AIM To establish LC-MS and GC-MS method and analyze the chemical constituents of Dendrobium huoshanense C.Z.Tang & S.J.Cheng flowers.METHODS LC-MS was performed on a Zorbax Eclipse C18 column(2.1 mm×100 mm,1.8 μm),with the mobile phase comprising of water(containing 0.1％formic acid)-acetonitrile flowing at 0.3 mL/min,and electrospray ionization was operated in both positive and negative ion modes.The GC-MS employed headspace solid-phase microextraction for sample preparation,and the analysis was performed on an HP-5MS column(30 m×0.25 mm,0.25 μm),with the following temperature program:initial temperature 50 ℃(held for 2 min),increased at 5 ℃/min to 180 ℃(held for 5 min),then raised at 10 ℃/min to 250 ℃(held for 5 min),and electron impact ion source was employed.RESULTS A total of 62 compounds were identified by LC-MS,including 35 flavonoids,4 coumarins,6 alkaloids,6 terpenoids,3 amino acids,2 polyphenols,2 ketones and 4 others.A total of 101 volatile components were identified by GC-MS,including ketones,aldehydes,alcohols,esters,ethers,and acid.CONCLUSION This method can comprehensively analyze the chemical constituents of D.huoshanense flowers,and provide a scientific basis for elucidating its pharmacodynamic material basis.

Objective:To analyze effects of T11TS in the treatment of Cryptococcus neoformans fungal infection and the regula-tion of T-cell calcineurin(CaN)-activated T lymphoid nuclear factor(NFAT)pathway.Methods:SD rats infected with Cryptococcus neoformans were treated with immunoenhancer T11TS.Flow cytometry,Western blot and nuclear translocation were used to detect the changes of T cell function and related signal pathway.Results:The number of Cryptococcus neoformans in lung and spleen of CT3 group was significantly less than Cryptococcus neoformans infection group,CT1 group and CT2 group(P<0.05).The results of flow cy-tometry showed that the expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 in Cryptococcus neoformans infection group were significantly lower than control group(P<0.05).The expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 of CT1,CT2 and CT3 groups were significantly higher than Cryptococcus neoformans infection group(P<0.05).The results of Western blot were similar to those of flow cytometry.Conclusion:T11TS in the treatment of Cryptococcus neoformans infection can enhance the immunity of the body by up-regulating the expression of related signal factors in the CaN-NFAT pathway of T lymphocytes,increasing the expression of NFAT and IL-2,stimulating the activation and proliferation of T lymphocytes.

Objective To establish a stable,reliable,and clinically relevant porcine model of endotoxin-induced acute respiratory distress syndrome(ARDS).Methods Ten 8-month-old male Bama minipigs were deeply sedated,followed by invasive mechanical ventilation and electrocardiographic monitoring.Lipopolysaccharide(LPS)was intravenously pumped at 600 μg/(kg·h)for 3 hours,then maintained at 15 μg/(kg·h)thereafter.Dynamic monitoring was performed at five time points after LPS injection(LPS 0,1,3,5,and 8 h),including arterial blood gas analysis and chest computed tomography(CT)scans.Pathological examination of lung tissues obtained via bronchoscopic biopsy(HE staining and transmission electron microscopy)was conducted.These indicators were comprehensively used to evaluate the success of the animal model.Results At 5 hours after LPS administration,8 minipigs developed symptoms such as skin cyanosis,elevated body temperature,and respiratory distress.The oxygenation index decreased to<300 mmHg.Chest CT scans showed diffuse pulmonary infiltrates.Histopathology revealed alveolar edema and hyaline membrane formation.Transmission electron microscopy demonstrated disruption of pulmonary blood-air barrier,depletion of lamellar bodies in type Ⅱ pneumocytes,inflammatory cell infiltration,and exudation of plasma proteins and fibrin.Compared with LPS 0 h,at LPS 8 h,the oxygenation index and arterial blood pH were significantly decreased(P<0.001),while blood lactic acid and serum potassium were significantly increased(P<0.05);serum calcium and base excess were significantly decreased(P<0.05),and the lung injury score based on HE-stained lung sections was significantly increased(P<0.01).Conclusion The porcine ARDS model established by continuous LPS injection can dynamically simulate the pathophysiological characteristics and typical pathological manifestations of clinical septic ARDS,making it an effective tool to study the pathogenesis,prevention,and treatment strategies of septic ARDS.

Objective To discuss the application of bidirectional dual loop technique in retrieving Aegisy inferior vena cava filters.Methods The clinical data of 12 patients with deep vein thrombosis in the lower limbs complicated by pulmonary embolism,who received mechanical thrombectomy and Aegisy inferior vena cava(IVC)filter implantation at the Zhoukou Municipal Central Hospital of China from January 2016 to November 2023,were retrospectively analyzed.The retrieval of Aegisy IVC filters beyond the time window was tried by standard methods,guide wire stripping,balloon dilatation,unidirectional single loop technique,unidirectional dual loop technique,etc.However,because of the difficulty in retrieval of filter,bidirectional dual loop technique had to be used.The indwelling time of the IVC filters was 30-365 days,with an average time of 105 days.Results The IVC filter was completely removed in 11 patients.Angiography showed that IVC was unobstructed,no contrast extravasation was observed,and the average operation time was 126 minutes.The retrieval of filter failed in one patient,and the patient had to receive anticoagulation therapy lifelong.No surgery-related complications occurred.Conclusion For retrieving Aegisy IVC filters beyond the time window,bidirectional dual loop technique is clinically safe and effective,and this technology is worthy of clinical promotion and application.

Objective To investigate the density,species composition,and seasonal prevalence of domestic rodents in different habitats within Lanzhou garrisons,providing basic information for rodent prevention and control.Methods A total of 12 monitoring sites were sampled across urban,suburban,and rural residential areas from 2014 to 2022.Rodent density was monitored using the night-trapping method in the middle of odd-numbered months.Results From 2014 to 2022,346 domestic rodents were captured using 122 035 effective traps,with an average domestic rodent density of 0.28％.The highest domestic rodent density was 0.63％in 2016,and the lowest was 0.07％in 2020,showing significant differences across years,with an overall trend of initially decreasing and then increasing(χ2=136.555,P<0.001).The dominant species was Rattus norvegicus,accounting for 83.24％of the total rodents captured.Rattus norvegicus accounted for a relatively high proportion across different years,with a statistically significant difference in species composition(χ2=20.931,P<0.05).Rodent densities and species composition also varied significantly among the monitored habitats(P<0.001),with the highest densities observed in rural residential areas and the lowest in urban areas.Seasonal variation in rodent densities showed a bimodal pattern,with smaller peaks in January or March and a larger peak in July.Conclusions Domestic rodent density in Lanzhou garrisons has shown an upward trend in the past few years.Rodent control measures should focus on barracks in rural residential areas,with targeted interventions to reduce the risk of rodent-borne diseases.

Objective:To investigate the clinical manifestation, therapy, and prognosis of Susac syndrome and enhance the understanding of this disease.Methods:A case summary was made.The clinical data of two siblings with Susac syndrome treated at Children′s Medical Center, Peking University First Hospital in January 2024 were summarized.Reported cases of pediatric Susac syndrome were reviewed.Results:The onset of the disease in the two siblings was at the age of 3.00 and 6.75 years, with recurrent headaches, tinnitus, hearing loss and encephalopathy symptoms.Cranial magnetic resonance imaging showed multiple cerebral microbleeding and microinfarction lesions, " snowball like" in the corpus callosum and diffuse white matter edema in the brain.Audiometry revealed sensorineural hearing loss.In one case, ophthalmic fluorescein angiography revealed ischemic changes due to branch retinal artery occlusions.No pathogenic variants were detected in gene testing.This child was diagnosed with Susac syndrome, and the symptoms were improved after treatment with Corticosteroids and Rituximab.No relapse was observed during the 9-month follow-up.A total of 20 pediatric cases of Susac syndrome were retrieved, including 18 reported previously and 2 cases from this study.There were 2 boys and 18 girls, with the age of onset ranging from 2.5 to 17.0 years.The common initial symptoms included headache (19 cases), vertigo and tinnitus or hearing loss (9 cases), and vision impairment or visual field defect (4 cases). The symptoms were improved after immunotherapy.Conclusions:With a low incidence, Susac syndrome is rare in children and difficult to diagnose.There may be a genetic predisposition in such disease.Early diagnosis and immunotherapy can low the relapse and improve the prognosis.

Objective:To analyze effects of T11TS in the treatment of Cryptococcus neoformans fungal infection and the regula-tion of T-cell calcineurin(CaN)-activated T lymphoid nuclear factor(NFAT)pathway.Methods:SD rats infected with Cryptococcus neoformans were treated with immunoenhancer T11TS.Flow cytometry,Western blot and nuclear translocation were used to detect the changes of T cell function and related signal pathway.Results:The number of Cryptococcus neoformans in lung and spleen of CT3 group was significantly less than Cryptococcus neoformans infection group,CT1 group and CT2 group(P<0.05).The results of flow cy-tometry showed that the expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 in Cryptococcus neoformans infection group were significantly lower than control group(P<0.05).The expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 of CT1,CT2 and CT3 groups were significantly higher than Cryptococcus neoformans infection group(P<0.05).The results of Western blot were similar to those of flow cytometry.Conclusion:T11TS in the treatment of Cryptococcus neoformans infection can enhance the immunity of the body by up-regulating the expression of related signal factors in the CaN-NFAT pathway of T lymphocytes,increasing the expression of NFAT and IL-2,stimulating the activation and proliferation of T lymphocytes.

Objective To compare the capture performance differences between TruSeq? Exome and NimbleGen SeqCap EZ Human Exome kits in children with cerebral palsy(CP),and to provide a technical selection basis for clinical genetic research and diagnosis.Methods Peripheral blood samples from 48 sporadic CP patients were included.Exome libraries were constructed using TruSeq(DNA probes)and NimbleGen(RNA probes),followed by sequencing on the Illumina HiSeq 2000 platform.Bioinformatics analysis was applied to evaluate mapping rate,target region coverage,variant concordance,and clinical relevance based on a CP-related gene set(2 293 genes).The statistical analysis was performed using a paired t-test with a significance threshold of α=0.05.Results The results showed no significant differences between NimbleGen and TruSeq exome capture kits in basic data quality(alignment rate,insert size)and GC content.However,they exhibited complementary characteristics in key performance metrics:NimbleGen demonstrated superior performance in specific depth coverage(1×coverage rate,P=1.84×10-5;20×coverage rate,P=1.49×10-20).TruSeq,on the other hand,showed higher sensitivity in Indel detection(TruSeq vs.NimbleGen:11 371±1 689 vs.11 274±1 670,P=3.24×10-7)and rare variant capture(TruSeq vs.NimbleGen:3 164±766 vs.3 072±774,P=1.20×10-4),successfully identifying all 11 pathogenic variants(including 2 missed by NimbleGen).Conclusion TruSeq,with its superior variant detection rate,is more suitable for clinical diagnostic applications,while NimbleGen's coverage stability may be advantageous for research-oriented projects.

Objective:To study the growth inhibition of Thunberg Fritillary extract on non-small cell lung cancer.Methods:The Thunberg Fritillary extract was prepared and characterized by UPLC-QE/MS.Replicated Lewis lung carcinoma ectopic tumor-bear-ing mouse model,yeast injection induced acute inflammation,compared the effect of Thunberg Fritillary extract combination with acute inflammation on the growth,tumor volume and tumor suppression rate of Lewis lung carcinoma mice,and determine the content of inflammatory factors by the flow CBA method(IL-6,IL-1β,IL-1α,IL-10,IL-27,IL-17A,IL-12p70,IL-23,TNF-α,IFN-γ,IFN-β,GM-CSF,MCP-1).Results:The inhibition of Lewis lung carcinoma mice was similar to that of cisplatin alone,and the tumor suppression rate was 35%;the tumor suppression rate of Thunberg Fritillary extract combined with acute inflammatory stimulation of yeast was 62%,1.8 times that of cisplatin alone.The decrease in the expressions of cytokines IL-23,MCP-1 after acute inflammatory stimulation in yeast was associated with tumor suppression;while the increased expressions of IL-6,IL-1β,IL-1α,IL-10,IL-27,IL-17A,IL-12p70,TNF-α,IFN-γ,IFN-β and GM-CSF cytokines were associated with tumor suppression.Conclusion:The Thun-berg Fritillary extract combination with acute inflammation can play a positive role against non-small cell lung cancer,which will pro-vide new research ideas and methods for the prevention and treatment of non-small cell lung cancer.

Objective To compare the capture performance differences between TruSeq? Exome and NimbleGen SeqCap EZ Human Exome kits in children with cerebral palsy(CP),and to provide a technical selection basis for clinical genetic research and diagnosis.Methods Peripheral blood samples from 48 sporadic CP patients were included.Exome libraries were constructed using TruSeq(DNA probes)and NimbleGen(RNA probes),followed by sequencing on the Illumina HiSeq 2000 platform.Bioinformatics analysis was applied to evaluate mapping rate,target region coverage,variant concordance,and clinical relevance based on a CP-related gene set(2 293 genes).The statistical analysis was performed using a paired t-test with a significance threshold of α=0.05.Results The results showed no significant differences between NimbleGen and TruSeq exome capture kits in basic data quality(alignment rate,insert size)and GC content.However,they exhibited complementary characteristics in key performance metrics:NimbleGen demonstrated superior performance in specific depth coverage(1×coverage rate,P=1.84×10-5;20×coverage rate,P=1.49×10-20).TruSeq,on the other hand,showed higher sensitivity in Indel detection(TruSeq vs.NimbleGen:11 371±1 689 vs.11 274±1 670,P=3.24×10-7)and rare variant capture(TruSeq vs.NimbleGen:3 164±766 vs.3 072±774,P=1.20×10-4),successfully identifying all 11 pathogenic variants(including 2 missed by NimbleGen).Conclusion TruSeq,with its superior variant detection rate,is more suitable for clinical diagnostic applications,while NimbleGen's coverage stability may be advantageous for research-oriented projects.