ObjectiveA mouse model of vaccinia virus Tiantan strain (VTT)-induced encephalopathy was developed using AG6 mice. MethodsVTT was amplified by infecting Vero cells at a multiplicity of infection (MOI) of 0.01, followed by concentration and titration. After 72 h of incubation, virus-containing cells were collected and subjected to concentration. The concentrated viral suspension was serially diluted (10-fold dilutions) and added to 6-well plates containing confluent Vero cell monolayers for plaque assay. The number of plaques formed in each well was counted, and the virus titer was calculated based on the dilution factor. Fourteen 5-6-week-old AG6 mice (half male and half female, housed separately by sex) were randomly divided into a control group (n=3, PBS), a low-dose group (n=6, 1×10⁵ PFU), and a high-dose group (n=5, 5×10⁵ PFU). The mice were anesthetized by isoflurane inhalation and then infected via intranasal instillation. The mental state of the mice in each group was observed daily, and the body weight and mortality were recorded. On day 13 post-infection, 2% Evans Blue (4 mL/kg body weight) was administered via tail vein injection to assess blood-brain barrier (BBB) disruption. Subsequently, brain tissue samples were collected for immunofluorescence analysis to evaluate the activation of astrocytes and microglia. ResultsThe titer of purified VTT was 1×10⁷ PFU/mL. Compared with the control group, mice in the low-dose group showed no significant change in body weight, and no lethality was observed. In contrast, mice in the high-dose group exhibited significant weight loss starting on day 5 post-infection (P<0.05), accompanied by lethality. On day 13 post-infection, no Evans Blue extravasation was detected in the brain tissues of the low-dose group, while the olfactory bulb region of the high-dose group displayed distinct blue staining, indicating disruption of the BBB. Immunofluorescence analysis revealed no significant proliferation of astrocytes and microglia in the olfactory bulb region of the low-dose group on day 13 post-infection. In contrast, marked activation of glial cells was observable in the high-dose group. ConclusionAn animal model of VTT-induced encephalopathy in AG6 mice is successfully established, characterized by BBB disruption and reactive gliosis specifically localized to the olfactory bulb region, manifested as astrocytic and microglial proliferation.

Objective To explore the impact of peak and duration of intrapartum fever on umbilical arterial blood gas analysis.Methods From January 2022 to December 2024,a total of 82 full-term singleton pregnant women after vaginal delivery in Hangzhou Women's Hospital with a system fever ≥38.0℃ were included as fever group.164 pregnant women with a body temperature＜38.0℃ during the same period were included as control group in a 1∶2 ratio.The umbilical arterial blood gas analysis was performed in all of them.General data,situation during childbirth and umbilical arterial blood gas analysis were compared between fever group and control group.The fever group was further divided into 3 subgroups,including 52 cases in subgroup A with temperature 38.0℃ to＜38.5℃,21 cases in subgroup B with temperature from 38.5℃ to＜39.0℃,9 cases in subgroup C with temperature ≥39.0℃.General data,situation during childbirth and umbilical arterial blood gas analysis of 3 subgroups were compared.Results There were no statistical significance in general data and situation during childbirth between fever group and control group(P＞0.05).Compared with control group,lower pH and base excess levels in umbilical artery blood as well as higher lactate levels in fever group with statistically significant differences(P＜0.05).There were no statistical significance in general data and situation during childbirth among subgroups(P＞0.05).pH,base excess and lactate levels in umbilical artery blood had no statistically significant differences among subgroups(P＞0.05).Logistic regression analysis showed that the fever duration was associated with umbilical artery acidosis(P=0.048,OR=1.005,95％CI:1.000-1.009).Conclusion Intrapartum fever leads to a decrease in pH and base excess in umbilical artery blood,as well as an increase in lactate levels.The peak of intrapartum fever leads no statistically significant differences in umbilical arterial blood gas analysis.For women with fever ≥38.0℃,controlling fever time may be helpful to reduce the incidence of acidosis and neonatal morbidity.

Objective To explore the effect of Xiongcan Yishen Formula on ferroptosis in testicular tissue of male rats with late-onset hypogonadism(LOH).Methods A total of 48 male Sprague-Dawley rats aged 6 months were randomly divided into model group,low-dose and high-dose Xiongcan Yishen Formula groups and propionate testosterone group,with 12 rats in each group.An additional 6 male Sprague-Dawley rats aged 8 weeks were set as the normal group.Except for the normal group,the rats were intraperitoneally injected with Cyclophosphamide at a dose of 20 mg/(kg·d)for 5 consecutive days to establish the LOH model,while the normal group received an equal volume of physiological saline for 5 days.The normal group and model group were given equal volumes of distilled water by gavage,while the low-dose and high-dose Xiongcan Yishen Formula groups were administered concentrated decoction at doses of 10.4 g and 41.6 g/kg/d respectively,and the propionate testosterone group received intramuscular injections of 5.21 mg/kg/d propionate testosterone,all for 28 consecutive days.ELISA was used to detect serum testosterone levels in rats,HE staining and transmission electron microscopy were used to observe the gross morphology of testicular tissue and the ultrastructure of Leydig cells,and RT-qPCR and Western blotting were used to detect the expression of ferroptosis-related genes and proteins in testicular tissue.Results The LOH model rats exhibited pathological changes such as atrophy of seminiferous tubules,structural disorder,and reduced spermatocytes in the lumen,as well as ultrastructural changes in Leydig cells including altered nuclear morphology,increased mitochondrial density,and reduced cristae.After intervention with Xiongcan Yishen Formula and propionate testosterone,the pathological changes in testis and the ultrastructure of Leydig cells were improved.Compared with the normal group,serum testosterone levels in the model group were significantly decreased(P＜0.05),the expression of ROS,ACSL4 mRNA and protein in testicular tissue was significantly increased,while the expression of FTH1,GPX4,and SLC7A11 mRNA and protein was significantly decreased(P＜0.05).Compared with the model group,ser-um testosterone levels in the low-dose and high-dose Xiongcan Yishen Formula groups and the propionate testosterone group were significantly increased(P＜0.05),and the expression of ROS,ACSL4 mRNA and protein was significantly decreased(P＜0.05);the high-dose Xiongcan Yishen Formula group showed significantly increased expression of FTH1,GPX4,and SLC7A11 mRNA and protein(P＜0.05).Conclusion Ferroptosis in testicular tissue is increased in LOH rats,and Xiongcan Yishen For-mula can elevate serum testosterone levels and improve pathological changes and ultrastructure in testicular tissue of LOH rats,possibly related to the inhibition of ferroptosis in testicular tissue of LOH rats.

Objective To construct mouse models of vaccinia virus infection using C57BL/6N and BALB/C mice infected with vaccinia virus VR-1354(WR(NIH TC-adapted)),and to compare the differences between the models in the two mouse strains.Methods Vaccinia virus VR-1354 was amplified,concentrated,and titrated,according to conventional method.The median lethal doses(LD50)of vaccinia virus V R-1354 for C57BL/6N and BALB/C mice were determined,respectively,and mice from the two strains were infected with the respective LD50.Lung tissues from control and experimental mice were removed for hematoxylin and eosin staining on days 3,7,and 14,and changes in the virus load in the lung tissues were measured at the same times.Results The titer of the amplified vaccinia virus VR-1354 was 3×108 plaque forming units(PFU)/mL.The LD50 values in C57BL/6N and BALB/C mice were 2.5×103and BALB/C 2.8×103PFU,respectively.After infecting mice from both strains with 2.5× 103 PFU of vaccinia virus VR 1354,lung tissue lesions worsened with increasing infection time,but the symptoms of the mice tended to recover after day 7.The virus loads in lung tissues began to increase on day 3 in both strains,peaked on day 7,and then gradually decreased.Conclusions C57BL/6N and BALB/C mouse models infected with vaccinia virus VR-1354 were established successfully.The virus can cause obvious lesions in mouse lung tissues,including inflammatory cell infiltration and alveolar cavity fluid exudation.The degree of disease onset and susceptibility differed between the two mouse strains,with C57BL/6N mice showing higher susceptibility to this virus than BALB/C mice.

Objective To study the predictive value of umbilical artery blood gas analysis combined with early postnatal myocardial enzyme spectrum for neonatal respiratory distress syndrome(RDS)and its severity in premature infants.Methods From January 2018 to December 2024,85 newborns with the gestation age of 30-36+6 weeks who were diagnosed as RDS after birth form Hangzhou Women's Hospital were selected as study group.85 newborns without diagnosed as RDS during the same period were included as control group in a 1:1 ratio.All of them born by cesarean section.The umbilical arterial blood gas analysis was performed and blood samples were collected for blood biochemical tests.The umbilical artery blood pH,base excess,lactate,and myocardial enzyme spectrum such as aspartate aminotransferase(AST),lactate dehydrogenase(LDH),creatine kinase isoenzymes(CK-MB)were compared between two groups.Binary Logistic regression was used to analyze the correlation between umbilical artery blood gas analysis and venous blood myocardial enzyme spectrum and RDS.Receiver operating characteristic curve of umbilical artery blood pH value and creatine kinase isoenzyme alone and their combined detection for predicting RDS was detected,and area under the curve(AUC),sensitivity and specificity of the single detection and combined detection were compared.The study group was further divided into subgroup A(mild)and subgroup B(severe)and the differences of the above indicators between two subgroups were compared.Results Compared with control group,tstudy group had lower pH and base excess levels in umbilical artery blood as well as AST,LDH and CK-MB were higher than those in control group with statistically significant differences(P＜0.05).pH and CK-MB were correlated with RDS(P＜0.05).The AUC of umbilical artery blood pH in predicting RDS was 0.728,the sensitivity was 0.588 and the specificity was 0.812.Combined with CK-MB,the AUC of predicting RDS was 0.813,the sensitivity was 0.729,and the specificity was 0.824.In the study group,subgroup A had higher pH and base excess levels in umbilical artery blood as well as AST,LDH and CK-MB were lower than those in subgroup B with statistically significant differences(P＜0.05).CK-MB was correlated with disease severity in newborns with RDS(P＜0.01).Conclusion Compared with simple application of pH,the combined use of CK-MB can more accurately predict RDS in premature infants,and also has a certain significance for the severity of RDS in premature infants.

Objective To construct mouse models of vaccinia virus infection using C57BL/6N and BALB/C mice infected with vaccinia virus VR-1354(WR(NIH TC-adapted)),and to compare the differences between the models in the two mouse strains.Methods Vaccinia virus VR-1354 was amplified,concentrated,and titrated,according to conventional method.The median lethal doses(LD50)of vaccinia virus V R-1354 for C57BL/6N and BALB/C mice were determined,respectively,and mice from the two strains were infected with the respective LD50.Lung tissues from control and experimental mice were removed for hematoxylin and eosin staining on days 3,7,and 14,and changes in the virus load in the lung tissues were measured at the same times.Results The titer of the amplified vaccinia virus VR-1354 was 3×108 plaque forming units(PFU)/mL.The LD50 values in C57BL/6N and BALB/C mice were 2.5×103and BALB/C 2.8×103PFU,respectively.After infecting mice from both strains with 2.5× 103 PFU of vaccinia virus VR 1354,lung tissue lesions worsened with increasing infection time,but the symptoms of the mice tended to recover after day 7.The virus loads in lung tissues began to increase on day 3 in both strains,peaked on day 7,and then gradually decreased.Conclusions C57BL/6N and BALB/C mouse models infected with vaccinia virus VR-1354 were established successfully.The virus can cause obvious lesions in mouse lung tissues,including inflammatory cell infiltration and alveolar cavity fluid exudation.The degree of disease onset and susceptibility differed between the two mouse strains,with C57BL/6N mice showing higher susceptibility to this virus than BALB/C mice.

Recent data suggest that vascular endothelial growth factor receptor inhibitor (VEGFRi) can enhance the anti-tumor activity of the anti-programmed cell death-1 (anti-PD-1) antibody in colorectal cancer (CRC) with microsatellite stability (MSS). However, the comparison between this combination and standard third-line VEGFRi treatment is not performed, and reliable biomarkers are still lacking. We retrospectively enrolled MSS CRC patients receiving anti-PD-1 antibody plus VEGFRi (combination group, n=54) or VEGFRi alone (VEGFRi group, n=32), and their efficacy and safety were evaluated. We additionally examined the immune characteristics of the MSS CRC tumor microenvironment (TME) through single-cell and spatial transcriptomic data, and an MSS CRC immune cell-related signature (MCICRS) that can be used to predict the clinical outcomes of MSS CRC patients receiving immunotherapy was developed and validated in our in-house cohort. Compared with VEGFRi alone, the combination of anti-PD-1 antibody and VEGFRi exhibited a prolonged survival benefit (median progression-free survival: 4.4 vs. 2.0 months, P=0.0024; median overall survival: 10.2 vs. 5.2 months, P=0.0038) and a similar adverse event incidence. Through single-cell and spatial transcriptomic analysis, we determined ten MSS CRC-enriched immune cell types and their spatial distribution, including naive CD4⁺ T, regulatory CD4⁺ T, CD4⁺ Th17, exhausted CD8⁺ T, cytotoxic CD8⁺ T, proliferated CD8⁺ T, natural killer (NK) cells, plasma, and classical and intermediate monocytes. Based on a systemic meta-analysis and ten machine learning algorithms, we obtained MCICRS, an independent risk factor for the prognosis of MSS CRC patients. Further analyses demonstrated that the low-MCICRS group presented a higher immune cell infiltration and immune-related pathway activation, and hence a significant relation with the superior efficacy of pan-cancer immunotherapy. More importantly, the predictive value of MCICRS in MSS CRC patients receiving immunotherapy was also validated with an in-house cohort. Anti-PD-1 antibody combined with VEGFRi presented an improved clinical benefit in MSS CRC with manageable toxicity. MCICRS could serve as a robust and promising tool to predict clinical outcomes for individual MSS CRC patients receiving immunotherapy.
Humans ; Colorectal Neoplasms/drug therapy* ; Male ; Female ; Immunotherapy ; Middle Aged ; Aged ; Tumor Microenvironment/immunology* ; Retrospective Studies ; Microsatellite Instability ; Transcriptome ; Single-Cell Analysis ; Programmed Cell Death 1 Receptor/immunology* ; Gene Expression Profiling ; Immune Checkpoint Inhibitors/therapeutic use* ; Adult ; Receptors, Vascular Endothelial Growth Factor/antagonists & inhibitors*

OBJECTIVE To analyze the quantity transfer rule in the processing of Astragalus membranaceus before and after moistening-soaking and steaming-soaking followed by cutting. METHODS Three batches of A. membranaceus decoction pieces processed through moistening-soaking and steaming-soaking followed by cutting were prepared. The HPLC overlapping fingerprints of A. membranaceus and its decoction pieces were established through the Similarity Evaluation System of Chromatographic Fingerprints of TCM （2012 edition）. Combined with the previous qualitative analysis results， the common peaks were identified， the changes of common peak area were analyzed， and the principal component analysis was carried out. The contents of calycosin-7-glucoside， astragaloside Ⅰ and astragaloside Ⅳ in A. membranaceus and its decoction pieces were determined by HPLC， and the content differences of each component in different samples were compared. RESULTS The results of fingerprint analysis showed that 17 common peaks were identified. After steaming-soaking and moistening-soaking of A. membranaceus， the proportion of common peak area in the decoction pieces changed compared with the original medicine （for example， in A. membranaceus steaming-soaking decoction pieces， the proportion of peak area of malonyl calycosin-7-glucoside and malonyl astragaloside Ⅰ decreased， while the proportion of peak area of calycosin-7-glucoside increased）. The results of principal component analysis showed that A. membranaceus， and its decoction pieces after moistening-soaking and steaming-soaking followed by cutting were all clustered into one category respectively. The results of content determination showed that， compared with A. membranaceus， the average content of calycosin-7-glucoside in A. membranaceus moistening-soaking decoction pieces was significantly reduced （P＜0.05）； the average contents of calycosin-7-glucoside and astragaloside Ⅳ in A. membranaceus steaming- soaking decoction pieces were significantly increased （P＜0.05）； there was no significant difference in the average content of astragaloside Ⅳ in A. membranaceus moistening-soaking decoction pieces and astragaloside Ⅰ in the two decoction pieces （P＞ 0.05）. CONCLUSIONS There are differences in the quantity transfer rules of A. membranaceus before and after moistening-soaking and steaming-soaking followed by cutting. Steaming-soaking followed by cutting may make the transformation of unstable components （such as malonyl calycosin-7-glucoside and malonyl astragaloside Ⅰ） more complete.

Objective To explore the environmental factors influencing the distribution of Pulsatilla chinensis(Bge.)Regel and to predict its potential habitat in China;To provide reference for the assessment of its wild resources and the introduction of its cultivation.Methods Totally 230 pieces of information on the distribution of sample sites were collected.Combined with data of 48 environmental factors,MaxEnt model and ArcGIS 10.8 software were used to select the dominant environmental factors that affect their distribution,and visual analysis of potential suitable areas was conducted.Results The prediction accuracy of the established model met the requirements(AUC＞0.9).The main environmental factors affecting the distribution of Pulsatilla chinensis(Bge.)Regel were the precipitation in the wettest season,the average temperature in November,the altitude,the standard deviation of the seasonal variation of temperature,the precipitation in May and the average temperature in August,with a cumulative contribution rate of 84.3％.The prediction results showed that it was mainly distributed in the North China and Northeast China,with a total suitability zone of about 2 382 461.60 km2,of which the low suitability zone was about 1 263 497.00 km2,the medium suitability zone was about 582 879.20 km2,and the high suitability zone was about 536 085.40 km2,accounting for approximately 53.03％,24.47％and 22.50％of the total suitability zone.Conclusion The results of the model are consistent with the literature records,which can provide theoretical reference for the sustainable utilization of Pulsatilla chinensis(Bge.)Regel.resources.

OBJECTIVE: To summarize the early clinical results and safety of Stand-alone OLIF application of lumbar lesions, and explored its surgical indications. METHODS: Total of 92 cases of lumbar spine lesions treated with Stand-alone OLIF at two medical centers from October 2014 to December 2018 were retrospectively analyzed, including 30 males and 62 females with an average age of (61.20±12.94) years old ranged from 32 to 83 years old. There were 20 cases of lumbar spinal stenosis, 15 cases of lumbar disc degeneration, 11 cases of lumbar degenerative spondylolisthesis, 6 cases of discogenic low back pain, 7 cases of giant lumbar disc herniation, 13 cases of primary lumbar discitis, 6 cases of adjacent vertebral disease after lumbar internal fixation surgery, and 14 cases of degenerative lumbar scoliosis. Pre-operative dual energy X-ray bone density examination 31 cases' T-values ranged from -1 to -2.4, 8 cases' T-values ranged from -2.5 to -3.5, and the rest had normal bone density. The number of fusion segments: 68 cases of single segment, 9 cases of two segment, 12 cases of three segment , and 3 cases of four segment. Fusion site:L_1,2 1 case, L_2,3 4 cases, L_3,4 10 cases, L_4,5 53 cases, L_2,3-L_3,4 3 cases, L_3,4-L_4,5 6 cases, L_1,2L_2,3L_3,4 1 case, L_1,2L_3,4L_4,5 1 case, L_2,3L_3,4L_4,5 10 cases, L_1,2L_2,3L_3,4L_4,5 3 cases. The clinical results and imaging results of this group of cases were observed, as well as the complications. RESULTS: The surgical time ranged from 40 to 140 minutes with an average of (60.92±27.40) minutes. The intraoperative bleeding volume was 20 to 720 ml with an average of (68.22±141.60) ml. The patients had a follow-up period of 6 to 84 months with an average of (38.50±12.75) months. The height of the intervertebral space recovered from (9.23±1.94) mm in preoperative to (12.68±2.01) mm in postoperative, and (9.11±1.72) mm at the last follow-up, there was a statistically significant difference(F=6.641, P=0.008);there was also a statistically significant difference between the postoperative and preoperative height of the intervertebral space(t=9.27, P<0.000 1);and there was also a statistically significant difference (t=10.06, P<0.000 1) between the last follow-up and postoperative height of the intervertebral space. At the last follow-up, cage subsidence grading was as follows:level 0 in 69 cases (76 segments), levelⅠin 17 cases (43 segments), level Ⅱin 5 cases (14 segments), and level Ⅲ in 1 case (1 segment);according to the number of segments, normal subsidence accounts for 56.72%, abnormal subsidence accounts for 43.28%. Bone mineral desity of normal subsidence groups was -0.50±0.07 whinch was better than that the abnormal subsidence groups -2.10±0.43, and the difference was statistically significant(χ²=2.275, P=0.014). As well as there was a statistically significant difference in the patient's VAS of backache from (6.28±2.11) in preoperative to (1.48±0.59) in last follow-up(t=8.56, P<0.05). The ODI recovered from (36.30±7.52)% before surgery to (10.20±2.50)% at the last follow-up, with a statistically significant difference (t=7.79, P<0.000 1). Complications involved 4 cases of intraoperative vascular injury, 21 cases of endplate injury, and 4 cases of combined vertebral fractures. The incision skin has no necrosis or infection. There were 4 cases of left sympathetic chain injury, 4 cases of transient left hip flexion weakness, 2 cases of left thigh anterolateral numbness with quadriceps femoris weakness, and 1 case of incomplete intestinal obstruction;8 cases were treated with posterior pedicle screw fixation due to fusion cage settlement accompanied by stubborn lower back pain, and 6 cases were treated with fusion cage settlement and lateral displacement. According to the actual number of cases, there were 38 complications, with an incidence rate of 41.3%. CONCLUSION The application of Stand alone OLIF in lumbar spine disease fusion has achieved good early results, with obvious clinical advantages, but also there are high probability of complications. It is recommended to choose carefully. It is necessary to continuously summarize and gradually clarify and complete the surgical indications and specific case selection criteria.
Humans ; Male ; Female ; Middle Aged ; Spinal Fusion/methods* ; Lumbar Vertebrae/injuries* ; Aged ; Adult ; Retrospective Studies ; Aged, 80 and over