BACKGROUND:Cartilage scaffold plays an important role in articular cartilage injury, but it is stil restricted in practice, leading to the repair failure using tissue-engineered cartilage. OBJECTIVE:To explore the preparation method of cartilage scaffold with directional structure and its repairing effect in cartilage injury of rabbits. METHODS:Directional cartilage scaffolds were prepared by temperature gradient induced phase separation technique. The cross-linking of cartilage scaffold was completed with genipin. The physical and mechanical properties of scaffolds were determined. Forty-eight white rabbits were modeled into exercised-induced articular cartilage injury and equivalently randomized into two groups, fol owed by implantation of non-directional or directional scaffolds. Samples were col ected at postoperative 6, 12 and 24 weeks to compare the repair effect of two scaffolds on rabbit joint injury. RESULTS AND CONCLUSION:Scanning electron microscope showed that the regular microstructures on the longitudinal section of directional scaffolds were aligned in the same direction, while the longitudinal and transverse planes of the non-directional scaffolds showed porous honeycomb structures. The elastic modulus of the directional scaffold was significantly higher than that of the non-directional scaffold (P<0.05). O’Driscol histological scores in the directional scaffold group were significantly higher than those in the non-directional scaffold group at 6, 12 and 24 weeks postoperatively (P<0.05). At postoperative 24 weeks, in the directional scaffold group, the three-dimensional reconstruction of the rabbit defect region was flat and smooth, and coronal two-dimensional scanning showed the cartilage thickness of the newborn cartilage was very close to that of the normal bone;the cartilage surface in the non-directional rabbits was flat under three-dimensional CT, and coronal two-dimensional scanning showed that the repaired cartilage was almost the same as the normal cartilage. Hematoxylin-eosin staining showed that at 24 weeks postoperatively, the defect site disappeared and the cartilage cel s arranged in the vertical direction in the directional scaffold group. There were typical cartilage lacunae at the defect site in the non-directional scaffold group at 24 weeks postoperatively. Our findings suggest that the directional cartilage scaffold prepared using temperature gradient induced phase separation technique holds good physical and mechanical properties, and its implantation into the rabbit articular cartilage injury induced by exercise can promote the injury repair to obtain ideal mechanical properties.

Longnon-codingRNAs(lncRNAs)areagroupofRNAslongerthan200bpwithoutprotein-coding capacity and play an important role in epigenetics , transcription regulation and post-transcription regulation .Recently, studies have shown that dysregulation of lncRNAs caused cell cycle disorders , and abnormal activities of cell apoptosis and autophagy , contributing to a variety of diseases .Therefore , the role of lncRNAs in regulating cell death is expected to lead to the discovery of potential novel diagnostic markers and therapeutic targets .

The effect of cognition need on information seeking behaviors of information users and the difference in behaviors of information users were investigated in 184 undergraduates from the same grade with different back-grounds of subjects using the cognition need scale and undergraduates information-seeking behaviors scale ,respec-tively.The data were analyzed by descriptive statistical analysis, correlation analysis, rank test, and multivariate linear regression analysis , respectively , showing that the cognition need is significantly correlated with the informa-tion seeking behaviors of undergraduates in terms of information evaluation, information searching, information management and information use .

Objective To prepare mouse model with heat stress and determine its pathological changes of the lung and brain during heat stress. Methods BALB/c mouse were randomly (random number) divided into two groups, control group and heat stress group. The animals in the control group were sham- heated at a temperature of ( 25 ± 0.5) ℃ and humidity of (35 ± 5 ) %. The animals of heat stress group were placed in a prewarmed incubator maintained at (35.5 ± 0.5) ℃ and relative humidity of (60 ± 5) %. Rectal temperature (Tc) was monitored, and when Tc respectively reached 39 ℃, 40 ℃ , 41 ℃ and 42 ℃, those study animals were killed. The other animals were removed from the incubator and allowed to cool at an ambient temperature of (25 ±0. 5)℃ and humidity of (35 ±5)% , respectirvely for 12 and 24 hrs when Tc reached 41 ℃ , and for 6 hrs when Tc reached 42 ℃. The lung and brain of all the animals were isolated. Hematoxylin and eosin stain and light microscope were used to detect their pathological changes. Results All the animals displayed uniform response to the heat stress. Low degree of heat stress could induced obviously pathological changes of the lung, progressively greater damage to lung with further congestion of lung matrix, asystematic hemorrhage of alveolar space, abscission of alveolar epithelial cell and disappear of pulmonary alveolus tissue structure were detected with the rise of Tc to 42 ℃. However, absorption of congestion and hemorrhage and recovery of pulmonary alveolus tissue structure could also be seen with cooling at ambient temperature. With low degree of heat stress, the brain only showed moderate edema. Neuronal denaturation and necrosis were detected when Tc reached to 42 ℃. Interestingly, the lesions of brain further aggravated even through cooling treatment after Tc reached to 42 ℃ , but recovery could been observed after cooling treatment followed with Tc of 41 ℃. Conclusions The pathological changes of the lung and brain showed distinctive lesions to heat stress and cooling treatment, and these changes were correlated with the timing and time of cooling treatment, which provide the experimental basis to further study the mechanisms between the heatstroke and multiple organ dysfunction syndrome (MODS).

Objective To investigate the effect of mitogen-activated protein kinases (MAPKs) activation on the heat stressinduced apoptosis of pulmonary microvascular endothelial cells (PMVECs).Methods A mouse model of severe heat stroke was made and TUNEL and immunohistochemistry were employed to detect lung tissue damage.MACS separation was used for isolation of neonatal PMVECs,and TUNEL was utilized to detect the apoptosis of PMVECs.Western blotting was used for determining the MAPKs activation during heat stress recovery (0,2,6h).The monolayer permeability of endothelial cells was detected in terms of transmembrane resistance (TEER) and horseradish peroxidase (HRP).Cells were pretreated with MAPKs activation inhibitors to examine the effect of heat stress on the monolayer cell permeability and apoptosis.Results In mice with severe heat stroke,extensive apoptosis of PMVECs was found in their pulmonary tissues.TUNEL revealed that the number of apoptotic cells increased over time during heat stress recovery period and heat stress could activate MAPKs in PMVECs.Compared with heat stress group,in the cells pretreated with p38 or ERK activation inhibitor PD98059 and SB203580,the monolayer permeability and apoptosis increased while in cells pretreated withJNK inhibitor SP600125,the cellular permeability and apoptosis decreased.Conclusion In mice with severe heat stoke,PMVECs might experience apoptosis and p38 and ERK could inhibit apoptosis while JNK could promote apoptosis.

OBJECTIVETo investigate the protective effect of ulinastatin (UTI) against acute lung injury induced by heatstroke in mice.

METHODSSixty C57/BL6 mice were randomly divided into 6 groups, with 10 mice in each: control group, heatstroke group, UTI pretreatment group, saline pretreatment group, UTI post-treatment group, saline post-treatment group. The control mice were housed at a controlled room temperature of (22∓1) degrees; celsius, and the other groups were placed inside a temperature and humidity controlled chamber pre-set at 37 degrees; celsius and 60%. The two UTI groups were intraperitoneally injected with UTI at 5×10(4) U/kg 10 min before or after heat stress, and the two saline groups were given then equal amounts of saline in the same manner. The core body temperature of mice was monitored by a mercury thermometer every 30 min in the first 1.5 h during heating. The core temperature was measured, then every 15 min until it reached 42.7 degrees; celsius, which was taken as the onset of heatstroke. The animals were allowed to recover passively at ambient temperature for 6 h. The lung histopathological changes, protein concentration in BALF, lung wet/dry weight ratios, lung water content, and pulmonary microvascular permeability were assayed after 6 h of recovery at 37 degrees;celsius.

RESULTSCompared with the control group, the heatstroke model group and two saline groups displayed more severe lung damage and pathological morphology changes, and the lung wet/dry weight ratio, protein concentration in BALF, lung water content and pulmonary microvascular permeability were also significantly increased. These effects were significantly alleviated in UTI treated group. Pretreat ment with UTI significantly prolonged the time to Tc≥42.7 degrees; celsius but had no effect on lung injury induced by heatstroke.

CONCLUSIONUTI can reduce the pulmonary edema and inflammatory exudation in acute lung injury caused by heatstroke.

Acute Lung Injury ; drug therapy ; physiopathology ; Animals ; Body Temperature ; Bronchoalveolar Lavage Fluid ; chemistry ; Edema ; prevention & control ; Glycoproteins ; therapeutic use ; Heat Stroke ; physiopathology ; Lung ; pathology ; Mice ; Mice, Inbred C57BL

OBJECTIVE: To detect genetic mutations in a child with late-onset hemophagocytic lymphohistiocytosis (HLH). METHODS: Clinical data of an 8-year-5-month-old girl with recurrent HLH and severe central nervous system disease was analyzed. Next-generation sequencing was used to detect mutation in the exons and adjacent introns of 17 genes associated with HLH. Suspected mutations were confirmed by Sanger sequencing. Influence of mutations on protein function was predicted with SIFT and PolyPhen-2 software. RESULTS: The child was found to carry compound heterozygous mutations of the PRF1 gene. Among these, the c.1349C>T (p.Thr450Met) mutation, with a SIFT predictive value of -4.921 (Deleterious variant) and a PolyPhen-2 predictive value of 1.000 (Probably damaging), was inherited from her father and known to be pathogenic. The c.1273dupT (p.Trp425fsX457) mutation was inherited from her mother and previously unreported, which resulted in the deletion of almost the entire C2 domain (amino acid residues 413 to 540) and carboxyl terminal of perforin, which seriously affected the function of the protein. CONCLUSION The c.1349C>T (p.Thr450Met) and c.1273 dupT (p.Trp425fsX457) compound heterozygous mutations of the PRF1 gene probably underlie the disease in this patient.
Central Nervous System Diseases ; Child ; Exons ; Female ; Humans ; Lymphohistiocytosis, Hemophagocytic ; Mutation ; Perforin

OBJECTIVETo investigate the molecular mechanisms of continuous venovenous hemofiltration (CVVH) combined with ulinastatin (ULI) (CVVH-ULI) for the treatment of septic shock.

METHODSHuman umbilical endothelial cells (HUVECs) were incubated with serums isolated from normal healthy people (control), septic shock patients treated with conventional therapy (CT) or treated with CVVH combined with ULI (CVVH-ULI). Endothelial permeability was evaluated by the leakage of FITC-labeled albumin. The morphological changes of F-actin was evaluated by Rhodamine-phalloidin. The phosphorylated levels of p38 were determined by Western blot. Cells were then treated with p38inhibitor (SB203580), or DMSO, followed by incubation with serum from septic shock patients treated with conventional therapy. Endothelial permeability and F-actin rearrangements were also evaluated as noted above.

RESULTSSerum from CT group increased endothelial permeability, F-actin rearrangements, and phosphorylated levels of p38, which were inhibited by CVVH-ULI treatment. Moreover, in CT group, the serum-induced endothelial hyperpermeability and F-actin rearrangements were inhibited by SB203580, the inhibitor of p38.

CONCLUSIONCVVH combined with ulinastatin decreases endothelial hyperpermeability induced by septic shock through inhibiting p38 MAPK pathways.

Actins ; metabolism ; Cells, Cultured ; Glycoproteins ; therapeutic use ; Hemofiltration ; methods ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Imidazoles ; MAP Kinase Signaling System ; Pyridines ; Shock, Septic ; therapy ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective:To evaluate the efficacy and safety of ultrasound-guided high intensity focused ultrasound(HIFU)on pain intensity,pain sensation and overall survival in patients with advanced pancreatic cancer. Methods:Clinical data of advanced pancreatic cancer patients treated by HIFU were collected from the patients enrolled during August 2020 to September 2022 at the second department for oncology of Yueyang Hospital of Integrated Chinese and Western Medicine affiliated to Shanghai University of Traditional Chinese Medicine.In this study,SPSS 26.0 software was used for the statistical analysis of NRS score and BPI score.The Kaplan-Meier survival analysis method was applied to calculate the median overall survival(OS)and then the survival curve was drawn.At the same time,the incidence of related adverse reactions during and after HIFU treatment was counted. Results:(1)Among the 45 patients,30 patients received HIFU combined with chemotherapy,and the other 15 patients only received HIFU.(2)Among the 45 patients,32 patients had pain relief after HIFU treatment,and the NRS score kept decreased across 1 week,2 weeks,3 weeks and 1 month after HIFU treatment(P＜0.05).The pain sensation score of BPI scale also decreased correspondingly,and the difference was statistically significant(P＜0.05).(3)The median OS of 45 patients was 11.1 months(95％Cl:9.30-1 2.90),of which 30 patients treated with HIFU combined chemotherapy had a median OS of 12.4 months(95％Cl:9.1 8-15.62),and 15 patients treated with HIFU only had a median OS of 4.6 months(95％Cl:1.11-8.10).(4)No serious adverse events were observed in all patients during and after HIFU treatment.Only 5 patients had asymptomatic mild elevation of blood amylase,and the incidence of mild adverse reactions was 11.1％. Conclusion:HIFU can effectively relieve pain and prolong the median survival time in patients with advanced pancreatic cancer.