.01). CPB time was an independent risk factor for PND. 【Conclusion】 The occurrence of PND after total arch replacement and stenting may be related to the decrease of rScO2 and the increase of S100β protein and NSE protein. Intraoperative rScO2 %max >9.89% can be a potential predictor of PND.

【Objective】 To investigate the role of LIF/LIFR/STAT3 pathway in endometrial receptivity in rats with polycystic ovary syndrome (PCOS). 【Methods】 Forty 21-day-old SD female rats were divided into normal (control) group, model group, sham-operation group, and LIF group with 10 rats in each. The rat model of PCOS was constructed by subcutaneous injection of prasterone sodium sulfate at the back of the neck. The serum levels of testosterone (T), glucose and insulin in each group were detected. The morphological changes of the uterus in each group were observed by HE staining, and the morphological changes of endometrium were measured. Immunohistochemistry, Western blotting, and Real-time fluorescence quantitative PCR(qRT-PCR) were used to determine the protein expression and mRNA expression of LIF and STAT3 in rat endometrium. 【Results】 Compared with control group, the levels of integrin avb3, serum T, insulin and glucose in PCOS rats were significantly increased (P=0.000, P=0.000, P=0.001). Supplementation of exogenous LIF could significantly reduce the levels of integrin avb3, serum T, glucose and insulin in PCOS rats (P=0.000, P=0.002, P=0.003, P=0.007). HE results showed that exogenous LIF could reduce uterine cavity and glandular morphology in PCOS rats and increase the equivalent diameter (P=0.000, P=0.000) and area (P=0.000, P=0.000) of uterine glands and glandular cavity, the ratio of glandular interstitial area (P=0.000), and the average endometrial thickness (P=0.006), with statistically significant differences. Immunohistochemistry, Western blotting, and qRT-PCR results showed that the expression levels of LIF and p-STAT3 protein and mRNA in model group were significantly decreased compared with control group. Compared with model group, the protein and mRNA expressions of LIF and p-STAT3 in LIF group were significantly increased (P<0.05). 【Conclusion】 Exogenous LIF supplementation can improve endometrial receptivity in PCOS rats, and its mechanism is related to the LIF/LIFR/STAT3 pathway.

ObjectiveTo investigate whether Jiedu Huoxue prescription can induce macrophage autophagy and inhibit inflammatory response to stabilize vulnerable plaques of atherosclerosis （AS） by regulating phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodThirty ApoE^-/- mice fed with high-fat diet were randomly assigned into model， low-， medium-， and high-dose （5.35， 10.7， and 21.4 g·kg^-1·d^-1， respectively） Jiedu Huoxue prescription （Chinese medicine）， and rapamycin （2 mg·kg^-1·d^-1） groups. Six ApoE^-/- mice fed with common diet were used as the control group， and 6 C57BL/6J mice fed with common diet as the blank group. The drugs or equal volume of normal saline were administrated by gavage after 7 weeks of modeling， and the treatment lasted for 4 weeks. The serum levels of lipids and inflammatory cytokines ［monocyte chemoattractant protein-1 （MCP-1） and interleukin-6 （IL-6）］ were measured. Hematoxylin-eosin （HE） staining was employed to observe the pathological changes of the vascular wall of the aortic root. Immunohistochemistry was employed to detect the expression of macrophages/monocytes monoclonal antibody （MOMA-2） and α-smooth muscle actin （α-SMA）. Transmission electron microscopy was employed to count the autophagosomes in the aorta， and Western blot to determine the protein levels of Beclin-1， LC3， PI3K， Akt， and mTOR. ResultCompared with the control group， the model group showed elevated serum levels of lipids， MCP-1， and IL-6 （P<0.05）， inhibited expression of MOMA-2 and α-SMA （P<0.05， P<0.01）， up-regulated protein level of Beclin-1 （P<0.05）， and down-regulated protein levels of PI3K， Akt， and mTOR （P<0.05， P<0.01）. The model group presented obvious atherosclerotic plaques on the inner wall of the aorta， infiltration of inflammatory cells in the plaque， thickened and disarranged vascular intima where the plaque was attached， decreased autophagosomes and mitochondria， and destroyed mitochondrial structure. Chinese medicine and rapamycin groups showed lower levels of total cholesterol， triglycerides， low-density lipoprotein cholesterol， MCP-1， and IL-6 （P<0.05）， higher level of high-density lipoprotein cholesterol （P<0.05）， inhibited expression of MOMA-2 and α-SMA （P<0.05， P<0.01）， higher protein levels of Beclin-1 and LC3Ⅱ （P<0.05， P<0.01）， and lower protein levels of PI3K， Akt， and mTOR （P<0.05， P<0.01） than the model group. Moreover， Chinese medicine and rapamycin groups showed only a small number of atherosclerotic plaques on the inner wall of the aorta， reduced infiltration of inflammatory cells and thickness of the blood vessel wall， and increased autophagosomes and autophagic lysosomes. ConclusionJiedu Huoxue prescription can improve lipid metabolism， enhance macrophage autophagy， and reduce AS-induced inflammation to improve the stability of vulnerable plaques in AS mice by inhibiting the PI3K/Akt/mTOR signaling pathway.

Objective To explore the role and mechanism of microRNA-204(miR-204) carried by the exosomes of human umbilical cord-derived mesenchymal stem cells(hUC-MSC) in regulating the polarization of macrophages in a mouse model of myocardial ischemia-reperfusion(I/R) injury. Methods After the hUC-MSCs were isolated,cultured,and identified,their adipogenic and osteogenic differentiation capabilities were determined.The exosomes of hUC-MSCs were separated by ultracentrifugation,and the expression of CD81,CD63,tumor susceptibility gene 101(Tsg101),and calnexin in the exosomes was determined by Nanoparticle Tracking Analysis software,transmission electron microscopy,and Western blotting.Three groups(hUC-MSC,miR-204 mimic,and negative control) were designed for the determination of the expression of miR-204 in the cells and their exosomes by qRT-PCR.The C57BL/6J mice were randomly assigned into a sham operation group,an I/R group,a hUC-MSC exosomes group,a negative control group,and a miR-204 mimic group.Except the sham operation group,the I/R model was established by ligating the left anterior descending artery.The echocardiography system was employed to detect the heart function of mice.HE staining was employed to observe the pathological changes of mouse myocardium.ELISA was employed to determine the levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),arginase 1(Arg-1),and IL-10 in the myocardial tissue.After the macrophages of mouse myocardial tissue were isolated,flow cytometry was employed to determine the expression of CD11c and CD206,and ELISA to measure the levels of IL-1β,TNF-α,Arg-1,and IL-10 in the macrophages. Results hUC-MSCs had adipogenic and osteogenic differentiation capabilities,and the exosomes were successfully identified.Compared with the negative control group,the miR-204 mimic group showed up-regulated expression of miR-204 in hUC-MSCs and their exosomes(P<0.001,P<0.001).Compared with the sham operation group,the modeling of I/R increased the left ventricular end-diastolic diameter(LVEDD)(P<0.001),left ventricular end-systolic diameter(LVESD)(P<0.001),myocardial injury score(P<0.001),and the levels of IL-1β(P<0.001),TNF-α(P<0.001),and CD11c(P<0.001).Meanwhile,it lowered the left ventricular ejection fraction(LVEF)(P<0.001),left ventricular fractional shortening(LVFS)(P<0.001),Arg-1(P<0.001),IL-10(P<0.001),and CD206(P<0.001).Compared with those in the I/R group,the LVEDD(P<0.001),LVESD(P<0.001),myocardial injury score(P<0.001),and the levels of IL-1β(P<0.001),TNF-α(P=0.010),and CD11c(P<0.001) reduced,while LVEF(P<0.001),LVFS(P<0.001),and the levels of Arg-1(P<0.001),IL-10(P=0.028),and CD206(P=0.022) increased in the hUC-MSC exosomes group.Compared with those in the negative control group,the LVEDD(P<0.001),LVESD(P<0.001),myocardial injury score(P=0.001),and the levels of IL-1β(P=0.048),TNF-α(P<0.001),and CD11c(P=0.007) reduced,while the LVEF(P<0.001),LVFS(P<0.001),and the levels of Arg-1(P<0.001),IL-10(P=0.001),and CD206(P=0.001) increased in the miR-204 mimic group. Conclusion The hUC-MSC exosomes overexpressing miR-204 can inhibit the polarization of macrophages in the I/R mouse model to M1-type and promote the polarization to M2-type.
Animals ; Humans ; Mice ; Disease Models, Animal ; Exosomes/pathology* ; Interleukin-10/metabolism* ; Macrophages ; Mesenchymal Stem Cells ; Mice, Inbred C57BL ; MicroRNAs/genetics* ; Myocardial Reperfusion Injury ; Osteogenesis ; Stroke Volume ; Tumor Necrosis Factor-alpha/metabolism* ; Umbilical Cord/pathology* ; Ventricular Function, Left

Objective:To explore the correlation between serum soluble T cell immunoglobulin-mucin molecule-3 (sTim-3), galactose lectin-9 (Galectin-9) and inflammatory factors interferon-γ (IFN-γ), interleukin-4 (IL-4) in patients with brucellosis.Methods:Ninety patients with brucellosis admitted to Yantai Yeda Hospital from January 2016 to December 2018 were selected as the observation group, of whom 49 cases in acute stage and 41 cases in chronic stage. Another 50 healthy subjects who underwent health examination during the same period were selected as the control group. Five milliliter of fasting venous blood samples were collected from patients with brucellosis and healthy subjects, and enzyme-linked immunosorbent assay (ELISA) was used to detect sTim-3 and Galectin-9 levels, and cytometric bead array (CBA) to detect IFN-γ and IL-4 levels; Pearson correlation test was used to correlate serum levels of sTim-3, Galectin-9, sTim-3/Galectin-9 and inflammatory factors IFN-γ, IL-4 in patients with brucellosis.Results:The levels of sTim-3 [(18.28 ± 5.38), (7.01 ± 1.97) ng/L], Galectin-9 [(2.79 ± 0.94), (1.67 ± 0.23) ng/L], sTim-3/Galectin-9 (6.55 ± 2.96, 4.20 ± 0.67), IFN-γ [(5.99 ± 1.94), (10.09 ± 2.03) ng/L], IL-4 [(7.34 ± 3.13), (3.71 ± 1.76) ng/L] were significantly different between the observation group and control group ( t = 14.271, 8.276, 5.527, 11.785, 7.556, P < 0.05). In the observation group, the levels of sTim-3 [(22.09 ± 3.13), (14.21 ± 3.42) ng/L], Galectin-9 [(2.98 ± 0.67), (2.41 ± 0.74) ng/L], sTim-3/Galectin-9 (7.41 ± 1.07, 5.90 ± 1.72), IFN-γ [(3.78 ± 1.65), (7.02 ± 1.73) ng/L], and IL-4 [(9.17 ± 2.04), (5.67 ± 1.54) ng/L] were statistically significant difference between patients with acute and chronic brucellosis ( t = 11.402, 3.833, 5.084, 9.075, 9.037, P < 0.05). Pearson correlation test showed that the levels of sTim-3, Galectin-9, and sTim-3/Galectin-9 in patients with brucellosis were negatively correlated with IFN-γ level ( r = - 0.568, - 0.531, - 0.519, P < 0.05), and positively correlated with IL-4 level ( r = 0.517, 0.524, 0.536, P < 0.05). Conclusion:The expressions of sTim-3 and Galectin-9 in serum of brucellosis patients are high, and they are significantly correlated with IFN-γ and IL-4 levels, suggesting that sTim-3 and Galectin-9 maybe related to the occurrence and development of brucellosis.

Objective To analyze clinical outcome of high grade squamous intraepithelial lesion (HSIL) within 24 months after loop electrosurgical excision procedure (LEEP),and to explore risk factors of recurrent cervical HSIL,the risk of progress into cervical cancer and methods of follow-up.Methods This retrospective study was carried out on 1 005 patients who underwent LEEP,diagnosing with HSIL after LEEP from January 2011 to December 2013 at Obstetrics and Gynecology Hospital Affiliated to Fudan University to confer the difference between non-recurrent group and recurrent group 24 months after the LEEP conization.Patients were followed with ThinPrep cytologic test (TCT),high risk HPV (HR-HPV) test,colposcopy guided biopsy.Results A total of 1 005 cases were enrolled in this study with HSIL in the LEEP specimen,no residual HSIL in the 6-month follow up,and have follow up records in 24 months after LEEP.HSIL recurred in 5 cases,microinvasive carcinoma in 1 case,low grade squamous intraepithelial lesion (LSIL) in 17 cases at 12 months follow-up.HSIL recurred in 8 cases,LSIL in 11 cases,adenocarcinoma in situ in 1 case,and invasive cervical carcinoma in 1 case in Ⅰ b1 stage at 24 months after LEEP.The recurrence rate was 1.3％ (13/1 005),and the progression rate was 0.3％ (3/1 005).There was no significant difference in age,length,circumference and width of LEEP between recurrent and non-recurrent patients (P ＞ 0.05).The recurrence rate was highest in the endocervical positive margin group with 3/16,which was higher than ectocervical positive margin and negative margin (P＜0.01,P=0.040,respectively).The recurrence rate of endocervical positive margin group and fibrous interstitial positive margin group showed no significant difference (P=0.320).There was no significant difference between ectocervical positive margin and negative margin [2.8％ (2/72) vs 0.7％ (6/882),P=0.117].Postoperative cytological examination combined with HR-HPV detection has a high sensitive and high negative predictor value of HSIL recurrence with both 100.0％.Multivariate logistic regression analysis showed that positive endocervical positive margin,abnormal follow-up cytological examination and positive HR-HPV after LEEP were independent factors affecting recurrence of HSIL patients after LEEP (P＜0.05).Conclusions Age,length,circumference and width of LEEP have no effect on recurrence within 24 months after HSIL.The high risk factors for HSIL recurrence within 24 months after LEEP in HSIL patients include:positive HPV,abnormal cytology,and positive endocervical positive margin.Colposcopy biopsy and endocervical curettage have important role in diagnosing HSIL recurrence and progression.

Objective To observe the effect of early bundle therapy on prognosis of patients with sepsis/septic shock and analyze the risk factors for death.Methods A retrospective cohort study was conducted to select patients with sepsis/septic shock at the Second Soochow University Hospital betweenJanuary 1,2016,and December 31,2016.Data pertaining to demographic variables,compliance rate of bundle therapy,and incidence of organ failure were collected.Patients were categorized into the nonsurvivor or survivor groups based on 28-day mortality.Logistic regression analysis was used to identify risk factors for 28-day mortality.Results Totally 118 sepsis/septic shock patients were included in the analysis;28-day mortality was 32.2％.Compared to the survivor group,patients in the non-survivor group were more likely to have chronic heart dysfunction and cerebrovascular disease,lower lactate clearance,lower 6-h compliance rate of bundle therapy and higher incidence of failure of one or ＞2 organs.Age,leukocyte,blood urea nitrogen,creatinine,brain natriuretic peptide,sequential organ failure score and acute physiological and chronic health scores Ⅱ on admission,and lactate after bundle therapy were higher than that of the survivor group.Logistical regression analysis showed that age ≥ 75 years [odds ratio (OR)1.012],6-h lactate clearance ＜30％ (OR=1.122),chronic heart failure (OR=1.741),failure of ＞2 organs (OR=1.769),and 6-h compliance rate of bundle therapy (OR=1.958) were independent risk factors for 28-day mortality.Conclusions Patients with sepsis/septic shock need early diagnosis and resuscitation to improve the compliance rate of bundle therapy and reduce the mortality.