Objective Uranium compounds are critical materials in the nuclear industry, and their mining, processing, and use pose occupational exposure risks. Epidemiological studies have shown that uranium exposure can impair health, with acute uranium poisoning primarily causing structural and functional damage to the kidneys. Currently, the molecular mechanisms underlying uranium-induced kidney injury remain unclear, and traditional biological models used in toxicological research are inadequate for simulating the human microenvironment. This study employed a human kidney organoid model system to elucidate the nephrotoxic mechanisms of uranyl ions, providing a scientific basis for the prevention and control of uranium poisoning. Methods Kidney organoids were constructed using the human embryonic stem cell line H1 and exposed to solutions with different uranyl ion concentrations. Morphological observation, ATP detection, reactive oxygen species detection, apoptosis assay, and untargeted metabolomics analysis were performed. Mechanisms of toxicity were further explored through KEGG pathway enrichment analysis. Results Uranium exposure led to structural damage in the organoids, accompanied by a dose-dependent decrease in ATP levels, accumulation of reactive oxygen species, and increased apoptosis rate. After 24-hour exposure to 300 μmol/L uranium, significantly disturbed differential metabolites and five core metabolic pathways were identified. Conclusion Uranium induces oxidative stress, mitochondrial dysfunction (ATP reduction), and metabolic disorder (disruption of phospholipid/amino acid metabolism), which synergistically cause DNA damage, apoptosis, and/or necrosis. These findings provide new insights into the mechanisms of uranium-induced kidney injury and support the development of prevention and control strategies.

Objective:To investigate the effects of ginsenoside Rg1 (GS-Rg1) on the secretion of exosomes (MSC-Exo) and expression of angiogenesis related microRNAs (miRNAs) in mesenchymal stem cells.Methods:Human umbilical cord blood mesenchymal stem cells (hUCBMSCs) were divided into experimental group and control group. The experimental group was treated with GS-RG1 at a final concentration of 40 mg/L, while the control group was treated with phosphate buffered saline (PBS) at the same volume. Both groups were cultured for 24 h. The morphology of MSC-Exo was observed by transmission electron microscopy; the characteristic surface markers were identified by Western blot; the concentration of MSC-Exo was detected by dicootanobutyric acid protein quantification method, and the expression of 8 miRNAs related to angiogenesis in MSC-Exo was detected by reverse transcription polymerase chain reaction (RT-PCR).Results:After 24 h of incubation, MSC-Exo with a circular membrane vesicle structure was visible. MSC-Exo was positive for the expression of the characteristic surface markers CD9, CD63 and TSG101. After 24 h of intervention, the concentration of MSC-Exo protein were (1.080±0.019)μg/μl and (0.881±0.032)μg/μl in the experimental group and control group, respectively, with statistically significant difference ( P<0.01). The expression of miR-126-3p, miR-21, miR-146a-5p and miR-125b-5p in the GS-Rg1 group were significantly higher than that in the control group, while the expression of miR-16-5p was significantly lower than that in the control group (all P<0.05). Conclusions:GS-Rg1 promotes the secretion of MSC-Exo and enhances the expression of angiogenesis-related miRNAs within Exo to promote angiogenesis.

Objective To discuss the clinical efficacy of surgery for chronic subdural hematoma assisted by rigid neuroendoscope and its surgical techniques. Methods Clinical data of 161 patients with chronic subdural hematoma from August 2009 to December 2015 was analyzed retrospectively. 74 of them experienced surgeries assisted by rigid neuroendoscope (endoscope group) and other 87 cases were operated without neuroendoscope (routine group) during the same period. Results Although there were significant difference in operative duration between the two groups, complications, ratio of total removal of hematoma after surgery, postoperative inpatient duration and recurrent rate of hematoma were more advantageous in endoscope group. The operative duration of endoscope group with (112.68 ± 34.86) min was longer than that of routine group with (74.11 ± 28.23) min (t = 7.75, P = 0.000), while the postoperative inpatient duration of endoscope group with (8.23 ± 2.01) d was shorter than that of another group with (10.79 ± 5.02) d (t = -4.12, P = 0.000). There were no surgical associated complications in endoscope group, but 1 patient in routine group experienced intracerebral hematoma of frontal lobe and associated aphemia. Total removal of hematoma was confirmed in endoscope group with 98.65% (73/74), which was higher than that in routine group with 86.21% (75/78) (χ2 = 8.34, P = 0.004). Hematoma recurrence was found in 16 cases of routine group (18.39%), but more superiority in endoscope group with 1.35% (χ2 = 12.29, P = 0.000). Outpatient follow-up was carried out in all patients from 6 to 38 months with an average duration of 30.06 months. In 17 cases with recurrent hematoma during follow-up, 15 of them were cured by a second surgery, and another 2 patients were cured by atorvastatin. Conclusion As a simple, safe and effective technique, the application of rigid neuroendoscope during surgery for chronic subdural hematoma is more advantage than routine surgery. A self-made suction with adjustable soft curved tip is suitable for such procedure.

Objective:To observe the effect of old RBCs transfusion on cognitive function in rats and the improvement effect of mi -nocycline.Methods: Male SD rats at the age of 6 months were randomly divided into 4 groups.The RBCs were obtained from male rats by centrifuging the total blood and stored at 4℃.The rats of fresh RBCs group (group F) were transfused with the RBCs stored for 1 day.The rats of old RBCs group (group O) were transfused with the RBCs stored for 7 days.The rats of treatment group (group T) received 40 mg· kg-1 minocycline with intraperitoneal injection before the transfusion .The rats of the control group ( group C) were transfused with the normal saline .The brain levels of IL-1βand IL-6 were determined with Quantikine ELISA kits in 24 hours after the blood transfusion (n=6).The rats were subjected to Barnes maze tests after 1 week of the blood transfusion (n=10).Results:The brain levels of IL-1βand IL-6 in group O were higher than those in group C and F (P<0.05), which were lower in group T than those in group O(P<0.05).The rats of group O spent longer time finding the target box than those of group C and F in the Barnes maze (P<0.05), and the time was shorter in group T than that in group O (P<0.05).Conclusion: Old RBCs transfusion plays a role in neuro-inflammation and induces cognitive dysfunction in rats , which may be improved by minocycline .

Objective To evaluate the effect of lithium chloride (LiCl) pretreatment on isoflurane-induced cognitive dysfunction and inflammatory response in hippocampus in aged rats.Methods Eighty 20-month-old male Sprague-Dawley rats,weighing 350-400 g,were randomly assigned into 4 groups (n =20 each):control group (group C),1.4％ isoflurane group (group I),100 mg/kg LiCI + 1.4％ isoflurane group (group L+ I),and 100 mg/kg LiC1 group (group L).Group I was exposed to 1.4％ isoflurane in 30％ O2-70％ N2 for 6 h,while group C was exposed to 30％ O2-70％ N2 only.LiCl 100 mg/kg was injected intraperitoneally once a day for 3 consecutive days and isoflurane anesthesia was performed on 4th day in group L + I.LiCl 100 mg/kg was injected intraperitoneally once a day for 3 consecutive days and then the rats inhaled 30％ O2-70％ N2 for 6 h on 4th day in group L.Blood samples were taken immediately after the end of anesthesia for blood gas analysis.Hippocampi were isolated 24 h after the end of anesthesia for determination of the expression of glycogen synthase kinase-3β (GSK-3β) and acetyl-NF-κB (Lys310) (by Western blot) and TNF-α,IL-1β and IL-6 mRNA (by RT-PCR).The levels of TNF-α,IL-1β and IL-6 were determined by ELISA and the contents of TNF-α,IL-1β and IL-6 were calculated.The cognitive function was assessed on 2nd day after the end of anesthesia.Results Compared with group C,the expression of GSK-3β and acetyl-NF-κB (Lys310) was significantly up-regulated,the expression of TNF-α,IL-1β and IL-6 mRNA and contents of TNF-α,IL-1β and IL-6 were increased,the escape latency was prolonged,and the time of staying at the original platform quadrant was shortened in group I (P ＜ 0.05).Compared with group I,the expression of GSK-3β and acetyl-NF-κB (Lys310) was significantly down-regulated,the expression of TNF-α,IL-1β and IL-6 mRNA and contents of TNF-α,IL-1β and IL-6 were decreased,the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in group L + I (P ＜ 0.05).Conclusion LiC1 pretreatment can improve isoflurane-induced cognitive dysfunction and inhibition of inflammatory response in hippocampus is involved in the mechanism in aged rats.

Objective To investigate the effects of isoflurane anenthesia on myocyte enhancer factor 2(MEF2) signaling pathway in neonatal rat hippocampus. Methods Twenty-four 5-day-old SD rats of both sexes,weighing 10-13 g, were randomly divided into 2 groups ( n = 12 each): control group (group C) and isoflurane group (group I). In group I, 1.5% isoflurane in 100% O2 was inhaled for 6 h. Group C received no treatment.Three rata in each group were sacrificed at 2, 4, 6 h of isoflurane anenthesia and 24 h after isoflurane anenthesia (T1-4), and the hippocampi removed for determination of MEF2 mRNA, synGAP Ⅰ mRNA, Arc mRNA and synapsinⅠ mRNA expression (by PT-PCR) and synapsin Ⅰ protein expression (by Western blot).Results Compared with group C, the expression of MEF2 mRNA, synGAP Ⅰ mRNA, Arc mRNA and synapsin Ⅰ mRNA at T1-3 and synapsin Ⅰ protein at T2-4 was up-regulated in group I ( P ＜ 0.05). Conclusion Inhalation of anaesthetic concentration of isoflurane may affect synapse formation during the development of central nervous system by actirating hippocampal MEF2 signaling pathways in neonatal rats.

This study examined the effects of clinically relevant concentrations of isoflurane on the amplitude of NMDA receptor current (I(NMDA)) and the expression of cytochrome C in cultured developing rat hippocampal neurons. The hippocampi were dissected from newborn Sprague-Dawley rats. Hippocampal neurons were primarily cultured for 5 days and then treated with different concentrations of isoflurane [(0.25, 0.5, 0.75, 1 minimum alveolar concentration (MAC))]. The peak of I(NMDA) was recorded by means of the whole cell patch clamp technique. The cytochrome C level was detected by Western blotting and quantitative real-time PCR. Our results showed that isoflurane (0.25, 0.5, 0.75 and 1 MAC) potentiated the amplitude of I(NMDA) by (116±8.8)%, (122±11.7)%, (135±14.3)% and (132±14.6)%, respectively, and isoflurane increased the mRNA expression of cytochrome C in a concentration-dependent manner. The cytochrome C mRNA expression reached a maximum after 0.5 MAC isoflurane stimulation for 6 h (P<0.05). It was concluded that isoflurane enhances the expression of cytochrome C in cultured rat hippocampal neurons, which may be mediated by facilitation of NMDA receptor.

This study examined the effects of clinically relevant concentrations of isoflurane on the amplitude of NMDA receptor current (INMDA) and the expression of cytochrome C in cultured developing rat hippocampal neurons.The hippocampi were dissected from newborn Sprague-Dawley rats.Hippocampal neurons were primarily cultured for 5 days and then treated with different concentrations of isoflurane [(0.25,0.5,0.75,1 minimum alveolar concentration (MAC))].The peak of INMDA was recorded by means of the whole cell patch clamp technique.The cytochrome C level was detected by Western blotting and quantitative real-time PCR.Our results showed that isoflurane (0.25,0.5,0.75 and 1 MAC) potentiated the amplitude of INMDA by (116±8.8)％,(122±11.7)％,(135±14.3)％ and (132±14.6)％,respectively,and isoflurane increased the mRNA expression of cytochrome C in a concentration-dependent manner.The cytochrome C mRNA expression reached a maximum after 0.5 MAC isoflurane stimulation for 6 h (P＜0.05).It was concluded that isoflurane enhances the expression of cytochrome C in cultured rat hippocampal neurons,which may be mediated by facilitation of NMDA receptor.

Objective To investigate the effect of ketamine on cAMP response element binding protein pbosphorylation(p-CREB)in hippocampus of neonatal rats.Methods Seventy-five 7-day old SD rats of both sexes were randomly divided into 3 groups(n=25 each):control group(group C)and 2 ketamlne groups(group K1,K2)which received 7 subcutaneous injections of ketamine 10 and 20 mg/kg respectively at 90 min intervals.The animsla were decapitated at 24 h after fwst ketamine injection.The brains were immediately removed and the hippocampi were isolated for detection of neuronal apoptosis by TUNEL.Apoptosis index wag calculated.The expression of p-CREB Wag meagured by immuno-histochemistry and the expression of BDNF mRNA and Bcl-2 mRNA was detected by RT-PCR.Cognitive function Wag agsessed using Morris water maze test at 6 weeks after first ketamine injection.Results The apoptosis index Wag significantly increased while the expression of CREB,BDNF mRNA and Bcl-2 mRNA was down-regulated in group K1 and K2 as compared with group C.The apoptosis index Wag significantly higher and the expression of p-CREB and BDNF mRNA and Bcl-2 mRNA Wag significantly lower in group K2 than in group K1.The latent period of escape was significantly longer in group K2 than in group C and K1.Conclusion Ketamine 20 mg/kg administered in neonatal rats can decrease cognitive function when they grow up by increasing neuronal apoptosis induced by down-regulatlon of the expression of p-CREB,BDNF and Bcl-2.

AIM:To construct recombinant adenovirus vector carrying human miR-133a and study its expression in human mesenchymal stem cells(hMSCs).METHODS:The PCR product containing miR-133a was amplified from human genomic DNA and inserted into the adenoviral shuttle vector pAdTrack-CMV.Then the recombinant shuttle plasmid linearized by pmeⅠwas cotransformed into competent E.coli.BJ5183 with the adenoviral backbone plasmid pAdEasy-1 to generate the recombinant adenovirus vector rAd-mir-133a.rAd-mir-133a was then packaged and amplified in human embryonic kidney 293(HEK293) cells.The purified rAd-miR-133a was used to infect the hMSCs and the expression of miR-133a was detected by non-quantitative RT-PCR and real-time PCR.RESULTS:The recombinant adenovirus shuttle vector pAdTrack-CMV-miR-133a was constructed and verified by restriction endonuclease analysis and DNA sequence analysis.rAd-miR-133a was successfully packaged and amplified in HEK293 cells.The transcriptions of primary miR-133a and mature miR-133a were over-expressed in the hMSCs infected with rAd-miR-133a.CONCLUSION:The recombinant adenovirus vector carrying human miR-133a is successfully constructed,which lay a foundation for miR-133a function study.