Tracheobronchial adenoid cystic carcinoma （TACC） is a low-grade malignant tumor originating from the airway mucosa. Despite its slow progression，it is characterized by high invasiveness，frequent recurrence，and a strong tendency for metastasis. Preclinical studies have shown that vascular-targeted therapy holds significant potential. However，an effective systemic treatment for TACC has not been established yet. This study explored TACC from the perspective of "Feiji" in traditional Chinese medicine （TCM） as the starting point. It deeply investigated the mechanisms of abnormal collaterals and tumor vascular recruitment and further elaborated on the theoretical connection between abnormal collaterals and tumor vascular recruitment. Firstly，collateral hyperactivity led to disordered and erratic pulmonary collaterals. Their abnormal structures were similar to the disorderly and tortuous nature of tumor （pseudo）angiogenesis. This resulted in imbalances in the functions of circulation，perfusion，and reverse injection of the pulmonary collaterals，and then led to unrestrained collateral dysfunction and the accumulation of pathogenic factors. Secondly，the remodeling of the extracellular matrix （ECM） and epithelial-mesenchymal transition （EMT） in TACC were critical processes in vascular co-option （VCO），representing the micro-level manifestation of the displacement of nutrient and defense. During this process，ECM remodeling made TACC cells more likely to hijack normal blood vessels，creating a complex vascular microenvironment conducive to tumor growth. In terms of treatment，this study proposed a TCM strategy of "regulating collaterals to expel pathogenic factors and nourishing collaterals to strengthen the healthy Qi"，and listed potential TCM. These were intended to regulate the Qi and blood in the collaterals，repair the functions of abnormal collaterals，and intervene in the vascular recruitment process of TACC. Future research should focus on improving the TCM clinical syndrome characteristics of TACC. Through modern molecular biology techniques，it is necessary to deeply analyze the micro-level pattern of vascular recruitment in TACC. This would enrich the understanding of the profound connection between abnormal collaterals and tumor vascular recruitment，providing empirical evidence for TCM-targeted therapies for vascular recruitment in TACC.

Glioma is difficult to treat due to the unique tumor microenvironment and blood-brain barrier. (13aS)-3-Hydroxyl-6,7-dimethoxyphenanthro[9,10-b] indolizidine (PF403), a phenanthroindolizidine alkaloid, has been identified as a promising therapeutic agent for the treatment of glioma. However, the anti-glioma mechanism of PF403 in vivo has not been conclusively verified and must be further elucidated. Hence, a strategy without chemical modification was applied to identify the target of PF403. In this study, we identified nicotinamide phosphoribosyl transferase (NAMPT) as the target of PF403 by using thermal proteome profiling (TPP). Moreover, microscale thermophoresis (MST), surface plasmon resonance (SPR), and isothermal titration calorimetry (ITC) experiments confirmed that NAMPT exhibits good affinity for PF403. Direct and indirect enzyme activity assays revealed that PF403 inhibited the catalytic activity of NAMPT, leading to a decrease in the concentration of nicotinamide adenine dinucleotide (NAD⁺) in U87 cells. X-ray diffraction and amino acid spot mutation experiments revealed that PF403 primarily relies on the formation of pi-pi interactions with residue Tyr188 to maintain binding with NAMPT (PDB code 8Y55). After NAMPT was knocked down with lentivirus, PF403 lost or partially lost its antitumor activity at the cellular and animal levels. These findings suggest that PF403 exerts antitumor activity by directly targeting NAMPT.

Objective:To provide data support for the improvement of aeromedical support by summarizing the disease characteristics and analyzing the change pattern of disease spectrum for the temporarily grounded military flying personnel assessed in the annual physical examination.Methods:The clinical data of military flying personnel who were assessed as temporary grounding in the annual physical examination in Air Force Healthcare Center for Special Services Hangzhou from 2012 to 2021 were retrospectively analyzed. They were divided into 2012—2016 group and 2017—2021 group with a 5-year span. The distributions of disease, age and aircraft type were statistically compared between 2 groups of temporarily grounded military flying personnel.Results:A total of 522 flying personnel, aged from 23-55 years and flew 100-20 000 h, were enrolled. There were 242 cases in 2012—2016 group and 280 cases in 2017—2021 group. Compared with 2012—2016 group, 2017—2021 group was found 9 new diseases. Among the top 10 diseases in 2017—2021 group, the proportion of pulmonary nodules, hypertension, diabetes, abnormal tumor indicators, positive fecal occult blood was increased, and the proportion of thyroid nodules, abnormal liver function, arrhythmia was decreased. There was a significant difference in the age distribution between 2 groups of temporarily grounded military flying personnel ( χ2=20.97, P<0.001). The proportion of flying personnel aged ≥40 years old was increased in 2017—2021 group. The proportion of fighter-trainer pilots and bomb-transporter flying personnel increased in 2017—2021 group, and the difference was significant ( χ2=14.66, P=0.001). Conclusions:In recent years, more diseases, mostly occult diseases, are found in the disease spectrum of the temporarily grounded military flying personnel. Since some insidious diseases require more time to observe their convalescent effect and the influence to flight but the routine phone follow-up is called within 2 months of discharge, so a long-term and effective follow-up mechanism is suggested.

Objective:To analyze the results of low-dose lung CT screening for military flying personnel, and to explore its influence on aeromedical identification.Methods:The imaging and aeromedical identification data of military flying personnel who underwent annual physical examination in Air Force Healthcare Center for Special Services Hangzhou were analyzed retrospectively. According to the imaging manifestations, the enrolled low-dose lung CT results were divided into the following 6 categories: no obvious abnormality, old lesions, pulmonary nodules, pulmonary bullae, pulmonary inflammatory lesions and extrapulmonary diseases. The detection rates of various examination results were statistically analyzed. The military flying personnel were divided into 20-29 years group, 30-39 years group, 40-49 years group and 50-59 years group, and they were divided into fighter pilots, bomb-trans-helicopter pilots, and bomb-trans-helicopter crew members according to the post. The differences in various examination results of flying personnel were compared among different age groups and post groups. The influence of examination results on the aeromedical identification results was analyzed in accordance with the 3-level conclusions as not affecting the conclusion of aeromedical identification, leading to the decline of health grade and leading to temporary grounding. The overall significance level α=0.05, and the pairwise comparison with P< α/statistical times was statistically significant. Results:A total of 2 274 flying personnel was enrolled. The detection rates of various examination results from high to low was no obvious abnormality (45.82%), old lesions (37.55%), pulmonary nodules (14.86%), pulmonary bullae (6.73%), pulmonary inflammatory lesions (2.59%) and extrapulmonary diseases (1.85%). There was significant difference among examination results of different age groups ( χ2=183.43, P<0.001). The detection rate of no obvious abnormality was the highest in 20-29 years group and 30-39 years group; the detection rate of old lesions was the highest in 40-49 years group and 50-59 years group. The detection rate of no obvious abnormality decreased with age, and the detection rate of old lesions increased with age (both P< α/6=0.008). The detection rate of pulmonary nodules in 40-49 years group was higher than that in 30-39 years group; the detection rate of pulmonary bullae in 50-59 years group was higher than that in the other age groups; the detection rates of pulmonary inflammatory lesions in 40-49 years group and 50-59 years groups were higher than those in the other age groups; the detection rate of extrapulmonary diseases in 50-59 years group was higher than that in 20-29 years group, and the differences were statistically significant (all P<0.008). There was significant difference in the results of flying personnel among different post groups ( χ2=33.78, P=0.001). The detection rate of no obvious abnormality in bomb-trans-helicopter crew members group was lower than that in bomb-trans-helicopter pilots group and fighter-trainer pilots group, but the detection rates of old lesions and pulmonary nodules were higher than those in the other groups, the differences were statistically significant (all P< α/3=0.017). The detection rate of pulmonary bullae in fighter-trainer pilots group was lower than that in bomb-trans-helicopter crew members group, and the difference was statistically significant ( P<0.017). Although majority of the detected results had no effect on the results of aeromedical identification, there was a significant difference in the proportion of health grade decline and temporary grounding caused by different detected diseases (Fisher exact test value was 82.81, P<0.001). Conclusions:Low-dose lung CT screening plays an important role in the physical examination and aeromedical identification for military flying personnel. It can be routinely used during the annual physical examination for flying personnel, and the aeromedical identification standards and strategies for lung related diseases should be updated synchronously.

Abstract: Objective To clarify the long-term evolution of hepatitis B virus (HBV) quasi-species in HBsAg asymptomatic carriers in Long'an county, Guangxi. Methods ELISA was used to detect serological markers of HBV. Viral loads were measured by real time PCR. HBV DNA was extracted from serum by kits. The whole HBV genome was amplified using nested PCR and amplicons were sequenced by next-generation sequencing (NGS). These sequences from NGS were analyzed by the software like Mega. Results Serum samples were collected from 9 HBsAg asymptomatic carriers in Longan County，Guangxi at 4 different time points in 2004, 2007, 2013, 2019 or 2020. A total of 23 serum samples and 309 full-length gene quasi-species sequences were obtained, with an average amount of (0.18±0.07) G sequencing data for each sample. Genotype of 55.54%(5/9) the studied subjects underwent genotype conversion during the long-term evolution process of HBV quasi-species, and the genotyping results of the phylogenetic tree in the PreS/S region are in perfect agreement with the results of the whole genome analysis; recombinant B/C, I/C were found; the Sn ranged from 0 to 0.37 and the genetic diversity ranged from 0 to 0.11, respectively. A total of 21 special single nucleotide/amino acid mutations (7 in the S region, 2 in the X region, 3 in the PreC region and 9 in the BCP region) and 6 deletion mutations were detected, multiple mutations were found and no drug resistant mutations were found; 77.8%(7/9) of the HBV strains carried by the subjects in 2004 had double mutations at nt1 762(A→T) and 1 764(G→A) and a stop mutation at nt1 896(G→A); HBV mutations can be restored from the mutant type to the wild type and (or) vice versa without antiviral drug pressure, and The evolution rate of HBV genome was 2.03×10-5~3.50×10-3.Conclusion HBV genotype, recombinants, genetic complexity and diversity of HBV quasi-species can change over time during in natural infection. The transformation between HBV mutation type and wild type reduces the value of predicting clinical outcomes by genetic types and related mutations to some extent. The HBV genome evolution rate of asymptomatic carriers of HBsAg in Long'an County is very high.

BACKGROUND: Colorectal carcinogenesis and progression are related to the gut microbiota and the tumor immune microenvironment. Our previous clinical trial demonstrated that berberine (BBR) hydrochloride might reduce the recurrence and canceration of colorectal adenoma (CRA). The present study aimed to further explore the mechanism of BBR in preventing colorectal cancer (CRC). METHODS: We performed metagenomics sequencing on fecal specimens obtained from the BBR intervention trial, and the differential bacteria before and after medication were validated using quantitative polymerase chain reaction. We further performed ApcMin/+ animal intervention tests, RNA sequencing, flow cytometry, immunohistochemistry, and enzyme-linked immunosorbent assays. RESULTS: The abundance of fecal Veillonella parvula ( V . parvula ) decreased significantly after BBR administration ( P = 0.0016) and increased through the development from CRA to CRC. Patients with CRC with a higher V. parvula abundance had worse tumor staging and a higher lymph node metastasis rate. The intestinal immune pathway of Immunoglobulin A production was activated, and the expression of TNFSF13B (Tumor necrosis factor superfamily 13b, encoding B lymphocyte stimulator [BLyS]), the representative gene of this pathway, and the genes encoding its receptors (interleukin-10 and transforming growth factor beta) were significantly upregulated. Animal experiments revealed that V. parvula promoted colorectal carcinogenesis and increased BLyS levels, while BBR reversed this effect. CONCLUSION: BBR might inhibit V. parvula and further weaken the immunomodulatory effect of B cells induced by V. parvula , thereby blocking the development of colorectal tumors. TRIAL REGISTRAION ClinicalTrials.gov, No. NCT02226185.
Animals ; Humans ; Berberine/therapeutic use* ; Carcinogenesis ; Veillonella ; Colorectal Neoplasms/genetics* ; Tumor Microenvironment

Objective:To provide data support for the improvement of aeromedical support by summarizing the disease characteristics and analyzing the change pattern of disease spectrum for the temporarily grounded military flying personnel assessed in the annual physical examination.Methods:The clinical data of military flying personnel who were assessed as temporary grounding in the annual physical examination in Air Force Healthcare Center for Special Services Hangzhou from 2012 to 2021 were retrospectively analyzed. They were divided into 2012—2016 group and 2017—2021 group with a 5-year span. The distributions of disease, age and aircraft type were statistically compared between 2 groups of temporarily grounded military flying personnel.Results:A total of 522 flying personnel, aged from 23-55 years and flew 100-20 000 h, were enrolled. There were 242 cases in 2012—2016 group and 280 cases in 2017—2021 group. Compared with 2012—2016 group, 2017—2021 group was found 9 new diseases. Among the top 10 diseases in 2017—2021 group, the proportion of pulmonary nodules, hypertension, diabetes, abnormal tumor indicators, positive fecal occult blood was increased, and the proportion of thyroid nodules, abnormal liver function, arrhythmia was decreased. There was a significant difference in the age distribution between 2 groups of temporarily grounded military flying personnel ( χ2=20.97, P<0.001). The proportion of flying personnel aged ≥40 years old was increased in 2017—2021 group. The proportion of fighter-trainer pilots and bomb-transporter flying personnel increased in 2017—2021 group, and the difference was significant ( χ2=14.66, P=0.001). Conclusions:In recent years, more diseases, mostly occult diseases, are found in the disease spectrum of the temporarily grounded military flying personnel. Since some insidious diseases require more time to observe their convalescent effect and the influence to flight but the routine phone follow-up is called within 2 months of discharge, so a long-term and effective follow-up mechanism is suggested.

Objective:To analyze the results of low-dose lung CT screening for military flying personnel, and to explore its influence on aeromedical identification.Methods:The imaging and aeromedical identification data of military flying personnel who underwent annual physical examination in Air Force Healthcare Center for Special Services Hangzhou were analyzed retrospectively. According to the imaging manifestations, the enrolled low-dose lung CT results were divided into the following 6 categories: no obvious abnormality, old lesions, pulmonary nodules, pulmonary bullae, pulmonary inflammatory lesions and extrapulmonary diseases. The detection rates of various examination results were statistically analyzed. The military flying personnel were divided into 20-29 years group, 30-39 years group, 40-49 years group and 50-59 years group, and they were divided into fighter pilots, bomb-trans-helicopter pilots, and bomb-trans-helicopter crew members according to the post. The differences in various examination results of flying personnel were compared among different age groups and post groups. The influence of examination results on the aeromedical identification results was analyzed in accordance with the 3-level conclusions as not affecting the conclusion of aeromedical identification, leading to the decline of health grade and leading to temporary grounding. The overall significance level α=0.05, and the pairwise comparison with P< α/statistical times was statistically significant. Results:A total of 2 274 flying personnel was enrolled. The detection rates of various examination results from high to low was no obvious abnormality (45.82%), old lesions (37.55%), pulmonary nodules (14.86%), pulmonary bullae (6.73%), pulmonary inflammatory lesions (2.59%) and extrapulmonary diseases (1.85%). There was significant difference among examination results of different age groups ( χ2=183.43, P<0.001). The detection rate of no obvious abnormality was the highest in 20-29 years group and 30-39 years group; the detection rate of old lesions was the highest in 40-49 years group and 50-59 years group. The detection rate of no obvious abnormality decreased with age, and the detection rate of old lesions increased with age (both P< α/6=0.008). The detection rate of pulmonary nodules in 40-49 years group was higher than that in 30-39 years group; the detection rate of pulmonary bullae in 50-59 years group was higher than that in the other age groups; the detection rates of pulmonary inflammatory lesions in 40-49 years group and 50-59 years groups were higher than those in the other age groups; the detection rate of extrapulmonary diseases in 50-59 years group was higher than that in 20-29 years group, and the differences were statistically significant (all P<0.008). There was significant difference in the results of flying personnel among different post groups ( χ2=33.78, P=0.001). The detection rate of no obvious abnormality in bomb-trans-helicopter crew members group was lower than that in bomb-trans-helicopter pilots group and fighter-trainer pilots group, but the detection rates of old lesions and pulmonary nodules were higher than those in the other groups, the differences were statistically significant (all P< α/3=0.017). The detection rate of pulmonary bullae in fighter-trainer pilots group was lower than that in bomb-trans-helicopter crew members group, and the difference was statistically significant ( P<0.017). Although majority of the detected results had no effect on the results of aeromedical identification, there was a significant difference in the proportion of health grade decline and temporary grounding caused by different detected diseases (Fisher exact test value was 82.81, P<0.001). Conclusions:Low-dose lung CT screening plays an important role in the physical examination and aeromedical identification for military flying personnel. It can be routinely used during the annual physical examination for flying personnel, and the aeromedical identification standards and strategies for lung related diseases should be updated synchronously.

Objective:To investigate the immune effects of Clostridium difficile toxoid B (CdtB) vaccine formulated with different mucosal adjuvants through microneedle immunization, and to provide ideas for the prevention and treatment of Clostridium difficile infection. Methods:CdtB vaccine was prepared with purified Clostridium difficile toxin B(TcdB) after formaldehyde detoxification. Female BALB/c mice were immunized with different doses of vaccine alone or in combination with mucosal adjuvants. The titers of specific serum IgG and fecal IgA were detected at 0 d, 7 d, 14 d, 28 d and 42 d after immunization. The protective effects of CdtB vaccine were evaluated by cell neutralization assay and Clostridium difficile challenge infection. Results:(1) With the increase of immune dose, the mice immunized with CdtB vaccine alone by microneedle not only produced better serum specific IgG, but also had higher level of IgA in feces. (2) When the mice were immunized with CdtB vaccine containing LT or CTB adjuvant by microneedle, the trend of serum specific IgG titer in each group increased with the increase of immune dose, especially in the group containing LT adjuvant. There were significant differences in the trend of specific IgA titer in feces between the adjuvant groups and the group without adjuvant, but the adjuvant effect was not obvious. (3) No significant difference in serum IgG titer was observed between the mice immunized with 10 μg CdtB by microneedle or intraperitoneal injection, but microneedle immunization significantly increased fecal IgA level. (4) The neutralization titers of specific antibodies in mouse serum after immunization and the test results of challenge protection in mice confirmed that the use of CdtB vaccine had certain protective effects.Conclusions:CdtB vaccine had better immune effects in mice through microneedle immunization, but the adjuvant effects of LT and CTB were not significant.

Objective:To explore the association of DNA methylation with immune response to hepatitis B (HepB) vaccine in Han nationality children from Guangxi province.Methods:A total of 263 children aged 8-9 months who had completed HepB immunization program were recruited from three hospitals in Guangxi province by using unmatched case-control method. Children with the HepB surface antibody concentration(Anti -HBs)<100 mIU/ml was set as the case group and ≥100 mIU/ml as the control group. Multiplex PCR and heavy sulfite sequencing were used to treat the samples. Illumina platform was used for high-throughput DNA methylation sequencing of IFNG gene target regions and CpG sites. Unconditional logistic regression was used to analyze the association between cytosine-phospho-guanosine DNA methylation at 18 loci of IFNG gene and HepB immune response level. Results:There were 104 children in the case group and 159 in the control group. The median ( Q1, Q3) level of anti -HBs in two groups were 62.34 (30.06, 98.88) mIU/ml and 1 089.10 (710.35, 1 233.45) mIU/ml. The methylation levels of IFNG_1 gene 44 and 93 locus in the case group were higher than those in the control group ( P<0.05). The unconditional logistic regression model showed that the DNA methylation level of IFNG_1 gene at 44 ( OR=1.18, 95% CI: 1.03-1.35) and 93 ( OR=1.21, 95% CI: 1.07-1.38) locus was associated with the HepB response level. Conclusion:The changes of DNA methylation at locus 44 and 93 of IFNG_1 gene may be relevant factors affecting the response level of HepB in Han nationality children from Guangxi province.