ObjectiveTo investigate the role of DEAD-box helicase 3 X-linked （DDX3X） in immune-mediated liver injury （ILI）， and to clarify its mechanism by regulating endoplasmic reticulum stress （ERS）-dependent apoptotic pathway and its association with the clinical progression of hepatitis B. MethodsMice were given injection of concanavalin A （ConA） via the caudal vein to establish a model of ILI， PBS （control group） and different concentrations of ConA were injected into the tail vein of hepatocyte-specific DDX3X-knockout mice （DDX3X^ΔHep and DDX3X-flox mice （DDX3X^fl/fl）， respectively.. The log-rank survival analysis， measurement of the serum levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）， and HE staining of liver tissue were performed to assess liver injury， and qRT-PCR and Western Blot were used to measure the mRNA and protein expression levels of glucose-regulated protein 78 （GRP78）， CCAAT/enhancer-binding protein homologous protein （CHOP）， and DDX3X in liver tissue. Intraperitoneal injection of 4-phenylbutyric acid （4-PBA， 100 mg/kg） was performed to inhibit ERS. Serum samples （n=30） and liver tissue samples （n=6） were collected from healthy controls， chronic hepatitis B （CHB） patients， and hepatitis B virus-associated liver failure （HBV-LF） patients； ELISA was used to measure the serum level of DDX3X， and qRT-PCR/Western Blot was used to analyze the expression of targets in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the control group of mice， the expression of DDX3X in the liver of mice induced by ConA was significantly increased after liver injury （P<0.05）， and hepatocyte-specific DDX3X knockout increased the 72-hour survival rate of mice by 55% （compared with 20% in the DDX3X^fl/fl group）， with significant reductions in the serum levels of ALT and AST （P<0.000 1） and the expression levels of the ERS markers GRP78 and CHOP （P<0.05）. After ERS was inhibited by 4-PBA， there was alleviation of liver injury （with reductions in ALT and AST， P <0.001） and a reduction in DDX3X expression （P<0.01）. The analysis of clinical samples showed that the mRNA and protein expression levels of liver DDX3X in CHB patients and HBV-LF patients were significantly higher than those in healthy controls （all P<0.01）， and there was a significant increase in the serum level of DDX3X in HBV-LF patients （P<0.000 1）. ConclusionDDX3X exacerbates ILI by regulating the ERS-dependent apoptotic pathway （GRP78/CHOP）， and its expression is associated with the progression of hepatitis B. Therefore， it can be used as a potential therapeutic target.

ObjectiveTo investigate the effects of modified Buyang Huanwu Tang on cerebral ischemia-reperfusion injury （CI/RI） in mice via the PTEN-induced putative kinase 1/E3 ubiquitin ligase （PINK1/Parkin） signaling pathway-mediated mitophagy， and to explore the underlying mechanism by which modified Buyang Huanwu Tang improves CI/RI. MethodsSeventy-two male C57BL/6J mice were randomly divided into six groups （n = 12 per group）： Sham-operated group， middle cerebral artery occlusion/reperfusion （MCAO/R） model group， low-， medium-， and high-dose modified Buyang Huanwu Tang groups （8.84， 17.68， 35.36 g·kg^-1·d^-1）， and an aspirin group （13.00 mg·kg^-1·d^-1）. Neurological deficit scores were assessed using the Zea-Longa method. Cerebral infarct volume ratio was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Histopathological changes and neuronal injury in brain tissues were observed using hematoxylin-eosin （HE） staining and Nissl staining. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） assay. Mitochondrial ultrastructure in brain tissue was observed by transmission electron microscopy （TEM）. Serum levels of superoxide dismutase （SOD） and malondialdehyde （MDA） were determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expression levels of PINK1， Parkin， microtubule-associated protein 1 light chain 3B （LC3B， LC3Ⅱ/Ⅰ）， and p62 in brain tissues were detected by real-time quantitative reverse transcription PCR （Real-time PCR） and Western blot， respectively. ResultsCompared with the sham-operated group， the MCAO/R model group showed significantly increased neurological deficit scores and cerebral infarct volume ratios （P<0.01）. Severe cortical injury on the infarct side was observed， characterized by decreased neuronal density， cytoplasmic vacuolation， nuclear pyknosis， a marked reduction in Nissl bodies， dissolution of Nissl bodies in the cytoplasm of some pyramidal neurons， and blurred cellular boundaries. The number of TUNEL-positive cells increased significantly （P<0.01）. Mitochondria exhibited cristae membrane rupture and matrix vacuolation， with rupture of the outer mitochondrial membrane and formation of autophagosomes， the number of which increased significantly. Serum SOD activity decreased significantly （P<0.01）， while MDA content increased significantly （P<0.01）. In infarcted brain tissues of model mice， the relative mRNA expression and protein levels of PINK1， Parkin and LC3B were significantly increased （P<0.05， P<0.01）， whereas p62 mRNA and protein expression were significantly decreased （P<0.05， P<0.01）， showing statistical significance. Compared with the model group， all treatment groups showed significantly decreased neurological deficit scores and cerebral infarct volume ratios （P<0.01）. Neuronal density increased significantly， cytoplasmic vacuolation was alleviated， nuclear morphology tended to be more regular and clearer， Nissl body density increased significantly with reduced dissolution and improved contour clarity. The mitochondrial cristae structure was partially restored， with some mitochondria showing autophagosome encapsulation， and the degree of mitochondrial damage was alleviated. Serum SOD activity increased significantly （P<0.01）， while MDA content decreased significantly. The mRNA and protein expression levels of PINK1， Parkin， and LC3Ⅱ/Ⅰ were significantly increased （P<0.05， P<0.01）， while p62 mRNA and protein expression in the low- and medium-dose modified Buyang Huanwu Tang groups were significantly decreased （P<0.05， P<0.01）， showing statistical significance. ConclusionModified Buyang Huanwu Tang can upregulate the protein expression levels of PINK1， Parkin， and LC3Ⅱ/Ⅰ and downregulate p62 protein expression， suggesting that it may improve CI/RI by regulating the expression of proteins related to the PINK1/Parkin signaling pathway. Regulation of the mitophagy pathway may be one of the mechanisms by which modified Buyang Huanwu Tang alleviates CI/RI in mice.

Objective:To explore the effect of combining intermittent θ pulse stimulation (iTBS) of the cerebellum with lower extremity exoskeleton robot support on the balance and walking function of stroke survivors.Methods:Seventy-five stroke survivors complicated with lower extremity dysfunction were divided into an iTBS group, an exoskeleton group and a combined group, each of 25, according to a random number table. In addition to conventional rehabilitation training, the iTBS group was given cerebellar iTBS combined with traditional walking training, the exoskeleton group received sham cerebellar iTBS combined with walking training assisted by a lower extremity exoskeleton robot. The combined group received both therapies. The schedule was once a day, 5 days a week for 3 weeks. Before and after the treatment, the 10-metre walking test (10MWT), the Berg Balance Scale (BBS) and the Fugl-Meyer lower extremity assessment (FMA-LE) were used to evaluate the subjects′ walking ability, balance and lower extremity motor ability. Gait and neuro-electrophysiological tests were also conducted in all three groups.Results:After the treatment, a significant improvement was observed in the 10MWT times, BBS scores, FMA-LE scores, stride frequency and stride speed of all three groups compared with before the treatment. On average, the results of the exoskeleton and combined groups were significantly better than those of the iTBS group, and those of the combined group were significantly better than among the exoskeleton group. Almost everyone′s MEP latency and amplitude had improved significantly compared with before the treatment, but the improvements in the exoskeleton group tended to be superior to those in the iTBS group ( P≤0.05). The latency in the combined group averaged (21.25±1.70)ms, and the amplitude averaged (184.17±6.54)μV, both significantly better than the exoskeleton group′s averages. Conclusions:Cerebellum iTBS combined with lower extremity exoskeleton walker training can significantly improve the motor functioning, balance and walking ability of stroke survivors.

Objective:To explore the effect of combining repeated peripheral (rPMS) and central transcranial magnetic stimulation (rTMS) in treating upper limb motor dysfunction after a stroke.Methods:Seventy-eight patients with upper limb motor dysfunction after a stroke were randomly divided into a control group, an rTMS group and a combined magnetic stimulation group, each of 26. All three groups underwent routine rehabilitation, while the rTMS group was repeatedly given low frequency transcranial magnetic stimulation of the M1 region on the unaffected side, and the combined group also received repeated peripheral magnetic stimulation at Erb′s point on the affected upper limb. There was one treatment session a day, 5 days a week for 3 weeks. Before and after the treatment, everyone′s upper limb motor function was quantified using the Fugl-Meyer upper extremity assessment (FMA-UE) and the Wolf motor function test (WMFT). Skill in the activities of daily living was quantified in terms of a Barthel index (BI). Motor recovery of the upper limbs and hands was assessed using Brunnstrom staging. The latency and amplitude of the motor evoked potentials (MEPs) in the subjects′ affected abductor pollicis brevis muscles were also recorded before and after the treatment. Pearson correlation coefficients quantified the correlation between the changes in FMA-UE scores and MEP amplitudes before and after the treatment in the three groups.Results:There were no significant differences among the three groups before the treatment. Afterward, however, the average FMA-UE, WMFT and BI scores, as well as the upper limb and hand Brunnstrom stages and the average MEP latencies and amplitudes of all the three groups had improved significantly. The combined group′s average results were then significantly better than the other two groups′ averages, except for the upper limb Brunnstrom stages. The increases in MEP amplitude were positively correlated with the increases in FMA-UE scores among the rTMS and the combined group, but there was no significant correlation between them in the control group.Conclusions:The combined application of rPMS and contralateral low frequency rTMS can effectively relieve motor dysfunction in the upper limbs in the early stages after a stroke.

OBJECTIVE To explore the expression levels of microribonucleic acid(miR)-29b and soluble Klotho protein in peripheral blood of the chronic obstructive pulmonary disease(COPD)patients complicated with pulmo-nary infection and analyze the clinical significance.METHODS The clinical data were retrospectively collected from 139 patients with COPD who were treated in Ningxia Medical University General Hospital from Feb.2022 to Feb.2024.The patients were divided into the infection group with 41 cases and the no infection group with 98 cases ac-cording to the status of pulmonary infection.The distribution of pathogens isolated from the patients of the infec-tion group was observed.The levels of serum miR-29b and Klotho were compared between the infection group and the no infection group or of the infection group before and after the anti-infection treatment.The efficiencies of ser-um miR-29b and Klotho in prediction of pulmonary infection were analyzed by means of receiver operating charac-teristic(ROC)curves.RESULTS Totally 46 strains of pathogens were isolated from the infection group,67.39％of which were gram-negative bacteria,26.09％were gram-positive bacteria,and 6.52％were fungi.There were significant differences in the levels of serum miR-29b and Klotho between the infection group and the no infection group(P＜0.05);the expression level of serum miR-29b of the patients in the infection group who were effective for the anti-infection treatment was lower after the treatment than before the treatment,while the expression level of Klotho was higher after the treatment than before the treatment(P＜0.05);the expression level of miR-29b of the patients effective for the treatment was lower than that of the patients ineffective for the treatment,while the expression level of Klotho of the patients effective for the treatment was higher than that of the patients ineffective for the treatment(P＜0.05).ROC curve analysis showed that the area under the curve(AUC)of the joint detec-tion of serum miR-29b and Klotho was 0.927 in prediction of pulmonary infection in the COPD patients,higher than 0.837 and 0.852 of the single detection.CONCLUSIONS The COPD patients complicated with pulmonary in-fection show the high expression level of serum miR-29b and low expression level of Klotho.The joint detection of the two indexes has high efficiency in prediction of pulmonary infection in the COPD patients,which can be used as reliable indexes for assessment of the curative effect after the anti-infection treatment.

Objective To explore the application value of serum heat shock protein 90α(Hsp90α)combined with β2-microglobulin(β2-MG)detection in the early diagnosis and prognosis of colorectal cancer(CRC).Methods A total of 101 patients with CRC who received their first treatment were selected as the CRC group(no surgery,radiotherapy or chemotherapy before enrollment),100 patients with benign colon tumor admitted to our hospital during the same period were selected as the benign tumor group and 94 healthy individuals who underwent physical examinations in our hospital were used as the control group.The levels of serum Hsp90α and β2-MG in all subjects were detected by ELISA.Follow-up was conducted every three months within the first two years after surgery,and every six months in the third year,for a total follow-up period of 3 years,and the survival status of CRC patients within 3 years was recorded.ROC curve analysis was used to evaluate the diagnostic value of serum Hsp90α and β2-MG for CRC.The Kaplan-Meier method was used to draw the survival curves of CRC patients with different expression levels of Hsp90α and β2-MG.Results The serum levels of Hsp90α and β2-MG increased successively in the control group,the benign tumor group and the CRC group(P＜0.05).There were no significant differences in serum levels of Hsp90α and β2-MG between CRC patients of different genders,ages and lesion locations.The levels of serum Hsp90α and β2-MG in patients with tumor diameter＞5 cm,low differentiation degree,TNM stage Ⅲ and lymph node metastasis were higher than those in patients with tumor diameter≤5 cm,moderate to high differentiation degree,TNM stage Ⅰ-Ⅱ and no lymph node metastasis(P＜0.05).The ROC results showed that the combined diagnostic efficacy of serum Hsp90α and β2-MG was superior to that of serum Hsp90α or β2-MG alone(Z=2.433,2.435,P＜0.05).Based on the average expression levels of serum Hsp90α and β2-MG in CRC patients(76.66 μg/L and 6.52 μg/L),patients were divided into the high expression group of Hsp90α(59 cases),the low expression group of Hsp90α(42 cases),the high expression group of β2-MG(63 cases)and the low expression group of β2-MG(38 cases).Among the 101 CRC patients,31 died and 70 survived,with a total survival rate of 69.31％.The Kaplan-Meier survival curve showed that the 3-year survival rates of patients with high expression levels of Hsp90α and β2-MG were lower than theose of patients with low expression levels of Hsp90α and β2-MG(P＜0.01).Conclusion The expression levels of serum Hsp90α and β2-MG are both elevated in CRC patients,and they are closely related to the occurrence,development and prognosis of CRC.

Objective To systematically evaluate the effect of hysteroscopic uterine septum resection on the pregnancy live birth rate in patients with septate uterus.Methods The randomized controlled studies and cohort studies on the impact of hysteroscopic uterine septum resection on the pregnancy live birth rate in the patients with septate uterus were retrieved from PubMed,Web of Science,CNCN.cn,and Wanfang databases.The retrieval time was from the establishment of the database to March 2024.The NOS scale was used to con-duct the literature quality evaluation.The Statal0.0 statistical software was used for conducting the meta a-nalysis,and the main evaluation index was the pregnancy live birth rate.Results A total of 9 articles on the treatment of septate uterine were included,involving 821 subjects,including 528 septate uterine patients with hysteroscopic uterine septum resection and 293 septate uterine patients with conservative treatment.The meta analysis showed that the live birth rate in the patients with septate uterine undergoing hysteroscopic uterine septum resection was higher than that in the patients with septate uterine undergoing the conservative treat-ment(I2=38.0％,OR=1.60,95％CI:1.05-2.44,P=0.028).Conclusion Hysteroscopic uterine septum resection could improve the pregnancy outcome and increase the live birth rate.It is a minimally invasive sur-gery with good effect and high safety,which has good clinical application prospects and is worth popularizing.

Objective To investigate the mechanism of action of the nuclear factor erythroid 2-related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)signaling pathway in non-alcoholic fatty liver disease(NAFLD),and to explore the mechanism of Gegen QinLian Decoction for the treatment of NAFLD,using in vivo and in vitro experiments.Methods Rats were fed with high-fat chow for 24 weeks to induce NAFLD,and were then divided randomly into normal(C),model(M),high-,medium-,and low-dose Gegen QinLian Decoction(GGQLT-H,GGQLT-M,GGQLT-L),and metformin(Met)groups.From week 25 onwards,the rats were administered the corresponding drugs by gavage for 2 weeks according to the grouping,until sampling.Levels of the oxidative stress markers malondialdehyde(MDA)and glutathione(GSH)in the liver tissues were measured in each group using biochemical kits and ferrous iron(Fe2+)in rat liver tissues was detected using a Fe2+kit.Nrf2,heme oxygenase-1(HO-1),SLC7A11,glutathione synthetase(GSS),GPX4,and acyl coenzyme A synthetase 4(ACSL4)mRNA levels in rat liver tissues were measured by reverse transcription quantitative polymerase chain reaction.For cellular experiments lipid acc umulation was induced in HepG2 hepatocellular carcinoma cells using 1 mmol/L free fatty acid,to mimic the NAFLD in vitro model.Different concentrations of Gegen QinLian Decoction and metformin-containing serum were added for treatment.Lipid accumulation was detected in the cells in each group by Oil red O staining.The MDA and GSH contents of HepG2 cells in the different groups were determined using appropriate kits,and the ferrous contents were detected using a cell-specific ferrous kit.Expression levels of Nrf2,HO-1,SLC7A11,GSS,GPX4,and ACSL4 mRNA was detected in each group of cells using reverse transcription quantitative polymerase chain reaction.Results In the animal experiments,MDA and Fe2+liver levels were significantly higher in the M group than in the C group,while GSH levels were significantly lower(P＜0.01).GGQLT-H,GGQLT-M and Met groups showed significantly reduced MDA and Fe2+and elevated GSH levels compared with the M group(P＜0.01,P＜0.05).High-and medium-dose Gegen QinLian Decoction and metformin increased Nrf2,HO-1,GSS,and GPX4 mRNA and decreased ACSL4 mRNA expression levels(P＜0.01,P＜0.05).In cellular experiments,lipid droplets were significantly increased in the HepG2 cell M group compared with those in the C group,and lipid droplets were significantly reduced by Gegen QinLian Decoction and metformin.MDA and Fe2+levels were significantly increased and GSH levels were significantly decreased in the HepG2 M group compared with the levels in the C group(P＜0.01),while all doses of Gegen QinLian Decoction and metformin significantly decreased MDA and Fe2+levels(P＜0.01)and increased the GSH content(P＜0.01,P＜0.05).Nrf2,GSS,GPX4,and SLC7A11 mRNA expression levels in the GGQLT-H group,Nrf2,HO-1,and SLC7A11 in the GGQLT-L group,HO-1,SLC7A11,and GSS in the GGQLT-M group,and GSS,Nrf2,and HO-1 in the Met group were all significantly increased compared with the findings in the M group(P＜0.01,P＜0.05).ACSL4 mRNA expression levels were significantly decreased in the GGQLT-M and GGQLT-L groups and the Met group(P＜0.01,P＜0.05).Conclusions Gegen QinLian Decoction can improve NAFLD by inhibiting ferroptosis,and its mechanism may he related to regulation of the Nrf2/SLC7A 11/GPX4 signaling pathway.