Objective:To observe the effect of robot-assisted gait training (RAGT) supplemented with intermittent theta burst stimulation (iTBS) of the cerebellum in restoring lower limb function after anterior cruciate ligament reconstruction (ACLR).Methods:Eighty ACLR patients were randomly divided into a control group, a magnetic stimulation group, a robot group and a combined group, each with 20 members. The robot and magnetic stimulation groups underwent RAGT and cerebellar iTBS before conventional training, while the combined group received iTBS followed by RAGT and then conventional training. The treatments were administered once a day, three days per week for four weeks. Before and after the intervention, the peak torque ratio of the knee flexors and extensors (H/Q), peak torque of the knee extensors (PT), and knee repositioning angle difference were measured. Knee function and balance (using the Berg Balance Scale (BBS)) were also assessed.Results:The combined group demonstrated significantly better quadriceps PT and H/Q% than the other 3 groups. Knee repositioning angle difference improved significantly in all of the groups after the treatment, with the combined group showing the smallest difference (5.00±1.21)°, significantly better than the other three groups. Lysholm and BBS scores had also improved significantly in all of the groups, with the combined group′s improvements again significantly better than those of the other groups.Conclusion:Intermittent theta burst stimulation of the cerebellum combined with robot-assisted gait training can significantly improve knee function and balance after ACLR.

Background and aims:Primary sclerosing cholangitis(PSC)is an autoimmune liver disease characterized by complex pathogenesis and limited available therapeutic options.The mechanisms underlying the development and progression of PSCs remain unclear.Liver X receptor beta(LXR-β)is recognized to modulate lipid metabolism and immune response,but its specific involvement in the PSC has not been elucidated.Here,we explored the role and mechanism of LXR-β in PSC induced by 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-collidine(DDC).Methods:CRISPR-Cas9 technology was applied to generate Abcb4(coding MDR2,next named as Mdr2),Nr1h2(coding LXR-β,next named as Lxrβ),and Rag2(coding RAG2)knockout mice.DDC was used to induce PSC.Hematoxylin and eosin and Sirius red staining were used to assess the extent of hepatic injury and fibrosis.Flow cytometry was used to observe immune cell subsets.Results:We observed a declining trend in hepatic Lxrβ in the PSC model.Unexpectedly,Lxrβ knockout failed to modulate DDC-induced PSC pathogenesis.Concomitantly,assessment of the influence of Rag2 deficiency on PSC progression revealed the absence of aggravated or alleviated hepatic injury or fibrosis in the Rag2-/-DDC mice.However,Lxrβ depletion intensified DDC-induced PSC in the Rag2-/-mice,with more abundant infiltrative inflammatory cells and more severe liver fibrosis.Compared with Rag2-/-DDC mice,Lxrβ-/-Rag2-/-DDC mice had higher serum ALT and AST levels and mRNA expression of proinflammatory and profibrotic genes.Flow cytometry showed that LXR-β deficiency resulted in a diminished population of hepatic innate immune cells.Conclusion:This study indicated innate immune cell LXR-β deficiency can exacerbate hepatic injury and fibrosis in murine models of PSC suggesting that LXR-β may regulate the function of innate immunity in the fibrotic advancement of PSC.

BackgroundThe functional changes of the default mode network （DMN） are closely related to the onset of major depressive disorders. However， the relationship between the DMN subsystem （core subsystem， dorsomedial prefrontal cortex subsystem， medial temporal lobe subsystem） and symptoms of first-episode major depressive disorder remains unclear. ObjectiveTo investigate abnormal functional connectivity between DMN subsystems and the whole brain in first-episode major depressive disorder patients during the resting-state， and to analyse the correlations between these functional connectivity patterns and clinical symptoms， so as to reveal the potential neural mechanisms from the perspective of DMN subsystem. MethodsFrom September 2020 to September 2023， a total of 64 first-episode outpatients and inpatients meeting the diagnostic criteria for major depressive disorder in the Diagnostic and Statistical Manual of Mental Disorders， fourth edition （DSM-IV） were enrolled at the Inner Mongolia Autonomous Region Mental Health Center as the study group. During the same period， 54 healthy volunteers matched for age， gender， and years of education were recruited from the community as the control group. Both groups were assessed using the Hamilton Depression Scale-24 item （HAMD-24）. Resting-state functional magnetic resonance images （rs-fMRI） of the two groups were acquired using a Siemens 3.0 T scanner， and differences in functional connectivity between DMN subsystems （core subsystem， dorsomedial prefrontal cortex subsystem， medial temporal lobe subsystem） and the whole brain were compared. The functional connectivity values of brain regions with statistically significant differences between the two groups were extracted. Spearman's rank correlation coefficient analysis was used to investigate the correlation between these functional connectivity values and HAMD-24 scores of the study group. ResultsUltimately， 46 patients and 43 controls completed the study. Compared with the control group， the study group exhibited significantly stronger functional connectivity in the following pathways： between the right superior parietal lobule （core subsystem） and right cerebellar lobule VIII （t=3.954， P<0.05， GRF-corrected）， between the right lateral temporal cortex （dorsomedial prefrontal cortex subsystem） and right cerebellar lobule VIII， right and left hippocampi， right medial， and paracingulate gyrus （t=4.595， 4.208， 5.200， 4.038， P<0.05， GRF-corrected）， and between the temporoparietal junction （dorsomedial prefrontal cortex subsystem） and left lingual gyrus and right cerebellar lobule VIII （t=3.557， 4.274， P<0.05， GRF-corrected）. Conversely， weaker functional connectivity was observed between the right inferior frontal gyrus and left gyrus rectus （t=-3.824， P<0.05， GRF-corrected）. Furthermore， within the study group， the functional connectivity values between the right lateral temporal cortex and right hippocampus， as well as between the temporoparietal junction and right cerebellar lobule VIII， were both negatively correlated with the HAMD-24 cognitive impairment factor score （r=-0.306， -0.318， P<0.05）. ConclusionIncreased functional connectivity between the DMN （specifically its core and dorsomedial prefrontal cortex subsystems） and cerebellum， partial limbic system， and lingual gyrus may be associated with the neuropathology of first-episode major depressive disorder. Furthermore， alterations in functional connectivity between the dorsomedial prefrontal cortex subsystem and both the cerebellum and hippocampus in these patients may be related to cognitive function. ［Funded by 2019 Annual Inner Mongolia Autonomous Region Natural Science Foundation Project （number， 2019MS03038）； 2023 Annual Inner Mongolia Autonomous Region Natural Science Foundation Project （number， 2023MS08028）］

Objective The analgesic effect of manual acupuncture on acute adjuvant arthritis(AA)rats was evaluated using flurbiprofen cataplasm as a positive control,and the role of mast cells in the mechanism of analgesia was explored.Methods 24 SD rats were randomly divided into model group,10-minute manual acupuncture group,and 30-minute flurbiprofen cataplasm treatment group.AA rat models were established,and treatments were applied at the Zusanli acupoint,while the model group received no treatment.The rats'pain thresholds under mechanical and thermal stimuli were measured before and after the therapy.Acupoint tissue sections were collected and stained,and the mast cell degranulation rate at the acupoint tissue was calculated for each experimental group.Results Mechanical and thermal pain thresholds were significantly increased in 10-minute manual acupuncture group compared to those before therapy(P＜0.000 1),while there was no significant difference in mechanical and thermal pain pain threshold recovery rates between 10-minute manual acupuncture group and 30-minute flurbiprofen cataplasm treatment group(P＞0.05).The mast cell degranulation rate in 10-minute manual acupuncture group and the 30-minute flurbiprofen cataplasm treatment group was significantly higher than that of the model group(P＜0.001).Conclusions Short-term application of manual acupuncture provides immediate analgesia in AA rats,comparable to flurbiprofen cataplasm treatment.The analgesic effects of manual acupuncture and flurbiprofen cataplasm treatment may be closely related to the degranulation of mast cells in the Zusanli acupoint tissue.This study provides an optimized clinical protocol for treating inflammatory joint diseases while laying the groundwork for future research on treatment mechanisms,long-term outcomes,and combination therapy applicability in varied patient groups.

Objective The analgesic effect of manual acupuncture on acute adjuvant arthritis(AA)rats was evaluated using flurbiprofen cataplasm as a positive control,and the role of mast cells in the mechanism of analgesia was explored.Methods 24 SD rats were randomly divided into model group,10-minute manual acupuncture group,and 30-minute flurbiprofen cataplasm treatment group.AA rat models were established,and treatments were applied at the Zusanli acupoint,while the model group received no treatment.The rats'pain thresholds under mechanical and thermal stimuli were measured before and after the therapy.Acupoint tissue sections were collected and stained,and the mast cell degranulation rate at the acupoint tissue was calculated for each experimental group.Results Mechanical and thermal pain thresholds were significantly increased in 10-minute manual acupuncture group compared to those before therapy(P＜0.000 1),while there was no significant difference in mechanical and thermal pain pain threshold recovery rates between 10-minute manual acupuncture group and 30-minute flurbiprofen cataplasm treatment group(P＞0.05).The mast cell degranulation rate in 10-minute manual acupuncture group and the 30-minute flurbiprofen cataplasm treatment group was significantly higher than that of the model group(P＜0.001).Conclusions Short-term application of manual acupuncture provides immediate analgesia in AA rats,comparable to flurbiprofen cataplasm treatment.The analgesic effects of manual acupuncture and flurbiprofen cataplasm treatment may be closely related to the degranulation of mast cells in the Zusanli acupoint tissue.This study provides an optimized clinical protocol for treating inflammatory joint diseases while laying the groundwork for future research on treatment mechanisms,long-term outcomes,and combination therapy applicability in varied patient groups.

Objective:To observe the effect of robot-assisted gait training (RAGT) supplemented with intermittent theta burst stimulation (iTBS) of the cerebellum in restoring lower limb function after anterior cruciate ligament reconstruction (ACLR).Methods:Eighty ACLR patients were randomly divided into a control group, a magnetic stimulation group, a robot group and a combined group, each with 20 members. The robot and magnetic stimulation groups underwent RAGT and cerebellar iTBS before conventional training, while the combined group received iTBS followed by RAGT and then conventional training. The treatments were administered once a day, three days per week for four weeks. Before and after the intervention, the peak torque ratio of the knee flexors and extensors (H/Q), peak torque of the knee extensors (PT), and knee repositioning angle difference were measured. Knee function and balance (using the Berg Balance Scale (BBS)) were also assessed.Results:The combined group demonstrated significantly better quadriceps PT and H/Q% than the other 3 groups. Knee repositioning angle difference improved significantly in all of the groups after the treatment, with the combined group showing the smallest difference (5.00±1.21)°, significantly better than the other three groups. Lysholm and BBS scores had also improved significantly in all of the groups, with the combined group′s improvements again significantly better than those of the other groups.Conclusion:Intermittent theta burst stimulation of the cerebellum combined with robot-assisted gait training can significantly improve knee function and balance after ACLR.

In order to achieve the purpose of rapid detection of duck astrovirus type 1(DAstV-1),specific primers were designed based on the conservative region of ORF1a which belonged to DAstV-1(WF1202 strain).A real-time fluorescent quantitative PCR(qPCR)detective method for DAstV-1 was established.Clinical samples were detected by the qPCR method and the positive samples were used for virus isolation and identification.Results showed that the detection limit of the established method was 4.64×103 copies/μL,which was 10 times higher than the normal RT-PCR method.In addition,no cross-reactions were found with other common infectious disease pathogens in poultry,indicating that the qPCR method had good specificity.What's more,the coef-ficient of variations(Cv)in intra-and inter-assays were 0.85％-2.85％and 0.21％-2.94％,re-spectively,both less than 3％,indicating that the qPCR method had a good repeatability.Using this method,35 tissue samples from different duck farms in 10 provinces from 2020 to 2022 were detected for DAstV-1.Results showed that the positive rate was 25.71％(9/35),and the coinci-dence rate was 94.29％when compared with the normal RT-PCR method.A positive sample ran-domly taken for the virus isolation through duck embryo passage,and the allantoic fluid was col-lected and then was verified by the qPCR method and inoculated with 1-day-old healthy ducklings for the animal regression experiment.The infected ducklings suffered from transient disease but did not die.The liver tissues were all positive with DAstV-1 when detected by qPCR.Meanwhile,autopsy showed that there were slight changes in the livers,and the histopathological observation showed that the liver cells were steatosis.These findings indicated that the isolated DAstV-1 strain had weak pathogenicity and might be a low virulent strain.To sum up,the qPCR detection method of DAstV-1 was successfully established in this work,and could provide technical support for clini-cal diagnosis,isolation and identification,and molecular epidemiology monitoring of DAstV-1.

This study investigates the metabolic regulatory effects of exogenous lactate on obesity mice induced by high-fat diet.We established obesity and metabolic disorder C57 mice model using a synthetic high-fat forage containing 60% fat.Some mice were fed with high-fat diet for 4 weeks to establish the model, being given 500 mg/(kg?d) lactate with ip for 4 weeks at the same time; the others were fed with high-fat diet for 8 weeks to establish the model, being given 500 mg/(kg?d) lactate 4 weeks after 4 weeks of modeling.During the trial period, the change of body weight and food intake, as well as serum glucose, lactate, triglycerides, insulin, and liver glycogen levels of both groups of mice were measured.Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were used to assess glucose metabolism and insulin resistance in the body.At the end of the experiment, adipose tissue was dissected for weighing and histopathological examination.The expression of lipid synthesis and lipolysis genes in adipose tissue was detected by real-time PCR.The results showed that: (1) in the 4-week preventive medication trial, lactate had no significant effect on the body weight of normal and high-fat diet (HFD) mice, yet it increased the subcutaneous fat/visceral fat weight ratio; lactate could significantly reduce fasting blood glucose and liver glycogen levels in HFD mice while increasing blood lactate levels, significantly improving impaired glucose tolerance in HFD mice; lactate could improve the size and arrangement of adipocytes in the HFD group while significantly down-regulating the expression of fatty acid synthesis and lipolysis genes in adipose tissue; (2) in the 8-week treatment, both routes of lactate administration could partially reduce body weight in HFD group mice and reduce food intake, with the improvement trend for fat weight; both routes of lactate administration could significantly reduce fasting blood glucose levels in HFD mice, while significantly improving glucose and insulin tolerance, with some improvement of fasting insulin levels and insulin resistance index; both routes of lactate administration showed different degrees of improvement effect on adipocyte morphology in obese mice while significantly down-regulating lipolysis gene expression in adipose tissue.Therefore, for high-fat diet-induced obese mice with metabolic imbalance, exogenous lactate can stimulate glucose metabolism, inhibit adipose tissue lipolysis, and prevent adipocyte hypertrophy, thereby improving glucose tolerance and insulin sensitivity and reducing sugar-lipid metabolic disorder.

Objective:To investigate the relationship of antinuclear antibody (ANA) status with clinical features and malignancy risk in adult patients with dermatomyositis.Methods:A retrospective analysis was performed to analyze clinical data from 101 inpatients with dermatomyositis in Department of Dermatology, the First Affiliated Hospital of Soochow University from April 2008 to April 2018. These patients were divided into ANA-positive group and ANA-negative group, and differences in myopathy and malignancy risks as well as other clinical features were analyzed between the 2 groups. A 2-year follow-up was undertaken among 92 patients. Chi-square test was used to analyze and compare clinical features between the 2 groups, and a multivariate regression model was used to analyze the relationship of ANA status with amyopathic dermatomyositis and malignancies.Results:Among the 101 patients with dermatomyositis, there were 42 males and 59 females, aged 55.13 ± 14.63 years; 14 patients had amyopathic dermatomyositis, 6 patients had hypomyopathic dermatomyositis, and 81 patients had myopathic dermatomyositis; 42 (41.58%) cases were positive for ANA, and 59 (58.41%) were negative for ANA. Compared with the ANA-negative group, the ANA-positive group showed significantly decreased incidence of cervical erythema (33.33% vs. 59.32%, P=0.010) and shawl sign (14.28% vs. 35.59%, P=0.017) . Twenty-eight (27.72%) patients with dermatomyositis were complicated by malignancies. Malignancies were found in 5 (11.9%) of ANA-positive patients, and in 23 (38.98%) of ANA-negative patients. Univariate analysis showed that ANA-negative patients with dermatomyositis had a higher risk of malignancies compared with ANA-positive patients with dermatomyositis, with an odds ratio of 7.52 (95% CI: 1.62-13.78, P=0.003) . In the multivariate regression model, the absence of ANA ( OR=4.34, 95% CI: 1.37-13.72, P=0.012) and cervical erythema ( OR=3.27, 95% CI: 1.20-8.91, P=0.020) were associated with high incidence of malignancies, while the absence of ANA was not significantly correlated with the occurrence of amyopathic dermatomyositis ( OR=0.99, 95% CI: 0.32-2.99, P=0.980) . Conclusions:ANA-negative adult dermatomyositis patients with cervical erythema had an increased risk of malignancies. Thus, close follow-up and regular tumor screening are necessary in these patients.

OBJECTIVE： To establish a method for simultaneous determination of nine components in Huoxiang zhengqi oral liquid， and to improve and perfect the quality standard of Huoxiang zhengqi oral liquid. METHODS： The contents of nine components in 10 batches of Huoxiang zhengqi oral liquid were determined by HPLC， such as licorice coumarin， isorlicin， liquiritinapioside， narirutin， liquiritin， saponins， hesperidin， magnolol and honokiol. The determination was performed on Kromasil Eternity XT-5-C18 column with mobile phase consisted of acetonitrile-0.05％ phosphoric acid solution （gradient elution） at the flow rate of 1 mL/min. The detection wavelength was set at 220 nm， and column temperature was 25 ℃. The sample size was 10 μL. RESULTS： The linear range of licorice coumarin， isorlicin， liquiritinapioside， narirutin， liquiritin， saponins， hesperidin， magnolol and honokiolin were 0.000 5-0.007 5， 0.000 8-0.025 0， 0.006 1-0.976 0， 0.001 6-0.250 0， 0.007 8-0.025 0， 0.000 4- 0.062 7， 0.008 6-0.276 0， 0.010 0-0.500 0， 0.010 0-0.500 0 mg/mL （r＝0.999 2-1.000 0）. The detection limits were 0.001 3， 0.000 1， 0.004 7， 0.005 0， 0.012 0， 0.001 3， 0.007 8， 0.007 7 0， 0.005 8 μg/mL， and the quantitative limits were 0.013 0， 0.000 8， 0.047 0， 0.050 0， 0.120 0， 0.013 0， 0.078 0， 0.070 0， 0.058 0 μg/mL， respectively； RSD of precision， stability and repeatability tests were less than 3.0％ （n＝6）. Average recovery rates were 98.67％， 101.85％， 98.97％， 103.05％， 100.00％， 97.78％， 97.91％， 100.13％， 101.95％； RSDs were 1.14％， 2.18％， 0.40％， 0.17％， 1.38％， 0.85％， 1.38％， 0.10％， 1.35％ （n＝6）. CONCLUSIONS： The established method is accurate and reliable， which can provide reference for the establishment of the overall quality control evaluation system and the improvement of quality standard for Huoxiang zhengqi oral liquid.