The study was aimed at identifying the diversity of tick species in selected areas of Qinghai,to analyze the genetic differentiation characteristics of tick-borne spotted fever group rickettsiae(SFGR),and to provide the theoretical basis for SFGR prevention and control in the region.The 16S rRNA gene was used for molecular biological identification of 446 collected tick samples,and the infection characteristics of SFGR in tick samples were determined according to the SFGR outer membrane protein A(ompA)gene.Haplotype analysis,phylogenetic tree construction,and estimation of differentiation times for SFGR were conducted in DNASP v6,IQ-tree v2.2.0,and BEAST v2.7.4 software.The obtained 446 tick samples belonged to three categories:(1)Haemaphy-salis spp.,including Haemaphysalis qinghaiensis(n=192)and H.danieli(n=37);(2)Dermacentor spp.,including Dermacentor ever-estianus(n=121),D.nuttalli(n=55),and D.silvarum(n=36);and(3)Hyalomma marginatum(n=5).Rickettsia raoultii was de-tected in D.everestianus,D.silvarum,D.nuttalli,H.qinghaiensis,and H.danieli,with infection rates of 95.9%,80.6%,69.1%,4.1%,and 2.7%,respectively.R.sibirica subsp.sibirica BJ-90 was found only in D.silvarum and D.nuttalli,with infection rates of 5.6%and 1.8%,respectively.The Candidatus R.gannanii F107 was found in H.danieli and H.qinghaiensis,with infection rates of 16.2%and 7.8%,respectively.Ca.R.hongyuanensis was detected only in H.qinghaiensis,with a prevalence of 16.3%.The prevalence of R.aeschlimannii was 20%and 2.7%in Hy.marginatum and H.danieli,respectively.Haplotype and nucleotide polymorphism analy-ses revealed 13 haplotypes in R.raoultii,with haplotype H13 as the dominant haplotype(42/192);seven haplotypes in Ca.R.ganna-nii F107,with haplotype H4 as the dominant haplotype(4/18);and three haplotypes in Ca.R.hongyuanensis,with haplotype H1 as the dominant haplotype(11/13).The phylogenetic tree indicated that the sequences of R.raoultii in selected areas of Qinghai and R.rhipicephali clustered into one branch;Ca.R.hongyuanensis and Ca.R.gannanii F107 clustered into one branch;and R.sibirica subsp.sibirica BJ-90 clustered into one branch with R.sibirica.Estimates of differentiation time revealed that the mean differentiation time for the six Rickettsia was approximately 2 000 Mya(95%CI:1 999.08-2 001.02 Mya).The tick species distributed in selected ar-eas of Qinghai are diverse,and this study provides the first report of Hy.marginatum in Qinghai Province.SFGR significantly varied in prevalence among tick species and showed high genetic diversity.

The study was aimed at identifying the diversity of tick species in selected areas of Qinghai,to analyze the genetic differentiation characteristics of tick-borne spotted fever group rickettsiae(SFGR),and to provide the theoretical basis for SFGR prevention and control in the region.The 16S rRNA gene was used for molecular biological identification of 446 collected tick samples,and the infection characteristics of SFGR in tick samples were determined according to the SFGR outer membrane protein A(ompA)gene.Haplotype analysis,phylogenetic tree construction,and estimation of differentiation times for SFGR were conducted in DNASP v6,IQ-tree v2.2.0,and BEAST v2.7.4 software.The obtained 446 tick samples belonged to three categories:(1)Haemaphy-salis spp.,including Haemaphysalis qinghaiensis(n=192)and H.danieli(n=37);(2)Dermacentor spp.,including Dermacentor ever-estianus(n=121),D.nuttalli(n=55),and D.silvarum(n=36);and(3)Hyalomma marginatum(n=5).Rickettsia raoultii was de-tected in D.everestianus,D.silvarum,D.nuttalli,H.qinghaiensis,and H.danieli,with infection rates of 95.9%,80.6%,69.1%,4.1%,and 2.7%,respectively.R.sibirica subsp.sibirica BJ-90 was found only in D.silvarum and D.nuttalli,with infection rates of 5.6%and 1.8%,respectively.The Candidatus R.gannanii F107 was found in H.danieli and H.qinghaiensis,with infection rates of 16.2%and 7.8%,respectively.Ca.R.hongyuanensis was detected only in H.qinghaiensis,with a prevalence of 16.3%.The prevalence of R.aeschlimannii was 20%and 2.7%in Hy.marginatum and H.danieli,respectively.Haplotype and nucleotide polymorphism analy-ses revealed 13 haplotypes in R.raoultii,with haplotype H13 as the dominant haplotype(42/192);seven haplotypes in Ca.R.ganna-nii F107,with haplotype H4 as the dominant haplotype(4/18);and three haplotypes in Ca.R.hongyuanensis,with haplotype H1 as the dominant haplotype(11/13).The phylogenetic tree indicated that the sequences of R.raoultii in selected areas of Qinghai and R.rhipicephali clustered into one branch;Ca.R.hongyuanensis and Ca.R.gannanii F107 clustered into one branch;and R.sibirica subsp.sibirica BJ-90 clustered into one branch with R.sibirica.Estimates of differentiation time revealed that the mean differentiation time for the six Rickettsia was approximately 2 000 Mya(95%CI:1 999.08-2 001.02 Mya).The tick species distributed in selected ar-eas of Qinghai are diverse,and this study provides the first report of Hy.marginatum in Qinghai Province.SFGR significantly varied in prevalence among tick species and showed high genetic diversity.

ObjectiveTo decipher the mechanism of Wenxiao powder in alleviating corticosterone-induced depression-like behaviors in mice. MethodMale ICR mice were randomized into normal， model， paroxetine （20 mg·kg^-1）， and low- and high-dose （3.27， 6.54 g·kg^-1， respectively） Wenxiao powder groups. The mice in normal and model groups received equal volume of saline. Other groups except the normal group were injected with corticosterone subcutaneously 0.5 h after gavage to induce depression. Mice were tested for depression-like behaviors after drug administration. Enzyme-linked immunosorbent assay （ELISA） was performed to measure the corticosterone content in the serum. Nissl staining was performed to observe the damage of hippocampal neurons. Immunofluorescence staining was employed to observe the expression of double cortin （DCX） in the dentate gyrus （DG） of the hippocampus. Western blot was employed to determine the expression of proteins in the brain-derived neurotrophic factor （BDNF）/tyrosine kinase receptor B （TrkB）/extracellular signal-regulated kinase （ERK）/cAMP-response element-binding protein （CREB） pathway in the hippocampus. ResultCompared with the normal group， the model group showed decreased sucrose preference rate， increased immobility time in the tail suspension test （P<0.01）， and reduced residence time in the central area of the open field and the total movement distance （P<0.05， P<0.01）. In addition， the modeling elevated the corticosterone level in the serum （P<0.01）， decreased the volume and intensified the nuclear staining of hippocampal neurons in the DG area， reduced the expression of DCX in the DG area， and down-regulated the protein levels of BDNF， phosphorylated （p）-TrkB， p-ERK， and p-CREB in the hippocampus （P<0.05， P<0.01）. Compared with the model group， low-dose Wenxiao powder improved the mouse behavivors in the sucrose preference， open field， and tail suspension tests （P<0.05， P<0.01）， and high-dose Wenxiao powder improved the behaviors in the sucrose preference and open field tests （P<0.05， P<0.01）. In addition， Wenxiao powder lowered the serum corticosterone level （P<0.01） and recovered the structure and morphology of neurons with obvious nuclei and presence of Nissl bodies in the DG area of the hippocampus. Moreover， Wenxiao powder at both doses promoted the expression of DCX in the DG area， and high-dose Wenxiao powder up-regulated the protein levels of BDNF， p-TrkB， p-ERK， and p-CREB in the hippocampus （P<0.05， P<0.01）. ConclusionWenxiao powder can alleviate corticosterone-induced depression-like behaviors and promote neurogenesis in mice possibly by activating the BDNF/TrkB/ERK/CREB signaling pathway.

Background Excessive radon exposure is considered the second risk factor for lung cancer. Since the opening of the subway in a city of Zhejiang Province, the exposure level of radioactive gas radon in subway stations and its impact on occupational health have become one of the important issues of public concern. Objective To monitor the radon concentration of subways in a city in Zhejiang Province and explore the effect of radon exposure on chromosome aberration and micronuclei in the working population. Methods A total of 55 vehicle control rooms of 55 stations affiliated to two different subway lines in a city were measured for one year; the 110 ticket offices and 55 security checkpoints from the same 55 stations were measured from 16 March to 14 June. The radon concentrations were compared by job types, subway lines, and seasons referring to Measurement methods for determination of radon in environmental air (HJ 1212-2021). Peripheral blood lymphocyte chromosome aberration and micronucleus analyses were conducted in 165 subway workers from monitoring sites for three different job types, then the influencing factors were analyzed. The detection methods were adopted from the standards of Test and assessment of chromosomal aberrations on occupational health examinations for radiation workers (GBZ/T 248-2014) and Standard for the method of micronucleus detection in lymphocytes on occupational health examination for radiation workers and exposure dose estimation (GBZ/T 328-2023). Results The radon concentration range of the target subways in Zhejiang Province was 10-320 Bq·m⁻³, all lower than the national limit (≤400 Bq·m⁻³). The differences in radon radioactivity levels among different lines, job types, and time segments were statistically significant (P<0.05). The rates of chromosomal aberration and micronucleus formation among the 165 subjects were 0.224% and 0.024%, respectively. There were significant differences in the rates of chromosome aberration and micronuclei among different jobs (vehicle control room, ticket office, security checkpoint) (P<0.05), but the abnormal rates were lower than the limits of the corresponding national standard. No significant correlation was found between jobs and chromosomal aberrations or micronuclei (P>0.05). Chromosome aberration and micronuclei varied by age, subway station seniority, and smoking (P<0.05). No effect of the above factors on chromosome aberration and micronuclei was observed by logistic regression (P>0.05). Conclusion The radon concentration in the target subway system is at a normal level. The rates of chromosomal aberration and micronucleus formation vary by jobs, but both are lower than the corresponding national limits. Therefore, radon exposure has not yet caused outstanding health impact on the subway workers.

AIM To study the secondary metabolites from endophytic fungi of Taxus wallichiana var.chinensis (Pilger) Florin and their biological activities.METHODS Endophytes were isolated and purified by tissue block and scribing method,the obtained strains were identified by molecular method.Macroporous resin column and silica gel column were used for separation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The inhibition of phosphatidylinositol 3-kinase,phosphodiesterase and fusarium oxysporum activities were determined by chemiluminescence kinase assay kit method,resazurin cell viability assay and enzyme linked immunosorbent assay.RESULTS Eight endophytic fungi were isolated and identified as Trichoderma,Aspergillus,Gongronella,Phoma,Penicillium.Strain ZGG-5 had strong PI3K inhibitory activity with IC50 value of (0.17±0.05)μg/mL,and was identified as Gongronella butleri.Thirteen compounds were isolated from ZGG-5,and compound 13 had strong activities of inhibiting PI3K at concentration of 1 mg/mL,and the inhibitory rate was (97.8±0.51)％.CONCLUSION T.wallichiana var.chinensis contains endophytic fungi with strong activities of inhibiting PI3K.Compounds 1-12 are first isolated from G.butleri.

AIM To study the secondary metabolites from endophytic fungi of Taxus wallichiana var.chinensis (Pilger) Florin and their biological activities.METHODS Endophytes were isolated and purified by tissue block and scribing method,the obtained strains were identified by molecular method.Macroporous resin column and silica gel column were used for separation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The inhibition of phosphatidylinositol 3-kinase,phosphodiesterase and fusarium oxysporum activities were determined by chemiluminescence kinase assay kit method,resazurin cell viability assay and enzyme linked immunosorbent assay.RESULTS Eight endophytic fungi were isolated and identified as Trichoderma,Aspergillus,Gongronella,Phoma,Penicillium.Strain ZGG-5 had strong PI3K inhibitory activity with IC50 value of (0.17±0.05)μg/mL,and was identified as Gongronella butleri.Thirteen compounds were isolated from ZGG-5,and compound 13 had strong activities of inhibiting PI3K at concentration of 1 mg/mL,and the inhibitory rate was (97.8±0.51)％.CONCLUSION T.wallichiana var.chinensis contains endophytic fungi with strong activities of inhibiting PI3K.Compounds 1-12 are first isolated from G.butleri.

COVID-19 is caused by coronavirus SARS-CoV-2. Current systemic vaccines generally provide limited protection against viral replication and shedding within the airway. Recombinant VSV (rVSV) is an effective vector which inducing potent and comprehensive immunities. Currently, there are two clinical trials investigating COVID-19 vaccines based on VSV vectors. These vaccines were developed with spike protein of WA1 which administrated intramuscularly. Although intranasal route is ideal for activating mucosal immunity with VSV vector, safety is of concern. Thus, a highly attenuated rVSV with three amino acids mutations in matrix protein (VSV_MT) was developed to construct safe mucosal vaccines against multiple SARS-CoV-2 variants of concern. It demonstrated that spike protein mutant lacking 21 amino acids in its cytoplasmic domain could rescue rVSV efficiently. VSV_MT indicated improved safeness compared with wild-type VSV as the vector encoding SARS-CoV-2 spike protein. With a single-dosed intranasal inoculation of rVSV_ΔGMT-S_Δ21, potent SARS-CoV-2 specific neutralization antibodies could be stimulated in animals, particularly in term of mucosal and cellular immunity. Strikingly, the chimeric VSV encoding S_Δ21 of Delta-variant can induce more potent immune responses compared with those encoding S_Δ21 of Omicron- or WA1-strain. VSV_MT is a promising platform to develop a mucosal vaccine for countering COVID-19.

Objective:To explore the application value of computed tomography (CT) plain scan and dynamic enhanced scan in the diagnosis of solitary pulmonary nodules.Methods:The clinical data of 120 patients with solitary pulmonary nodules detected by physical examination in Baoding First Central Hospital from January 2018 to December 2020 were retrospectively reviewed. All patients were confirmed by surgery and pathology, including 77 benign lesions and 43 malignant lesions; All patients underwent CT plain scan and dynamic enhanced scan before operation. The accuracy of the two examination methods in the diagnosis of benign and malignant lesions of solitary pulmonary nodules was analyzed and compared. The detection rate of CT dynamic enhanced scan imaging characteristics (vacuole sign, ground glass sign, spinous sign, lobulation sign, hair prick sign, blood vessel cluster, pleural depression) of benign and malignant lesions of solitary pulmonary nodules was compared, and the diagnostic value of CT plain scan and dynamic enhanced scan in the differential diagnosis of benign and malignant solitary pulmonary nodules was evaluated based on the results of surgical pathological diagnosis. The manifestations and characteristic curves of CT dynamic enhanced scan of solitary pulmonary nodules was analyzed.Results:The diagnostic accuracy of CT dynamic enhanced scan for solitary pulmonary nodules was 80.00% (96/120), which was higher than that of CT plain scan (63.33%) (76/120) ( P<0.05). The sensitivity, specificity, and negative predictive value of CT dynamic enhanced scan for the diagnosis of benign and malignant lesions of solitary pulmonary nodules were higher than those of CT plain scan (all P<0.05). Among the imaging characteristics of CT dynamic enhanced scans of malignant lesions, the ground glass sign, spinous process sign, lobulation sign, spiculation sign, vascular clustering and pleural indentation were detected more frequently than those of benign lesions (all P<0.05). Benign lesions usually showed homogeneous enhancement, and a few showed heterogeneous enhancement; Malignant nodules often showed uneven enhancement, and a few had even enhancement. The time density curves of dynamic enhanced CT values in the regions of interest of benign and malignant solitary pulmonary nodules were different. Conclusions:The value of dynamic enhanced CT scan in the differential diagnosis of benign and malignant lesions of solitary pulmonary nodules is higher than that of CT plain scan, and the imaging features are obvious, with higher sensitivity and specificity, which is worthy of application.

Chronic myeloid leukemia (CML) is a slowly progressing hematopoietic cell disorder. Sphingosine kinase 1 (SPHK1) plays established roles in tumor initiation, progression, and chemotherapy resistance in a wide range of cancers, including leukemia.However, small-molecule inhibitors targeting SPHK1 in CML still need to be developed. This study revealed the role of SPHK1 in CML and investigated the potential anti-leukemic activity of hirsuteine (HST), an indole alkaloid obtained from the oriental plant Uncaria rhynchophylla, in CML cells. These results suggest that SPHK1 is highly expressed in CML cells and that overexpression of SPHK1 represents poor clinical outcomes in CML patients. HST exposure led to G2/M phase arrest, cellular apoptosis, and downregulation of Cyclin B1 and CDC2 and cleavage of Caspase 3 and PARP in CML cells. HST shifted sphingolipid rheostat from sphingosine 1-phosphate (S1P) towards the ceramide coupled with a marked inhibition of SPHK1. Mechanistically, HST significantly blocked SPHK1/S1P/S1PR1 and BCR-ABL/PI3K/Akt pathways. In addition, HST can be docked with residues of SPHK1 and shifts the SPHK1 melting curve, indicating the potential protein-ligand interactions between SPHK1 and HST in both CML cells. SPHK1 overexpression impaired apoptosis and proliferation of CML cells induced by HST alone. These results suggest that HST, which may serve as a novel and specific SPHK1 inhibitor, exerts anti-leukemic activity by inhibiting the SPHK1/S1P/ S1PR1 and BCR-ABL/PI3K/Akt pathways in CML cells, thus conferring HST as a promising anti-leukemic drug for CML therapy in the future.

Objective:To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT).Methods:A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ 2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results:A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment.Conclusion:ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.