Organ preservation fluid could mitigate cold ischemia injury and maintain normal function of the grafts. At present, how to reduce a series of injury caused by cold ischemia of donor liver and improve the preservation quality of grafts are the hot and challenging spots in this field. Currently, preservation fluid in clinical practice has not achieved ideal preservation effect, especially for the protection of marginal donor organs. In the context of severe donor shortage, the key solution is still to explore the optimal preservation protocol for donor liver to prevent grafts from cold ischemia injury. In this article, the mechanism of donor liver injury during cold ischemia, the classification and evolution of donor liver preservation fluid were summarized, the development direction and challenges of donor liver preservation fluid were discussed, aiming to provide novel ideas and references for the research and development of donor liver preservation fluid.

Objective To investigate the safety and efficacy of different surgical sequences in the ultrasound-guided endovenous microwave ablation combined with foam sclerotherapy for the treatment of primary great saphenous varicose veins.Method A total of 80 patients with great saphenous varicose vein admitted in the affiliated hospital of Jiangsu University,from January 2022 to January 2023 were selected.Patients were divided into observation group and control group according to different operation order,with 40 cases in each group.The control group was treated with ultrasound-guided micro wave ablation of the main saphenous vein was performed first,followed by superficial calf vein foam sclerotherapy injection and local small incision point extraction,and the observation group was treated with superficial calf vein foam sclerotherapy injection and local small incision point extraction first,followed by ultrasound-guided microwave ablation of the main saphenous vein was performed.Perioperative relevant indicators at the 1st week of the two groups were counted,and the incidence of hematoma,ecchymosis,induration,skin burn,thrombotic superficial phlebitis,and endovenous heat induced thrombosis at the 1st week after surgery.The venous clinical severity score and chronic venous insufficiency quality of life at the 3rd and 6th month after surgery were compared between the two groups.VCSS and CIVIQ were used to evaluate the postoperative recovery of patients with varicose veins.Six months after the operation,the recurrence rate of great saphenous vein was compared by color Doppler ultrasonography.Result The operation time of the two groups was(68.13±3.34)min and(66.83±3.19)min,respectively.The intraoperative blood loss was(15.35±2.63)ml and(14.83±2.66)ml,respectively.The underground activity time was(14.35±3.34)hours and(13.60±2.63)hours,respectively.The length of hospitalization was(2.93±0.52)days and(3.15±0.61)days,respectively.There was statistical significance between the two groups(P＜0.05).The preoperative VCSS of the two groups were 4.08±1.37 and 4.23±1.33,respectively,3 months after surgery were 3.00±0.59 and 3.03±0.61,respectively,and 6 months after surgery were 2.20±1.17 and 2.35±0.96,respectively.The preoperative CIVIQ of the two groups were 79.63±5.41 and 80.03±7.44,respectively,3 months after operation was 69.90±2.98 and 70.43±3.55,respectively,the 6-month CIVIQ was 59.05±3.79 and 58.00±4.66,respectively.There was no statistical significance between the two groups(P＞0.05).The incidence of adverse events[hematoma(0 vs 0),ecchymosis(12.5％vs 15.0％),sclerosis(10.0％vs 7.5％),skin burns(0 vs 0),thrombosed superficial phlebitis(12.5％vs 17.5％),and thermal ablation-induced thrombosis(10.0％vs 5.0％)]in the patients of the two groups in the 1-week period after the procedure were compared,and the difference were statistically non-significant(P＞0.05).Comparison of trunk recanalisation rate(5.0％vs 2.5％)at 6 months after surgery,the difference was not statistically significant(P＞0.05).Conclusion There is no significant difference in the efficacy of the two procedures in the treatment of primary saphenous varicose veins,with a high degree of safety,both of which are worthy of clinical promotion.

New drug discovery is under growing pressure to satisfy the demand from a wide range of domains, especially from the pharmaceutical industry and healthcare services. Assessment of drug efficacy and safety prior to human clinical trials is a crucial part of drug development, which deserves greater emphasis to reduce the cost and time in drug discovery. Recent advances in microfabrication and tissue engineering have given rise to organ-on-a-chip, an in vitro model capable of recapitulating human organ functions in vivo and providing insight into disease pathophysiology, which offers a potential alternative to animal models for more efficient pre-clinical screening of drug candidates. In this review, we first give a snapshot of general considerations for organ-on-a-chip device design. Then, we comprehensively review the recent advances in organ-on-a-chip for drug screening. Finally, we summarize some key challenges of the progress in this field and discuss future prospects of organ-on-a-chip development. Overall, this review highlights the new avenue that organ-on-a-chip opens for drug development, therapeutic innovation, and precision medicine.

Objective To optimize the culture and fermentation conditions of the Penicillium aurantiocandidum Z12 strain, a fungal strain with molluscicidal actions against Oncomelania hupensis, so as to provide the basis for the research and development of molluscicidal active substances from the P. aurantiocandidum Z12 strain and its fermentation broth and large-scale fermentation. Methods The carbon source, nitrogen source and mineral salts were identified in the optimal culture medium for the P. aurantiocandidum Z12 strain with a single-factor experiment to determine the best fermentation condition for the P. aurantiocandidum Z12 strain. Factors that significantly affected the growth of the P. aurantiocandidum Z12 strain were identified using the Plackett-Burman design, and the best range of each factor was determined using the steepest climb test. Response surface analyses of temperature, pH value, seeding amount and liquid-filling quantity were performed using the Box-Behnken design to create a regression model for fermentation of the P. aurantiocandidum Z12 strain to identify the optimal culture medium. Results Single-factor experiment preliminarily identified the best culture medium and conditions for the P. aurantiocandidum Z12 strain as follows: sucrose as the carbon source at approximately 20 g/L, tryptone as the nitrogen source at approximately 5 g/L, K₂HPO₄ as the mineral salt at approximately 5 g/L, initial pH at approximately 8, temperature at approximately 28 °C, seeding amount at approximately 6%, and liquid-filling quantity at approximately 50 mL/100 mL. Plackett-Burman design showed that factors that significantly affected the growth of the P. aurantiocandidum Z12 strain included temperature (t = −5.28, P < 0.05), seeding amount (t = 5.22, P < 0.05), pH (t = −4.30, P < 0.05) and liquid-filling quantity (t = −4.39, P < 0.05). Steepest climb test showed the highest mycelial growth at pH of 7.5, seeding amount of 8%, and liquid-filling quantity of 40 mL/100 mL, and this condition was selected as the central point of response surface analysis for the subsequent optimization of fermentation conditions. Response surface analyses using the Box-Behnken design showed that the optimal conditions for fermentation of the P. aurantiocandidum Z12 strain included sucrose at 15 g/L, tryptone at 5 g/L, K₂HPO₄ at 5 g/L, temperature at 28.2 °C, pH at 7.5, seeding amount at 10%, and liquid-filling quantity at 35.8 mL/100.0 mL, resulting in 0.132 g yield of the P. aurantiocandidum Z12 strain. Conclusion The optimal culture condition for the P. aurantiocandidum Z12 strain has been identified, and the optimized culture medium and fermentation condition may effectively improve the fermentation yield of the P. aurantiocandidum Z12 strain.

Tryptophan 2,3-dioxygnease 2 (TDO2) is specific for metabolizing tryptophan to kynurenine (KYN), which plays a critical role in mediating immune escape of cancer. Although accumulating evidence demonstrates that TDO2 overexpression is implicated in the development and progression of multiple cancers, its tumor-promoting role in esophageal squamous cell carcinoma (ESCC) remains unclear. Here, we observed that TDO2 was overexpressed in ESCC tissues and correlated significantly with lymph node metastasis, advanced clinical stage, and unfavorable prognosis. Functional experiments showed that TDO2 promoted tumor cell proliferation, migration, and colony formation, which could be prevented by inhibition of TDO2 and aryl hydrocarbon receptor (AHR). Further experimentation demonstrated that TDO2 could promote the tumor growth of KYSE150 tumor-bearing model, tumor burden of C57BL/6 mice with ESCC induced by 4-NQO, enhance the expression of phosphorylated AKT, with subsequent phosphorylation of GSK3

Objective To investigate the characteristics and feasibility of genipin-crosslinked amniotic membrane(AM) as bio-scaffold.Methods Human umbilical cord mesenchymal stem cells (hUCMSCs) were isolated from fresh umbilical cord and cultured by adherent method.The expressions of PE-CD34,PE-CD45,PE-CD90,FITC-105 and FITC-Oct-4,the markers of hUCMSCs,were detected by flow cytometry.Alizarin red and oil red O staining were performed to identify the cells after adipogenesis and osteogenesis induction on the third-generation cells.Human AMs were treated at 37 ℃ and 45 ℃ by 0.5％ and 1％ genipin solution for 24,36 and 48 hours respectively,and the mechanical properties of AM in each group were measured and compared.The hUCMSCs were divided into only hUCMSCs culture group,fresh AM group,crosslinked AM group,gelatin group and crosslinked AM+gelatin group,and the cells were cultured in the corresponding medium.The content of hydroxyproline among the groups was detected with hydroxyproline kit,and proliferation of the cells (absorbance) was assayed by MTT method to evaluate the biological compatibility of crosslinked AM.Results The maximum tensile displacement of the crosslinked-AM by 0.5％ and 1％ genipin was (8.31±0.43)mm and (4.49±0.37)mm respectively,and those after crosslinked with 0.5％ genipin under the 37 ℃ and 45 ℃ for 24 hours was (9.89±1.09)mm and (5.39±0.59)mm,respectively,showing a significant difference between them (t =6.389,P＜0.05).The maximum tensile displacement of the crosslinked-AM was gradually decreased as the lapse of crosslinking time,and an insignificant difference was found among 24,36 and 48 hours after 0.5％ genipin treatment under the 37 ℃ (P＞0.05).The loading force of the crosslinked-AM was significantly higher in the 1％ genipin treated group than that in the 0.5％ genipin treated group (P＜0.05),and the loading force of the AM was significantly increased in 45 ℃,0.5％ genipin,24 hours crosslinked group compared with the 37 ℃,0.5％ genipin,24 hours crosslinked group (t =5.528,P＜0.05).The content of hydroxyproline in the AM was (1.28±0.36),(2.03 ±0.49) and (2.11 ±0.10) mg/g in the 1％ genipin crosslinked AM group,0.5％ genipin crosslinked AM group and fresh AM group,respectively,and the content of hydroxyproline in the AM in the 1％ genipin group was significantly lower than that in the 0.5％ genipin group in the fresh AM group (both at P＜0.05).The proliferative values of the hUCMSCs were significantly lower in the only hUCMSCs culture group,fresh AM group and gelatin group were significantly reduced in comparison with the crosslinked AM group and crosslinked AM+gelatin group (all at P＜0.05).There was no significant difference in the proliferative values of the hUCMSCs between crosslinked AM group and crosslinked AM+gelatin group (P＞0.05).Conclusions Different crosslinked temprature,crosslinking period and concentration of genipin impact the mechanical properties of AM.Crosslinked AM with genipin is feasible as a carrier scaffold of artificial cornea because of less tissue toxicity and better plasticity.

Objective The 18F-ML-10 PET/CT apoptosis imaging was taken in the early Parkinson's disease (PD) rat models.The feasibility of diagnosis of PD with 18F-ML-10 PET/CT apoptosis imaging is explored.Methods Twenty adult healthy male Sprague-Dawley rats were randomly divided into control group and PD model group (n=l 0).Intrastriat administration of 6-hydroxy dopamine (6-OHDA) was performed to induce progressive and retrograde degenerative changes in the substantial nigra of neurons (PD models).One week after apomorphine inducement,the rotational behaviors of the rats were evaluated.~C-CFT PET/CT imaging and 18F-ML-10 PET/CT were performed to observe the dopamine transport protein expression in the corpus striatum and apoptosis of dopaminergic neurons in the substantia nigra.Immunofluorescence staining of anti-tyrosine hydroxylase (TH) antibody was performed to evaluate the survival of dopaminergic cells in the compact part of substantia nigra.TUNEL was performed to evaluate the apoptosis of dopaminergic cells in the compact part of substantia nigra.Nissl staining was performed to detect the cellular morphology.Results One week after PD modeling,the rats in the experimental group obviously rotated to the contralateral,and the average rotation speed was (4.52 ±1.03) r/min.The ratio of 11C-CFT between the right and left striatum of rats in the experimental group was 0.556±0.017,which was significantly lower than that of the control group (0.998±0.013,P＜0.05).The radioactivity ratio of'8F-ML-10 between the right and left substantia nigra of rats was 1.722±0.083,which was significantly higher than that of the control group (1.024±0.056,P＜0.05).Immunofluorescence showed that the ratio of TH-positive neurons between the right and left compact part of substantia nigra in rats of the experimental group was 0.528 ±0.012,which was significantly lower than that of the control group (1.036±0.030,P＜0.05).TUNEL showed that the number of dopaminergic neuronal apoptosis in the experimental group was 43.200±2.507,which was significantly larger than that of the control group (1.400±0.427,P＜0.05).Conclusion PD is associated with apoptosis of dopaminergic neurons;18F-ML-10 PET/CT imaging can be used to diagnose PD in the early-stage.

OBJECTIVETo observe the effect of Shenshuai Yingyang Capsule (SSYYJN) in ameliorating muscle atrophy in rats with chronic renal failure (CRF) and explore the role of Wnt7a-Akt/mTOR signal pathway in mediating this effect.

METHODSMale rats were randomly assigned to 5/6 nephrectomy group and sham-operated group, and the former group was further randomly divided into CRF model group, KA group, and SSYYJN group. The size of anterior tibia muscle was examined microscopically with HE staining. Protein synthesis in the soleus muscle was investigated by (14)C-phenylalanine experiment, and the expression of Wnt7a, frizzled-7, phospho-Akt, phospho-mTOR and GAPDH were detected with Western blotting.

RESULTSThe body weight, the wet and dry weight, cross-sectional area, and muscle protein synthesis of the anterior tibia muscles, and expressions of the proteins in the Wnt7a/Akt signaling pathway all increased significantly in SSYYJN and KA groups as compared with those in the model group.

CONCLUSIONSSYYJN can effectively improve muscle atrophy in the rat model of CRF possibly by reversing the reduction in the expressions of Wnt7a/Akt signaling pathway proteins in the skeletal muscles.

Animals ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; Kidney Failure, Chronic ; complications ; Male ; Muscle Proteins ; biosynthesis ; Muscle, Skeletal ; drug effects ; Muscular Atrophy ; drug therapy ; Nephrectomy ; Proto-Oncogene Proteins ; metabolism ; Rats ; Signal Transduction ; TOR Serine-Threonine Kinases ; metabolism ; Wnt Proteins ; metabolism

Objective To study the effects of endoplasmic reticulum stress on liver damage in young rats fed with high-fat diet.Methods We divided 48 male weaned young rats randomly into high-fat diet group and control group,which were separately fed with high-fat diet and normal diet.After feeding 8,12 and 1 6 weeks,the body weight and visceral fat of the rats were measured.The serum liver function was measured.The morphology of livers was observed by HE and transmission electron microscopy. The mRNA expressions of ATF6 and GRP78 in hepatocytes were measured with RT-PCR.Results ① The body weight and visceral fat weight of rats in high-fat diet group increased compared with those in control group (P <0.05).② Alanine aminotransferase and aspartate aminotransferase in high-fat diet group increased slightly over time (P >0.05);alanine aminotransferase at week 1 6 was increased significantly compared with that in controls (P < 0.05 ).③ Liver cells in high-fat diet group had steatosis at week 8 and the steatosis became more serious between week 12 and week 1 6.④ In high-fat diet group at week 8 there were a large number of lipid droplets in the cytoplasm,and the cell structure was close to that of normal cells;rough endoplasmic reticulum was nearly normal and the ribosome was visible.At week 12 and week 1 6,besides a large number of lipid droplets,we could also see that some substances with line-like structure deposited in rough endoplasmic reticulum pool.⑤ The expressions of ATF6 and GRP78 mRNA in hepatocytes in high-fat group at weeks 8, 12 and 1 6 were significantly increased compared with those in control group (P <0.05). Conclusion High-fat diet in infants can cause visceral fat accumulation,fatty degeneration of hepatocytes and liver injury.ATF6-mediated endoplasmic reticulum may be closely related to the liver injury which results from highfat diet.

Objective To establish an in vitro model of cultured mouse testis using rotary aerobic culture. Methods Rotary aerobic incubation with optimized culture conditions was used for in vitro culture of mouse testis, and the morphology of the cultured testicular tissues was compared with that cultured in Transwell chambers. The changes in the testicular tissue structure were examined using HE staining, and the cell proliferation was assessed with BrdU staining. Testosterone concentrations in the culture medium were tested with radioimmunoassay and the expression of the functionally related proteins in the testis was detected using immunohistochemistry. Results The testicular tissue cultured by optimized rotary aerobic culture presented with more intact histological structure with the size of the testis ranged from 0.3 to 0.8 mm3. In the two culture systems, the prolifeation index of the spermatogonia increased and that of Sertoli cells decreased with time, and such changes in spermatogonia and Sertoli cell proliferation indices became statistically significant at 3 days (P<0.05) and 5 days (P<0.05) of culture, respectively, as compared with those at 1 day. The concentration of testoerone in the culture media decreased significantly with incubation time (P<0.05). At 3 days of culture, the protein expression of 3β-hydroxysteroid dehydrogenase, cytochrome P450 17α-hydroxylase and cholesterol side-chain cleavage enzyme was detected in Leydig cell cytoplasm and vimentin expression in Sertoli cell cytoplasm. Conclusion An in vitro model of cultured mouse testis has been successfully established using rotary aerobic incubation.