[Genistein loaded by PRF improved bone healing in obese mice].

PURPOSE: To clarify the effect of genistein(GEN) on osteogenic differentiation and explore the effect of GEN loaded by platelet-rich fibrin (PRF) on the repair process of bone defects in obese mice. METHODS: In in vitro experiments, the effect of GEN(0, 0.1, 1, 10, 50 µmol/L) on the proliferation of mouse embryonic osteoblast precursor cells (MC3T3-E1) was determined by CCK 8. Alkaline phosphatase(ALP) staining and quantitative detection of ALP activity were performed to determine the changes of ALP activity in cells; RNA and protein expression levels of ALP, osteopontin (OPN) and osteocalcin (OCN) were detected by quantitative real-time PCR(qRT-PCR) and Western blot. Alizarin red staining was used to define the effect of GEN on mineralization of MC3T3-E1. To verify the feasibility of the PRF drug loading, the ultrastructure of PRF was subsequently observed under SEM. In in vivo experiments, obese C57 mouse models were established by high-fat diet feeding. On this basis, skull defect models with a diameter of 2.8 mm were established, and the prepared GEN/PRF complexes were placed into the bone defect area. The effects of GEN on skull defect repair in obese mice were evaluated by Micro-CT scanning and hematoxylin-eosin(H-E) staining. Statistical analysis was performed with GraphPad Prism 5.0 software package. RESULTS: CCK 8 results showed that 0.1, 1 µmol/L GEN promoted cell proliferation within 7 days(P＜0.05); 10 µmol/L GEN had no significant effect on the process of cell proliferation. From the second day, 50 µmol/L GEN significantly inhibited cell growth and showed cytotoxicity(P＜0.05). These two concentrations had similar effects in promoting cellular osteogenic differentiation. SEM results showed that PRF presented a 3-dimensional network structure, providing space for loading drug molecules. In in vivo experiments, the body weight of mice in the high-fat diet (HFD) group was 27.7% greater than that in the normal diet group(P＜0.05) and had abnormal glucose tolerance (P＜0.05). Micro-CT showed that compared with the normal diet group, the number of bone trabeculae in the femur of obese mice was decreased(P＜0.05), the distance between bone trabeculae was widened(P＜0.05), and the bone density was decreased (P＜0.05). In addition, GEN (0.1, 1.0 µmol/L) loaded by PRF increased bone volume fraction in the skull of obese mice (P＜0.05). H-E results showed that GEN/PRF promoted the healing of the bone defects. CONCLUSIONS: GEN promotes osteogenic differentiation of MC3T3-E1, and it can effectively accelerate the healing of cranial bone defects after loading with PRF in obese mice.

Cell-type specific circadian transcription factor BMAL1 roles in excitotoxic hippocampal lesions to enhance neurogenesis.

Circadian clocks, generating daily rhythms in biological processes, maintain homeostasis in physiology, so clock alterations are considered detrimental. Studies in brain pathology support this by reporting abnormal circadian phenotypes in patients, but restoring the abnormalities by light therapy shows no dramatic effects. Recent studies on glial clocks report the complex effects of altered clocks by showing their beneficial effects on brain repairs. However, how neuronal clocks respond to brain pathology is elusive. This study shows that neuronal BMAL1, a core of circadian clocks, reduces its expression levels in neurodegenerative excitotoxicity. In the dentate gyrus of excitotoxic hippocampal lesions, reduced BMAL1 in granule cells precedes apoptosis. This subsequently reduces BMAL1 levels in neighbor neural stem cells and progenitors in the subgranular zone, enhancing proliferation. This shows the various BMAL1 roles depending on cell types, and its alterations can benefit brain repair. Thus, cell-type-specific BMAL1 targeting is necessary to treat brain pathology.

Postoperative pulmonary complications in patients undergoing aortic surgery: A single-center retrospective study.

Postoperative pulmonary complications (PPCs) are among the most common complications after cardiovascular surgery. This study aimed to explore the real incidence of and risk factors for PPC in patients with acute type A aortic dissection (ATAAD) who underwent total aortic arch replacement combined with the frozen elephant trunk (TAR + FET). In total, 305 ATAAD patients undergoing TAR + FET from January 2021 to August 2022 in a single-center were divided into PPCs or non-PPCs group. The incidence of PPCs was calculated, risk factors of PPCs were analyzed, and postoperative outcomes were compared between these 2 groups. The incidence of any PPC was 29.2%. And the incidence of respiratory infection, respiratory failure, pleural effusion, atelectasis, pneumothorax, acute respiratory distress syndrome, aspiration pneumonitis, pulmonary edema and bronchospasm was 23.0%, 12.5%, 10.5%, 1.0%, 0.7%, 1.0%, 0%, 0.7%, 0%, respectively. The logistic regression analysis revealed that the history of diabetes, history of renal dysfunction, preoperative SpO2 <90%, cardiopulmonary bypass duration, fresh frozen plasma volume and platelet concentrates volume were independent risk factors for PPCs. Among 2 groups, postoperative ventilation duration, postoperative length of stay in intensive care unit and hospital were (73.5 ± 79.0 vs 24.8 ± 35.2 hours; P < .001), (228.3 ± 151.2 vs 95.2 ± 72.0 hours; P < .001) and (17.9 ± 8.8 vs 11.5 ± 6.2 days; P < .001). There was no difference between 2 groups of in-hospital mortality rate. Additionally, other short-term outcomes were also significantly poorer in patients with PPCs. PPCs are common in ATAAD patients undergoing TAR + FET, and could be multifactorial. PPCs occurrence are associated with poor patient outcomes postoperatively and worth further investigation.

Effect of dominant vertebral artery angle on basilar artery curvature and plaque.

Background: Quantification of vertebral arteries can provide insights into basilar curvature and plaque. Therefore, this retrospective study aimed at identifying the dominant vertebral artery (VA) causing basilar artery (BA) curvature and to further quantify the effect of dominant VA angle on BA curvature and BA plaque using high-resolution magnetic resonance imaging (HRMRI) and 3-dimensional time-of-flight magnetic resonance angiography (3D-TOF-MRA). Methods: This retrospective analysis included 521 participants who underwent HRMRI in the China-Japan Friendship Hospital from November 2015 to October 2021 for neurological symptoms or signs. The VA angle more related to BA curvature was defined as the dominant VA angle. Multivariable linear regression analysis was used to evaluate the relationship between the dominant VA angle and mid-BA angle, while multivariable logistics regression was used to evaluate the influence of the dominant VA angle and clinical risk factors on BA plaque. Results: In total, 259 participants were included in this study (mean age 53.71±13.12 years; 146 males). The balanced-type participants had a significantly lower probability of BA plaques (P<0.001). The Chi-squared test showed that the BA curvature direction was significantly associated with the side with larger VA diameter (P<0.001) and larger VA angle (P<0.001). As a result, the VA angle on the side with the larger diameter or the larger VA angle when the diameters were similar was considered to be the dominant VA angle. The dominant VA angle was independently correlated with the mid-BA angle (P<0.001). In addition, the dominant VA angle was also an independent risk factor for BA plaque. Additionally, 80° was the cutoff value of the dominant VA angle, and when the dominant VA angle was greater than 80°, the risk of BA plaque increased about 18-fold [odds ratio, 18.951; 95% confidence interval (CI): 4.545-79.026; P<0.001]. Conclusions: The dominant VA angle was independently associated with BA plaque, and a dominant VA angle greater than 80° may be a marker for a high risk of posterior circulation atherosclerosis.

Target-triggered 'colorimetric-fluorescence' dual-signal sensing system based on the versatility of MnO2 nanosheets for rapid detection of uric acid.

A simple dual-signal assay that combined colorimetric and fluorometric strategy for uric acid (UA) rapid detection was designed based on the versatility of facile synthesized MnO2 nanosheet. The oxidization of 3,3',5,5'-tetramethylbenzidine (TMB) and the fluorescence quenching of quantum dots (QDs) occurred simultaneously in the presence of MnO2 nanosheet. UA could decompose MnO2 nanosheet into Mn2+, resulting in the fluorescence recovery of QDs, along with the fading of the blue color of ox TMB. Based on the principles above, the detection of UA could be realized by the change of the dual signals (colorimetric and fluorometric). The linear range of the colorimetric mode was 5-60 µmol L-1, and the limit of detection (LOD) was 2.65 µmol L-1; the linear range of the fluorescence mode was wide at 5-120 µmol L-1, and the LOD could be as low as 1.33 µmol L-1. The method was successfully used for analyzing UA levels in human serum samples, indicating that this new dual-signal method could be applied in clinical diagnosis.

Digital-filter-aided crosstalk-mitigation for a high spatial resolution AWGR-based 2D IR beam-steered indoor optical wireless communication system.

The growing demand for wireless connectivity is attracting interest in optical wireless communication (OWC) technique. In this paper, a filter-aided crosstalk mitigation scheme, employing digital Nyquist filters, is proposed to eliminate the trade-off between the spatial resolution and the channel capacity for the AWGR-based 2D infrared beam-steered indoor OWC system. By shaping the transmitted signal for narrow spectral occupancy, the inter-channel crosstalk resulting from the imperfect AWGR filtering can be avoided, which enables a denser AWGR grid. In addition, the spectral-efficient signal reduces the bandwidth requirement of the AWGR, which allows a low-complexity AWGR design. Thirdly, the proposed method is not sensitive to the wavelength misalignment between AWGRs and lasers, which relaxes the design of high wavelength stability lasers. Moreover, the proposed method is cost-efficient as we can make use of the mature DSP technique without additional optical components. The 20-Gbit/s data rate OWC capacity using PAM4 format has been experimentally demonstrated over a 6-GHz bandwidth-limited AWGR-based 1.1-m free-space link. The experimental results show the feasibility and effectiveness of the proposed method. By combining our proposed method with the polarization orthogonality technique, a promising capacity per beam of 40 Gbit/s is potentially attainable.

Effect of Mid-Basilar Artery Angle and Plaque Characteristics on Pontine Infarction in Patients with Basilar Artery Plaque.

AIMS: The basilar artery (BA) geometry and plaque characteristics may play an important role in the development of atherosclerosis. This study was performed to explore the relationship between the mid-BA angle and plaque characteristics and its effect on pontine infarction using high-resolution magnetic resonance imaging and three-dimensional time-of-flight magnetic resonance angiography. METHODS: In total, 77 patients with BA plaques were included in this study. According to the presence of acute pontine infarction on diffusion-weighted imaging, the patients were divided into a pontine infarction group and pontine non-infarction group. The mid-BA angle, plaque burden, stenosis ratio, positive remodeling, and intraplaque hemorrhage were evaluated to investigate their effects on stroke. RESULTS: The pontine infarction group had a greater plaque burden, stenosis ratio, positive remodeling, and mid-BA angle than the pontine non-infarction group. The correlation between the plaque burden and mid-BA angle was the highest (r=0.441, P＜0.001). Multivariate logistic regression analysis showed that the plaque burden (odds ratio, 1.164; 95% confidence interval, 1.093-1.241; P＜0.001) was an independent risk factor for pontine infarction. CONCLUSION: The mid-BA angle may increase the incidence of pontine infarction by increasing the plaque burden.

Dynamics of a multi-strain malaria model with diffusion in a periodic environment.

This paper mainly explores the complex impacts of spatial heterogeneity, vector-bias effect, multiple strains, temperature-dependent extrinsic incubation period (EIP) and seasonality on malaria transmission. We propose a multi-strain malaria transmission model with diffusion and periodic delays and define the reproduction numbers Ri and R^i (i = 1, 2). Quantitative analysis indicates that the disease-free ω-periodic solution is globally attractive when Ri<1, while if Ri>1>Rj (i≠j,i,j=1,2), then strain i persists and strain j dies out. More interestingly, when R1 and R2 are greater than 1, the competitive exclusion of the two strains also occurs. Additionally, in a heterogeneous environment, the coexistence conditions of the two strains are R^1>1 and R^2>1. Numerical simulations verify the analytical results and reveal that ignoring vector-bias effect or seasonality when studying malaria transmission will underestimate the risk of disease transmission.

Role of ROS­mediated autophagy in melanoma (Review).

Melanoma is the most aggressive form of skin cancer with the poorest prognosis and its pathogenesis has yet to be fully elucidated. As key factors that regulate cellular homeostasis, both reactive oxygen species (ROS) and autophagy are involved in the development of melanoma, from melanomagenesis to progression and drug resistance. However, the interaction between ROS and autophagy in the etiology and treatment of melanoma is not well characterized. The present review examined the production of ROS and the role of oxidative stress in melanoma, and summarized the role of ROS­mediated autophagy in melanomagenesis and melanoma cell fate decision following treatment with various anticancer drugs. The present findings may lead to a better understanding of the pathogenesis and progression of melanoma, and suggest promising treatment options for this disease.

Association Between Some Different Obesity Anthropometric Indices and Type 2 Diabetes Mellitus in Middle-Aged and Elderly Chinese Men and Women in Beijing, China: A Cross-Sectional Study.

Background: Obesity can lead to the development of type 2 diabetes mellitus (T2DM). However, the predictive power of different obesity anthropometric indices (ObAIs) for T2DM varies with race and geographical area. Therefore, the study aimed to investigate the association between different ObAIs and T2DM and determine the best index for screening T2DM in middle-aged and elderly men and women in Beijing, China. Methods: A cross-sectional study was conducted in Shijingshan district (Beijing, China) from November 2011 to August 2012, involving a total of 14,558 subjects aged ≥40 years. Data on demographic information, lifestyle, history of T2DM, hypertension and dyslipidemia were collected. Body height, body weight, waist circumference (WC), hip circumference, and blood pressure were recorded. The oral glucose tolerance test or a standard meal test and blood lipid test were performed. The relationship between different ObAIs and T2DM was analyzed using multiple logistic regression. Results: After adjustment for age, smoking status, alcohol intake, occupation and education degree, diabetes family history, hypertension and dyslipidemia, body mass index (BMI), WC, waist-to-hip ratio (WHR), and waist-to-height ratio (WHtR) were positively associated with T2DM in both men and women. Compared with the lowest BMI, WC, WHR, and WHtR quartiles, ORs of the highest quartiles were 2.131 (95% CI: 1.465-3.099), 1.752 (95% CI: 1.270-2.417), 1.342 (95% CI: 1.072-1.678), 2.739 (95% CI: 2.293-3.271) in men and 1.837 (95% CI: 1.584-2.130), 3.122 (95% CI: 1.980-4.924), 3.781 (95% CI: 2.855-5.007), 2.379 (85% CI: 2.040-2.775), respectively, in women. The areas under ROC curve of BMI, WC, WHR, and WHtR for men were 0.584 (95% CI: 0.568-0.600), 0.509 (95% CI: 0.492-0.525), 0.501 (95% CI: 0.485-0.518), and 0.642 (95% CI: 0.627-0.658) and 0.619 (95% CI: 0.607-0.632), 0.709 (95% CI: 0.697-0.720), 0.741 (95% CI: 0.730-0.752), and 0.654 (95% CI: 0.642-0.666), respectively, for women. Conclusion: WHtR and WHR have been found to perform better as predictors of T2DM in middle-aged and elderly Chinese men and women, respectively.

Decoding Expression Dynamics of Protein and Transcriptome at the Single-Cell Level in Paired Picoliter Chambers.

Simultaneous analysis of mRNAs and proteins at the single-cell level provides information about the dynamics and correlations of gene and protein expressions in individual cells, enabling a comprehensive study of cellular heterogeneity and expression patterns. Here, we present a platform for about 1000 cellular indexing of mRNAs and membrane proteins, named multi-Paired-seq, with high cell utilization, accurate molecular measurement, and low cost. Based on hydrodynamic differential flow resistance, multi-Paired-seq largely improves cell utilization in the percentage of cells measured in population (>95%). Combined with the pump/valve structure, cell-free antibodies and mRNAs can be removed completely for highly accurate detection (R = 0.96) of protein copies. The picoliter reaction chambers allow high detection sensitivity for both mRNA transcripts and protein copies and low sequencing cost. Using multi-Paired-seq, three clusters of known breast cancer cell types are identified according to multimodal measurements, and the expression correlations between mRNAs and proteins under altered conditions are quantified. Multi-Paired-seq provides multimodal measurements at the single-cell level, which offers a new tool for cell biology, developmental biology, drug discovery, and precision medicine.

Overexpression of AHL9 accelerates leaf senescence in Arabidopsis thaliana.

BACKGROUND: Leaf senescence, the final stage of leaf growth and development, is regulated by numerous internal factors and environmental cues. Ethylene is one of the key senescence related hormones, but the underlying molecular mechanism of ethylene-induced leaf senescence remains poorly understood. RESULTS: In this study, we identified one AT-hook like (AHL) protein, AHL9, as a positive regulator of leaf senescence in Arabidopsis thaliana. Overexpression of AHL9 significantly accelerates age-related leaf senescence and promotes dark-induced leaf chlorosis. The early senescence phenotype observed in AHL9 overexpressing lines is inhibited by the ethylene biosynthesis inhibitor aminooxyacetic acid suggesting the involvement of ethylene in the AHL9-associated senescence. RNA-seq and quantitative reverse transcription PCR (qRT-PCR) data identified numerous senescence-associated genes differentially expressed in leaves of AHL9 overexpressing transgenic plants. CONCLUSIONS: Our investigation demonstrates that AHL9 functions in accelerating the leaf senescence process via ethylene synthesis or signalling.

Generation of a thrombopoietin-deficient thrombocytopenia model in zebrafish.

BACKGROUND: The production of platelets is tightly regulated by thrombopoietin (THPO). Mutations in the THPO gene cause thrombocytopenia. Although mice lacking Thpo present with thrombocytopenia, predicting phenotypes and pathogenicity of novel THPO mutations in mice is limited. Zebrafish can be a powerful tool for fast validation and study of candidate genes of human hematological diseases and have already been used as a model of human thrombocytopenia. OBJECTIVES: We aim to investigate the role of Thpo in zebrafish thrombopoiesis and to establish a Thpo-deficient zebrafish model. The model could be applied for illustrating the clinically discovered human THPO variants of which the clinical significance is not known and to evaluate the effect of THPO receptor agonists (THPO-Ras), as well as a screening platform for new drugs. METHODS: We generated a thpo loss-of-function zebrafish model using CRISPR/Cas9. After disruption of zebrafish thpo, thposzy6 zebrafish presented with a significant reduction of thpo expression and developed thrombocytopenia. Furthermore, we performed in vivo studies with zebrafish with the thposzy6 mutation and found two human clinical point mutations (c.091C > T and c.112C > T) that were responsible for the thrombocytopenia phenotype. In addition, effects of THPO-RAs used as therapeutics against thrombocytopenia were evaluated in the Tg(mpl:eGFP);thposzy6 line. RESULTS AND CONCLUSIONS: Zebrafish with the mutation thposzy6 presented with a significant reduction of thpo expression and developed thrombocytopenia. Thpo loss-of-function zebrafish model can serve as a valuable preclinical model for thrombocytopenia caused by thpo-deficiency, as well as a tool to study human clinical THPO variants and evaluate the effect of THPO-RAs.

Fluorescent Biosensor Based on Hairpin DNA Stabilized Copper Nanoclusters for Chlamydia trachomatis Detection.

Chlamydia trachomatis (C. trachomatis) is a kind of intracellular parasitic microorganism, which can causes many diseases such as trachoma. In this strategy, a specific hairpin DNA with the probe loop as specific regions to recognize C. trachomatis DNA with strong affinity was designed, and its stem consisted of 24 AT base pairs as an effective template for hairpin DNA-CuNCs formation. In the absence of C. trachomatis DNA, the detection system showed strong orange fluorescence emission peaks at 606 nm. In the presence of C. trachomatis DNA, the conformation of DNA probe changed after hybridizing with C. trachomatis DNA. Then, the amount of hairpin DNA-CuNCs was reduced and resulted in low fluorescence emission. C. trachomatis DNA displayed a significant inhibitory effect on the synthesis of fluorescent hairpin DNA-CuNCs due to the competition between C. trachomatis DNA and the specific hairpin DNA. Under the optimal experimental conditions, different concentrations of C. trachomatis were tested and the results showed a good linear relationship in the range of 50 nM to 950 nM. Moreover, the detection limit was 18.5 nM and this detection method possessed good selectivity. Finally, the fluorescent biosensor had been successfully applied to the detection of C. trachomatis target sequence in HeLa cell lysate, providing a new strategy for the detection of C. trachomatis.

Dynamic analysis of a malaria reaction-diffusion model with periodic delays and vector bias.

One of the most important vector-borne disease in humans is malaria, caused by Plasmodium parasite. Seasonal temperature elements have a major effect on the life development of mosquitoes and the development of parasites. In this paper, we establish and analyze a reaction-diffusion model, which includes seasonality, vector-bias, temperature-dependent extrinsic incubation period (EIP) and maturation delay in mosquitoes. In order to get the model threshold dynamics, a threshold parameter, the basic reproduction number $ R_{0} $ is introduced, which is the spectral radius of the next generation operator. Quantitative analysis indicates that when $ R_{0} < 1 $, there is a globally attractive disease-free $ \omega $-periodic solution; disease is uniformly persistent in humans and mosquitoes if $ R_{0} > 1 $. Numerical simulations verify the results of the theoretical analysis and discuss the effects of diffusion and seasonality. We study the relationship between the parameters in the model and $ R_{0} $. More importantly, how to allocate medical resources to reduce the spread of disease is explored through numerical simulations. Last but not least, we discover that when studying malaria transmission, ignoring vector-bias or assuming that the maturity period is not affected by temperature, the risk of disease transmission will be underestimate.

Effects of Salt and Homogenization Processing on the Gastrointestinal Fate of Micro/Nano-Sized Colloidal Particles in Bigeye Tuna (Thunnus obesusis) Head Soup: In vitro Digestion Study.

The effects of condiment (salt) and processing technic (homogenization) on digestion and interfacial properties of micro/nano-sized colloidal particles (MNCPs) in bigeye tuna head soup (BTHS) using simulated gastrointestinal digestion model in vitro were investigated. For MNCPs in BTHS, the triglycerides were wrapped with proteins in the form of a ring. After salting, the average particle size of the MNCPs in salted BTHS (SBTHS) decreased compared with BTHS. However, the partial demulsification phenomenon existed, and part of the protein was encapsulated in some MNCPs. After further homogenization, the average particle size of the MNCPs in homogenous SBTHS (HSBTHS) was further decreased based on SBTHS and the MNCP was rearranged, which changed the original membrane structure. After gastrointestinal digestion, adding salt decreased the release of total fatty acids compared with unsalted. But homogenization processing increased the release of total fatty acids in HSBTHS and there was no significant difference (p ≥ 0.05) between HSBTHS and BTHS. Thus, the decrease in the release of some fatty acids due to adding salt was compensated by homogenization. Therefore, the changes in composition and microstructure of MNCPs induced by salt and homogenization might contribute to the digestion difference of MNCPs.

ß-catenin correlates with the progression of colon cancers and berberine inhibits the proliferation of colon cancer cells by regulating the ß-catenin signaling pathway.

INTRODUCTION: Berberine was one of the active components in Chinese herb and exerted tumor suppressive role in cancer progression, but the exact antitumor mechanism is still not clearly clarified. In the present study, bioinformatics analysis was performed on COAD patients from TCGA, HPA database, UALCAN and GEPIA 2 platform. We also explored the role of berberine on progression of human colon cancers in vitro and in vivo and clarified weather the antitumor effects of berberine was mediated by Wnt/beta-catenin pathway. METHODS: Cell viability was determined by MTT assay. The protein levels were tested by western blotting and the distribution of ß-catenin was observed by confocal microscope. RESULTS: The results showed the levels of CTNNB1 mRNA was increased in colon cancer patients than normal controls. The diagnostic value of CTNNB1 was AUC = 0.882 (CI:0.854-0.911) with sensitivity of 1.000 and specificity of 0.777. The promoter methylation level of CTNNB1 in COAD patients was significantly decreased. Moreover, univariate analysis and multivariate analysis results showed the expression of CTNNB1 in COAD patients was associated with T stage (p = 0.010), pathological stage (p = 0.025) and perineural invasion (p = 0.025). Furthermore, the in vitro assay results showed ß-catenin signaling was highly activated in human colon cancer cells and berberine inhibited the cell viability of colon cancer cells in vitro and in vivo in a dose-and time-dependent manner. Moreover, berberine induced the translocation of ß-catenin to cytoplasm from nucleus. CONCLUSION: The levels of CTNNB1 mRNA was increased in colon cancer patients than normal controls. Berberine inhibited the proliferation of colon cancer cells by regulating the beta-catenin signaling pathway.

Colonization and Development of the Fecal Microflora of South China Tiger Cubs (Panthera tigris amoyensis) by Sequencing of the 16S rRNA Gene.

Postnatal colonization and development of the gut microbiota is linked to health and growth. A comprehensive understanding of the postnatal compositional changes and development of the microbial community is helpful to understand the gut health and improve the survival rate of South China tiger cubs (Panthera tigris amoyensis). Fecal samples from three tiger cubs were collected on the day of birth in 2018 (June 17-21 [G0], July 18 [G1], July 31 [G2], and August 7 [G3]). The 16S rRNA genes of the fecal microflora were sequenced. Results showed that 38 phyla, 58 classes, 134 orders, 272 families, and 636 genera of bacteria from 3,059 operational taxonomic units were identified from 12 fecal samples. The diversity and abundance of species of group G0 were significantly higher (p < 0.05 or 0.01) than those of groups G2 and G3. The predominant phylum was Proteobacteria in groups G0 and G1 (38.85% and 48%, respectively) and Firmicutes in groups G2 and G3 (71.42% and 75.29%, respectively). At the phylum level, the abundance of Deinococcus-Thermus was significantly decreased in groups G1, G2, and G3 as compared to group G0 (p < 0.05), while that of Firmicutes was significantly increased in groups G2 and G3 (p < 0.05). At the genus level, the abundance of Faecalibacterium, Ralstonia, and unidentified Rickettsiales was significantly decreased in groups G1, G2, and G3 as compared with group G0 (p < 0.05), while that of Pseudomonas was significantly decreased in groups G2 and G3 (p < 0.05). The composition and structure of fecal microbiota of South China tiger cubs changed after birth.

[Sequence analysis for a case with Para-Bombay AB blood type].

OBJECTIVE: To explore the molecular basis for a rare case with Para-Bombay AB blood type. METHODS: Serological method was used to determine the blood type of the proband. Exons 6 and 7 of the ABO gene and the coding regions of FUT1 and FUT2 genes were analyzed by direct sequencing. RESULTS: Serological results showed that the proband was a Para-Bombay AB subtype. His genotype was determined as ABO*A1.02/B.01. The proband was also found to harbor c.551-552delAG and c.881-882delTT of the FUT1 gene. For his four children, there were three type B and one type A, though the expression of the H type was normal. CONCLUSION: The double deletions in the coding region of the FUT1 gene probably underlay the Para-Bombay blood type in the proband. Carrier of single-strand deletions may have a normal ABO phenotype.

Safety of PD-1/PD-L1 Inhibitors Combined With Palliative Radiotherapy and Anti-Angiogenic Therapy in Advanced Hepatocellular Carcinoma.

BACKGROUND: Previous studies have explored cancer immunotherapy with radiotherapy or anti-angiogenic therapy, but no trials have reported a triple therapy approach. This study aimed to investigate safety and clinical outcome of PD-1/PD-L1 inhibitors combined with palliative radiotherapy and targeted angiogenesis therapy in hepatocellular carcinoma (HCC) of Barcelona Clinic Liver Cancer (BCLC) stage C. METHODS: Consecutive patients (n=16) treated with PD-1/PD-L1 inhibitors combined with radiotherapy and anti-angiogenic therapy in a bi-institutional cohort between July 2017 and December 2020 were retrospectively included. Radiotherapy was conducted within 14 days of the first administration of immunotherapy. The primary endpoint was treatment-related adverse event (TRAE). RESULTS: The median follow-up was 383 days. Fifteen patients (93.8%) experienced at least 1 TRAE. The most common TRAEs of any grade were rash (25%), diarrhea (25%), aspartate aminotransferase increase (18.8%), alanine transaminase increase (18.8%), decreased appetite (18.8%), and fatigue (18.8%). Grade 3/4 TRAEs occurred in 4 patients (25%) and finally led to treatment interruption. No patient death was attributed to treatment. No specific events were responsible for the addition of radiotherapy. Six patients showed partial response, 7 showed stable disease, and 2 showed progressive disease. The objective response rate and disease control rate were 40.0% (95% CI 16.3%-67.7%) and 86.7% (95% CI 59.5%-98.3%), respectively. Moreover, the median progression-free survival was 140 days. Patients had a median overall survival of 637 days, and the estimated rates of survival at 6 and 12 months were 92.3% and 75.5%, respectively. CONCLUSION: PD-1/PD-L1 inhibitors combined with palliative radiotherapy and anti-angiogenic therapy appear to be safe, with no unexpected adverse events. Additional studies exploring the clinical benefit are warranted.