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 explore the improvement effect and mechanism of salidroside on radiation-induced parotid gland injury in rats. METHODS Rats were randomly assigned into normal group， radiation group， salidroside low-dose （salidroside-L， 50 mg/kg） group， salidroside high-dose （salidroside-H， 100 mg/kg） group， and salidroside-H+inhibitor （100 mg/kg salidroside+0.1 µmol/kg H-89） group， with 10 rats in each group. Except for the normal group， rats in the other groups were subjected to radiation exposure to establish a model of radiation-induced parotid gland injury. Rats in each group were intraperitoneally injected with the corresponding drug or normal saline， once a day， for 40 consecutive days. After the last administration， the levels of reactive oxygen species （ROS）， cyclic adenosine monophosphate （cAMP）， superoxide dismutase （SOD）， and amylase in serum were detected； the pathological changes of parotid gland tissue were observed； the apoptosis rate of parotid gland tissue cells， the expression levels of B-cell lymphoma-2 （Bcl-2） and its associated X protein （Bax）， mRNA expression levels of interleukin-6 （IL- 6） and tumor necrosis factor-α （TNF-α）， the protein expression levels of type Ⅲ collagen （Col Ⅲ）， vasoactive intestinal peptide （VIP）， and the phosphorylation level of protein kinase A （PKA） in parotid gland tissue were determined. RESULTS Compared with normal group， the levels of ROS， amylase， apoptosis rate， Bax expression level， mRNA expression levels of IL-6 and TNF- α， and protein expression level of Col Ⅲ in the radiation group were significantly increased， while the levels of cAMP， SOD， Bcl-2 expression level， VIP protein expression level and PKA phosphorylation level were significantly decreased （P＜0.05）. Compared with radiation group， the above indicators in the salidroside-L group and salidroside-H group were significantly improved （P＜0.05）， and the improvement in the salidroside-H group was more significant （P＜0.05）； inhibitor H-89 significantly reversed the changes in the above indicators of the salidroside-H group （P＜0.05）. CONCLUSIONS Salidroside can effectively alleviate radiation-induced parotid gland injury in rats， and its mechanism may be related to the activation of the VIP-cAMP pathway.

Objective To systematically evaluate the incidence rate of low-level viremia(LLV)in chronic hepatitis B(CHB)patients and related influencing factors,and to provide evidence-based medicine evidence for effective intervention and prevention of LLV in clinical practice.Methods This study was conducted according to the PRISMA guideline,with a PROSPERO registration number of CRD42023455304.CNKI,Wanfang Data,VIP,SinoMed,PubMed,Embase,Web of Science,and the Cochrane library were searched for observational studies on LLV and related influencing factors in CHB patients published up to July 21,2023.Stata 16.0 software was used to perform the meta-analysis.Results A total of 12 articles were included,with a total sample size of 3408 cases,among whom there were 1181 patients with LLV.The meta-analysis showed that the incidence rate of LLV was 32.8%(95%confidence interval[CI]:27.6%—38.3%)in treatment-experienced CHB patients.High HBsAg quantification(odds ratio[OR]=2.107,95%CI:1.782—2.491,P<0.001),positive HBeAg(OR=3.258,95%CI:2.629—4.038,P<0.001),high HBV DNA level at baseline(OR=1.286,95%CI:1.157—1.430,P<0.001),and history of entecavir treatment(OR=3.089,95%CI:1.880—5.074,P<0.001)were risk factors for LLV;duration of antiviral therapy≥3 years(OR=0.175,95%CI:0.093—0.331,P<0.001)and high alanine aminotransferase level at baseline(OR=0.985,95%CI:0.978—0.992,P<0.001)were protective factors against LLV.The sensitivity analysis showed no significant change in effective value,suggesting that the results of the meta-analysis were relatively stable.The funnel plot of the studies included was basically symmetrical,and the results of the Egger's test and the Begg's test suggested that there was no obvious publication bias in the articles included.Conclusion Clinicians should guide decision making based on the influencing factors for LLV and related clinical evidence,so as to reduce long-term clinical risks and avoid adverse outcomes.

In order to establish a rapid,specific and sensitive detection method for Streptococcus equi subspecies zooepidemicus(SEZ)and to understand the infection status of SEZ in horses ente-ring China,specific primers were designed and synthesized based on the conserved gene comB of standard strain SEZ(ATCC 43079)in this work.Then,the pMD19-T-comB recombinant plasmid was constructed and used as a standard positive template.After that,the fluorescence-based quantitative PCR(qPCR)detection method based on SYBR Green Ⅰ dye was established.Totally,477 equine entry serum samples from 6 countries,including Netherlands,Belgium,Japan,Germa-ny,Argentina and New Zealand,during 2018 to 2023,were randomly selected and detected for SEZ by the qPCR method.Results showed that the established qPCR method had specific amplification for only SEZ,which illustrated a good specificity.Sensitivity test of the method showed that the limited detection amount was 4.58 X101 copies/μL.And the repeatability test showed that the coef-ficient of variation of intra-batch repeatability was less than 0.5％,while the inter-batch repeat-ability was less than 3.0％,which indicated good repeatability and high stability.Retrospective a-nalysis showed that totally 11 of 477 positive samples were detected,with a relatively low positive rate of 2.31％(11/477).Among them,all the 40 samples from Netherlands in 2018 were negative(0/40).In the samples of 2019,one positive was detected from Belgium(1/20),while all other 36 samples which form Japan and Germany were negative.In the samples of 2021,three samples(3/34)from Japan and one sample(1/20)from Argentina were positive,and all the other 40 samples from the Netherlands were negative.In the samples of 2022,76 samples from Netherlands were all negative.While in the 2023,5(5/126)of 126 samples from Netherlands and one(1/88)of 88 from New Zealand were found positive with SEZ.To summarize,The SYBR Green Ⅰ qPCR method for the diagnosis of SEZ was successfully established,and it could provide necessary technical support for the rapid quarantine of China's entry-exit and port departments,as well as the epidemiological investigation of the disease.

In order to understand the prevalence of Akabane disease(AKAD)in Guizhou Province and the molecular characteristics of the isolates,the whole-genome sequence of a strain of Akabane virus(AKAV)from a bovine AKAD-positive sample was determined and analyzed.The genotype and genetic variation of the strain were also explored.Based on the conserved S sequence,a fluores-cence quantitative PCR(qPCR)detection method was established and applied for the investigation of AKAV infection status in four large-scale beef cattle farms of Guizhou.Results showed that the S,M and L fragments of the bovine strain were highly homologous to the Tianjin strain(TJ2016/China/2016)and the Australian strain(JaLAB39/Australia/1959),where they were in the same evolutionary branch and belonged to genotype Ⅱ.Sensitivity assay found that the lowest detection limit was 2.5 X 101 copies/μL.Specificity assay showed the established method detected only AKAV with no amplification on bovine bluetongue virus(BLUV),Pasteurella multocida(PM),bovine infectious rhinotracheitis virus(IBRV)and bovine Mycoplasma bovis.The variation coefficients of inter-and intra batches in the repeatability test were both lower than 2.26％.These findings illus-trated that the established qPCR method had high sensitivity,good specificity and repeatability.A total of 298 serum samples from 4 large-scale beef cattle farms in Qianxi City and Huangping County of Guizhou Province were collected and tested for AKAV by the method.Out of 298 sam-ples,25 positive samples(25/298)were detected as positive with a positive rate of 8.39％.In sum-mary,this work provided the reference data for a deep understanding of the molecular prevalence of AKAV in Guizhou Province and laid foundation for the prevention and control of AKAD.

Chest trauma is one of the most common injuries. Venous thromboembolism (VTE) as a common complication of chest trauma seriously affects the quality of patients′ life and even leads to death. Although there are some consensus and guidelines on the prevention and treatment of VTE at home and abroad, the current literatures lack specificity considering the diagnosis, treatment and prevention of VTE in patients with chest trauma have their own characteristics, especially for those with blunt trauma. Accordingly, China Chest Injury Research Society and editorial board of Chinese Journal of Traumatology organized relevant domestic experts to jointly formulate the Chinese expert consensus on the diagnosis, treatment and prevention of chest trauma venous thromboembolism associated with chest trauma (2022 version). This consensus provides expert recommendations of different levels as academic guidance in terms of the characteristics, clinical manifestations, risk assessment, diagnosis, treatment, and prevention of chest trauma-related VTE, so as to offer a reference for clinical application.

Objective: To detect the clinicopathological features, immunophenotype, diagnosis, and differential diagnosis of composite pheochromocytoma(CP). Methods: Five cases of CP were collected at Zhejiang Provincial People′s Hospital from January 2011 to January 2019. The clinical, radiological, histologic, immunohistochemical and outcome data were analyzed; the diagnosis and differential diagnosis were discussed. Results: The patients′ age range was 52-68 years (mean 59 years, median 54 years), There were 4 males and 1 female, and the male to female ratio was 4∶1. Tumor size was 3-4 cm (mean 3.6 cm, median 3.5 cm). The most common clinical manifestation was adrenal mass. Histologically, the classical feature was two distinct morphologic components, one with tumor cells arranged in irregular nests, and with fine granular and basophilic oramphophilic cytoplasm; the other was composed of scattered ganglion cells in the background of Schwann cells organized in interwoven bundles. The components of pheochromocytoma expressed PHOX2B(5/5), synaptophysin (5/5), CgA (5/5), the sustentacular cells expressed S-100 protein; the components of ganglioneuroma expressed S-100 protein (5/5), NF (5/5), the ganglion cells were weakly positive for PHOX2B, synaptophysin and CgA. All the cases were surgically resected and all patients were free of recurrence at follow-up. Conclusions CP is rare adrenal tumor, and it has typical histologic features but no specific clinical manifestations. Attention should be paid to its characteristic histomorphology with the use of PHOX2B, CgA, synaptophysin and S-100 protein immunohistochemistry that is helpful for its diagnosis.

Objective To explore the value of CT images in distinguishing perihematomal edema in basal ganglia intracerebral hemorrhage with normal brain tissue,and its significance in assessing patients' conditions and prognoses.Methods CT images and clinical data of 120 patients with basal ganglia intracerebral hemorrhage admitted to our hospital from January 2017 to September 2018 were collected,and these 120 patients were randomly assigned to group of training data set (n=90) and group of test data set (n=30) at a ratio of 3∶l.The texture analysis software Mazda was used to preprocess the CT images and manually sketch the regions of interest (ROIs) to extract the texture parameters in patients from the group of training data set;Mazda software provides texture feature selection methods including mutual information (MI),Fisher coefficients (Fisher),classification error probability combined with average correlation coefficients (POE+ACC),and texture feature analysis including raw data analysis (RDA),principal component analysis (PCA),linear classification analysis (LDA) and nonlinear classification analysis (NDA);texture feature selection methods and texture feature analysis were grouped by pairs to establish different image omics labels;the error rate was used to evaluate the performance of different labels.Random forest model,support vector machine model and neural network model were built for texture parameters in patients from the group of test data set,and texture parameters extracted from patients from group of training data set were imported into these models;receiver operating characteristics curve was used to assess the performance of models.According to the maximum diameter of the hematomas,Glasgow coma scale (GCS) scores at admission,median of National Institute of Health Stroke Scale (NIHSS) scores 3 months after follow up,all patients were divided into two groups;Mazda software was used repeatedly for dimension reduction and establishment of different images omics labels;the sum of error rates from the two groups was taken as total error rate to evaluate the significance of different labels in predicting patients' conditions and prognoses.Results A total of 295 texture parameters were extracted from the ROIs of the best CT images of 90 patients from group of training data set,and 10 characteristic texture parameters were obtained by each of the three dimensionality reduction methods.Among all texture post-processing methods,the lowest error rate was 2.22％ for POE+ACC/NDA;AUCs were 0.87 (95％ CI:0.76-0.97),0.81 (95％ CI:0.72-0.93) and 0.76 (95％CI:0.67-0.89) for random forest model,support vector machine model and neural network model in the test dataset,respectively,which indicated that random forest model had the best forecast performance.The imaging omics labels established based on POE+ACC/NDA had the lowest total error rate for analysis of maximum diameter ofhematoma and GCS scores at admission (26.66％,23.33％);the imaging omics labels established based on Fishers coefficient method and NDA had the lowest total error rate(33.33％) for analysis of NIHSS scores at 3 months of follow up.Conclusion Radiomicmethod with proper model is of certain value in distinguishing erihematomal edema in basal ganglia intracerebral hemorrhage with normal brain tissue,and also has certain significance in evaluating the patienfs conditions and prognoses.

Objective To investigate the clinical effects and safety differences of endoscopic hormone injection by tympanic membrane puncture and eustachian tube in the treatment of patients with chronic secretory otitis media.Methods From January 2015 to September 2016,90 patients with chronic secretory otitis media in Zhoushan Hospital were chosen.They were randomly divided into two groups according to the digital table.The tympanic membrane puncture group (65 patients) was treated with hormone injection by tympanic membrane puncture and the eustachian tube group(65 patients) was treated with endoscopic hormone injection by eustachian tubeThe total clinical effective rate,the bone conduction threshold value in 1 kHz,2kHz,4kHz and 8kHz,the ETDQ-7 score and the levels of laboratory index before and after treatment,the secondary tympanic effusion rate and the recurrence rate with follow-up of both groups were compared.Results The total clinical effective rate of the tympanic membrane puncture group and eustachian tube group were 77.78％ and 93.33％,respectively.The total clinical effective rate of the eustachian tube group were significant higher than that of the tympanic membrane puncture group (x2 =9.84,P ＜ 0.05).The bone conduction threshold value in l kHz,2kHz,4kHz and 8kHz of the tympanic membrane puncture group after treatment were (10.42 ± 1.60) kHz,(12.86 ± 2.50) kHz,(16.09 ± 2.81) kHz,(15.26 ± 2.68) kHz,respectively.The bone conduction threshold value in 1kHz,2kHz,4kHz and 8kHz of the eustachian tube group after treatment were (9.75 ± 1.36) kHz,(11.13 ± 2.02) kHz,(14.82 ± 2.44) kHz,(13.78 ± 2.11) kHz,respectively.The bone conduction threshold value in 1kHz,2kHz,4kHz and 8kHz of the eustachian tube group after treatment were significant lower than those of the tympanic membrane puncture group and those before treatment(t =3.10,3.56,3.29,3.96,2.87,3.15,2.91,3.28,2.67,2.81,2.78,2.50,all P ＜ 0.05).The ETDQ-7 scores of the eustachian tube group after treatment were significant lower than those of the tympanic membrane puncture group and those before treatment(t =3.31,3.87,2.89,all P ＜ 0.05).The levels of laboratory index of the eustachian tube group after treatment were significant higher than those of the tympanic membrane puncture group and those before treatment (t =3.36,3.77,3.71,4.02,2.83,3.06,all P ＜ 0.05).The secondary tympanic effusion rate and the recurrence rate with follow-up of the eustachian tube group were significant lower than those of the tympanic membrane puncture group(x2 =9.87,12.33,10.67,all P ＜ 0.05).Conclusion Compared with endoscopic hormone injection by tympanic membrane puncture,endoscopic hormone injection by eustachian tube in the treatment of patients with chronic secretory otitis media possess the advantages including relieve the clinical symptoms,improve the bone conduction threshold and eustachian tube function,regulate the laboratory indexes and reduce the risk of recurrence and effusion.

Objective To evaluate the diagnostic value of ferritin,procalcitonin (PCT) and C-reactive protein (CRP) on infectious diseases.Methods A total of 190 patients with infectious diseases in our hospital from 2012 to 2017 were enrolled and analysed retrospectively.Among them,bacterial infection was found in 130 cases (bacterial infection group),and viral infection in 60 cases (viral infection group).Serum levels of erritin,CRP,PCT in 190 patients were deteceted for further statistical analysis.Results Among the 190 cases of infectious fever,serum levels of erritin,CRP,PCT in bacterial infection group were higher than that in the viral infection group,with significant difference.The receiver operating curve (ROC) curve analysis showed that area under curve (AUC) of CRP,PCT,ferritin were 0.797,0.803,0.883.Conclusions The present study identified ferritin,PCT and CRP that should be considered in the differential diagnosis of infectious diseases,providing useful information for future diagnosis.