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Introduction: Non-alcoholic fatty liver disease (NAFLD) is gradually becoming one of the most common chronic liver diseases worldwide. Compared to chemical drugs, traditional Chinese medicine (TCM) has significant advantages in the treatment of NAFLD owing to its long history, remarkable efficacy, and low toxicity. Methods: The scientific databases, including PubMed, Web of Science, and China National Knowledge Infrastructure database (CNKI) were searched using the keywords of NAFLD, TCM, lipid metabolism, high-fat diet, bupleurum decoction, Yinchenhao decoction, Shugan Jiangzhi granules, Shugan Jianpi Jiangzhuo decoction, GanShuang granules, Gegen Qinlian decoction, bupleurum, Yinchen, Jiaogulan, Huangqin, Gegen, Baizhu, flavonoids, baicalein, luteolin, kaempferol, quercetin, apigenin, isorhamnetin, hesperidin, icariin, rutin, soy isoflavones, puerarin, phlorizin, total flavonoids from hawthorn leaves, and total flavonoids from buckwheat flowers and leaves to overview the pathogenesis and treatment of NAFLD. Results: This review explained the causes of NAFLD, including factors like lipotoxicity, insulin resistance, ER stress, mitochondrial dysfunction, oxidative stress, gut dysbiosis, and inflammation. Furthermore, the review provided a thorough overview of the effects and mechanisms of TCM in the treatment of NAFLD. This included an analysis of six TCM formulas, six single herbs, and 14 flavonoids derived from TCM. Discussion: TCM has proven to be highly effective in the treatment of NAFLD, with numerous Chinese medicine formulas and individual herbs being commonly used in clinical practice. The active components in these TCM, flavonoids, have demonstrated significant biological effects in addressing NAFLD. Conclusion: TCM has the potential to greatly enhance the treatment of NAFLD with minimal adverse reactions. Given these results, we are confident that TCM holds promising opportunities for the management of NAFLD and can offer valuable perspectives for future research.

Summary: Integrins play critical roles in connecting the extracellular matrix and actin. While the upregulation of integrins is thought to promote cancer stemness and metastasis, the mechanisms underlying their upregulation in cancer stem cells (CSCs) remain poorly understood. Herein, we show that USP22 is essential in maintaining breast cancer cell stemness by promoting the transcription of integrin β1 (ITGB1). Both genetic and pharmacological inhibition of USP22 largely impaired breast CSCs self-renewal and prevented their metastasis. Reconstitution of integrin β1 partially rescued USP22-null breast cancer metastasis. USP22 functions as a bona fide deubiquitinase to protect the proteasomal degradation of the forkhead box M1 (FoxM1), a transcription factor for tumoral ITGB1 gene transcription. Immunohistochemistry staining detected a positive correlation among USP22, FoxM1, and integrin β1 in human breast cancers. Collectively, our study identifies the USP22-FoxM1-integrin β1 signaling axis as critical for cancer stemness and offers a potential target for antitumor therapy.

Background: Serum prostate-specific antigen (PSA) is a primary metric for diagnosis and prognosis of prostate cancer (PCa). Exposure to heavy metals, such as lead, cadmium, mercury, and zinc can impact PSA levels in PCa patients. However, it is unclear whether this effect also occurs in men without PCa, which may lead to the overdiagnosis of PCa. Method: Data on a total of 5089 American men who had never been diagnosed with PCa were obtained from the National Health and Nutrition Examination Survey performed from 2003–2010. The relationship between serum PSA levels (dependent variable) and concentrations of lead (μmol/L), cadmium (nmol/L), and mercury (μmol/L) were investigated with dietary zinc intake being used as a potential modifier or covariate in a weighted linear regression model and a generalized additive model. A series of bootstrapping analyses were performed to evaluate sensitivity and specificity using these models. Results: Regression analyses suggested that, in general, lead, cadmium, or mercury did not show an association with PSA levels, which was consistent with the results of the bootstrapping analyses. However, in a subgroup of participants with a high level of dietary zinc intake (≥14.12 mg/day), a significant positive association between cadmium and serum PSA was identified (1.06, 95% CI, P = 0.0268, P for interaction=0.0249). Conclusions: With high-level zinc intake, serum PSA levels may rise in PCa-free men as the exposure to cadmium increases, leading to a potential risk of an overdiagnosis of PCa and unnecessary treatment. Therefore, environmental variables should be factored in the current diagnostic model for PCa that is solely based on PSA measurements. Different criteria for PSA screening are necessary based on geographical variables. Further investigations are needed to uncover the biological and biochemical relationship between zinc, cadmium, and serum PSA levels to more precisely diagnose PCa.

Abstract Background Gene knockout and knock-in have been widely performed in large farm animals based on genome editing systems. However, many types of precise gene editing, including targeted deletion, gene tagging, and large gene fragment replacement, remain a challenge in large farm animals. Results Here, we established versatile self-excising gene-targeting technology in combination with programmable nucleases (SEGCPN) to efficiently generate various types of precise gene editing in bovine. First, we used this versatile method to successfully generate bovine embryos with point mutations and 11-bp deletions at the MSTN locus. Second, we successfully generated bulls with EGFP labeling at the SRY locus. Finally, we successfully generated humanized cows in which the endogenous 18-kb α-casein gene was replaced with a 2.6-kb human α-lactalbumin gene. Conclusions In summary, our new SEGCPN method offers unlimited possibilities for various types of precise gene editing in large animals for application both in agriculture and disease models.

Abstract Background Microbial residues are significant contributors to stable soil organic carbon (SOC). Soil aggregates effectively protect microbial residues against decomposition; thus, microbial residue accumulation and distribution among soil aggregates determine long-term SOC stability. However, how tree species influence accumulation and distribution of soil microbial residues remains largely unknown, hindering the chances to develop policies for SOC management. Here, we investigated microbial residue accumulation and distribution in soil aggregates under four subtropical tree species (Cunninghamia lanceolata, Pinus massoniana, Michelia macclurei, and Schima superba) after 29 years of afforestation. Results Accumulation of microbial residues in the 0–10 cm soil layer was 13.8–26.7% higher under S. superba than that under the other tree species. A structural equation model revealed that tree species affected the accumulation of microbial residues directly by altering fungal biomass. Additionally, tree species significantly affected microbial residue distribution and contribution to SOC in the top 20 cm soil. In particular, microbial residue distribution was 17.2–33.4% lower in large macro-aggregates (LMA) but 60.1–140.7% higher in micro-aggregates (MA) under S. superba than that under the other species in the 0–10 cm soil layer, and 14.3–19.0% lower in LMA but 43–52.1% higher in MA under S. superba than that under C. lanceolata and M. macclurei in the 10–20 cm soil layer. Moreover, the contribution of microbial residues to SOC was 44.4–47.5% higher under S. superba than under the other tree species. These findings suggest a higher stability of microbial residues under S. superba than that under the other studied tree species. Conclusions Our results demonstrate that tree species influence long-term microbial persistence in forest soils by affecting accumulation and stabilization of microbial residues.

Abstract. Integrins are a family of transmembrane receptors that connect the extracellular matrix and actin skeleton, which mediate cell adhesion, migration, signal transduction, and gene transcription. As a bi-directional signaling molecule, integrins can modulate many aspects of tumorigenesis, including tumor growth, invasion, angiogenesis, metastasis, and therapeutic resistance. Therefore, integrins have a great potential as antitumor therapeutic targets. In this review, we summarize the recent reports of integrins in human hepatocellular carcinoma (HCC), focusing on the abnormal expression, activation, and signaling of integrins in cancer cells as well as their roles in other cells in the tumor microenvironment. We also discuss the regulation and functions of integrins in hepatitis B virus-related HCC. Finally, we update the clinical and preclinical studies of integrin-related drugs in the treatment of HCC.

Abstract Background Currently, there are relatively few studies on the effects of changes in oestrogen and androgen levels on prostatic microvessel density (MVD). This article aimed to study the changes in prostatic MVD in Sprague-Dawley (SD) rats after castration under the effect of oestrogen/androgen at different concentrations. Methods Male SD rats aged 3–4 months were randomly divided into a control group, a castration group, and groups with different concentrations of oestrogen/androgen treatment after castration. Dihydrotestosterone (DHT) and oestradiol (E) were administered daily by subcutaneous injection for one month. All the rats were killed by cervical dislocation after one month, and the serum DHT and E concentrations of the rats in each group were measured by ELISA. Prostate tissue specimens were immunohistochemically stained with monoclonal antibodies against CD34 and factor VIII for MVD. Results Compared with the control group, the MVD decreased significantly in the castration group (P

Background: The most common disease caused by biallelic AFG3L2 mutations is spastic ataxia type 5 (SPAX5). Identification of complex phenotypes resulting from biallelic AFG3L2 mutations has been increasing in recent years. Methods: A retrospective analysis was performed on a child with microcephaly and recurrent seizures. The child underwent physical and neurological examinations, laboratory tests, electroencephalography (EEG), and brain magnetic resonance imaging (MRI). Trio-whole-exome sequencing (trio-WES) was performed to identify possible causative mutations. Results: We described a child who exhibited early-onset and intractable epilepsy, developmental regression, microcephaly, and premature death. Neuroimaging revealed global cerebral atrophy (GCA) involving the cerebrum, cerebellum, corpus callosum, brainstem, cerebellar vermis, and basal ganglia. On trio-WES, two novel compound heterozygous mutations, c.1834G > T (p.E612*) and c.2176-6T > A in the AFG3L2 gene, were identified in this patient. Conclusions: Our findings have expanded the mutation spectrum of the AFG3L2 gene and identified a severe neurodegenerative phenotype of global cerebral atrophy caused by biallelic AFG3L2 mutations.

Temperature and precipitation patterns during the Last Glacial Maximum are better predictors of global-scale variations in the temperature sensitivity of soil respiration in response to nitrogen enrichment than contemporary climate, according to a global meta-analysis and structural equation model.

Abstract Background The correlation between modified bladder outlet obstruction index (MBOOI) and surgical efficacy still remains unknown. The purpose of the study was to investigate the clinical value of the MBOOI and its use in predicting surgical efficacy in men receiving transurethral resection of the prostate (TURP). Methods A total of 403 patients with benign prostate hyperplasia (BPH) were included in this study. The International Prostate Symptom Score (IPSS), quality of life (QoL) index, transrectal ultrasonography, and pressure flow study were conducted for all patients. The bladder outlet obstruction index (BOOI) (PdetQmax–2Qmax) and MBOOI (Pves–2Qmax) were calculated. All patients underwent TURP, and surgical efficacy was accessed by the improvements in IPSS, QoL, and Qmax 6 months after surgery. The association between surgical efficacy and baseline factors was statistically analyzed. Results A comparison of effective and ineffective groups based on the overall efficacy showed that significant differences were observed in PSA, Pves, PdetQmax, Pabd, BOOI, MBOOI, TZV, TZI, IPSS-t, IPSS-v, IPSS-s, Qmax, and PVR at baseline (p

Abstract Background Inputs of above- and belowground litter into forest soils are changing at an unprecedented rate due to continuing human disturbances and climate change. Microorganisms drive the soil carbon (C) cycle, but the roles of above- and belowground litter in regulating the soil microbial community have not been evaluated at a global scale. Methods Here, we conducted a meta-analysis based on 68 aboveground litter removal and root exclusion studies across forest ecosystems to quantify the roles of above- and belowground litter on soil microbial community and compare their relative importance. Results Aboveground litter removal significantly declined soil microbial biomass by 4.9% but root exclusion inhibited it stronger, up to 11.7%. Moreover, the aboveground litter removal significantly raised fungi by 10.1% without altering bacteria, leading to a 46.7% increase in the fungi-to-bacteria (F/B) ratio. Differently, root exclusion significantly decreased the fungi by 26.2% but increased the bacteria by 5.7%, causing a 13.3% decrease in the F/B ratio. Specifically, root exclusion significantly inhibited arbuscular mycorrhizal fungi, ectomycorrhizal fungi, and actinomycetes by 22.9%, 43.8%, and 7.9%, respectively. The negative effects of aboveground litter removal on microbial biomass increased with mean annual temperature and precipitation, whereas that of root exclusion on microbial biomass did not change with climatic factors but amplified with treatment duration. More importantly, greater effects of root exclusion on microbial biomass than aboveground litter removal were consistent across diverse forest biomes (expect boreal forests) and durations. Conclusions These data provide a global evidence that root litter inputs exert a larger control on microbial biomass than aboveground litter inputs in forest ecosystems. Our study also highlights that changes in above- and belowground litter inputs could alter soil C stability differently by shifting the microbial community structure in the opposite direction. These findings are useful for predicting microbe-mediated C processes in response to changes in forest management or climate.

Cancer vaccines have gradually attracted attention for their tremendous preclinical and clinical performance. With the development of next-generation sequencing technologies and related algorithms, pipelines based on sequencing and machine learning methods have become mainstream in cancer antigen prediction; of particular focus are neoantigens, mutation peptides that only exist in tumor cells that lack central tolerance and have fewer side effects. The rapid prediction and filtering of neoantigen peptides are crucial to the development of neoantigen-based cancer vaccines. However, due to the lack of verified neoantigen datasets and insufficient research on the properties of neoantigens, neoantigen prediction algorithms still need to be improved. Here, we recruited verified cancer antigen peptides and collected as much relevant peptide information as possible. Then, we discussed the role of each dataset for algorithm improvement in cancer antigen research, especially neoantigen prediction. A platform, Cancer Antigens Database (CAD, http://cad.bio-it.cn/), was designed to facilitate users to perform a complete exploration of cancer antigens online.

Reactivation of fetal hemoglobin (HbF, α2γ2) alleviates clinical symptoms in patients with β-thalassemia and sickle cell disease, although the regulatory mechanisms of γ-globin expression have not yet been fully elucidated. Recent studies found that interfering with the expression of the membrane protein ANTXR1 gene upregulated γ-globin levels. However, the exact mechanism by which ANTXR1 regulates γ-globin levels remains unclear. Our study showed that overexpression and knockdown of ANTXR1 in K562, cord blood CD34+, and HUDEP-2 cells decreased and increased γ-globin expression, respectively. ANTXR1 regulates the reactivation of fetal hemoglobin (HbF, α2γ2) in K562, cord blood CD34+, and adult peripheral blood CD34+ cells through interaction with LRP6 to promote the nuclear entry of β-catenin and activate the Wnt/β-catenin signaling pathway. The overexpression or knockdown of ANTXR1 on γ-globin and Wnt/β-catenin signaling in K562 cells was reversed by the inhibitor XAV939 and the activator LiCl, respectively, where XAV939 inhibits the transcription of β-catenin in the Wnt pathway, but LiCl inhibits GSK3-β. We also showed that the binding ability of the rank4 site in the transcriptional regulatory region of the SOX6 gene to c-Jun was significantly increased after overexpression of ANTXR1 in K562 cells. SOX6 protein expression was increased significantly after overexpression of the c-Jun gene, indicating that the transcription factor c-Jun initiated the transcription of SOX6, thereby silencing γ-globin. Our findings may provide a new intervention target for the treatment of β-hemoglobinopathies.

The replenishment and priming effect (PE) are two decisive processes that determine the carbon (C) sequestration potential of biochar. However, how increased nitrogen (N) availability affect these two processes and the consequent net C balance remains poorly understood. By collecting soils from three forest ecosystems (deciduous broad-leaf forest (DBF), evergreen coniferous forest (ECF), and evergreen broad-leaf forest (EBF)), we conducted a 365-day incubation experiment by adding 13C-labelled biochar plus five rates of inorganic N (0 to 15% N of soil total N). The -results showed that N addition significantly stimulated the early period (0–48 days) but did not affect the late period (49–365 days) of biochar decomposition. The effect of N addition on PE varied largely with the forest type and decomposition period; N addition significantly enhanced the negative PE -in both periods in DBF and at the late period in EBF, whereas it stimulated positive PE in the early period in EBF and ECF. At the end of incubation, the addition of biochar caused net C accumulation across all treatments due to the huge proportion of biochar (98.1%–98.9% of added biochar) retained in soils and the negative or neutral cumulative PE (−11.25–0.35 g C kg−1 SOC), and the magnitude of net C balance increased linearly with the N addition rate in DBF and EBF. Collectively, the results of this study indicate that biochar input can contribute to soil C sequestration and that N addition can enhance the C sequestration potential of biochar.

Forest soil respiration is an important part of the terrestrial ecosystem carbon (C) cycle and controlled by multiple factors, especially atmospheric nitrogen (N) deposition and environmental change. However, it remains unclear how soil respiration responds to N deposition in forests along an urban–rural gradient, as most studies have only been conducted in single environmental areas, especially in natural forests. Here, in order to explore how N deposition influences soil respiration, a field experiment with different levels of N deposition (50 and 100 kg N·ha−1·yr−1) was conducted in forests along an urban–rural gradient of Hefei (31°52′N, 117°17′ E). Soil temperature (T), soil water content (SW), soil physiochemical properties, and soil microbes were simultaneously measured to investigate the underlying mechanisms of N deposition effects on soil respiration. The results showed that the soil respiration and cumulative emission flux of CO2 (RCUM) were increased by N deposition in all forests. Nitrogen addition treatments significantly enhanced the mean RCUM by approximately 6.04%, 11.02%, and 3.62%, respectively, in urban, suburban, and rural forests. The positive effects of N deposition on soil respiration had different temporal variations in different forests, and the strongest positive effects occurred with the shortest time was found in the urban forest. Temperature sensitivities of soil (Q10) were significantly different in the three forests, showing higher Q10 values in forests more near to the urban. In all forests, the total interpretation rates of climate factors, soil physicochemical properties and soil microbes to soil respiration were over 80%. Climate factors were the primary factor for explaining the variation of soil respiration under N deposition. The interpretations of the climate factors, soil physiochemical properties, and soil microbes to soil respiration were changed by N deposition, and the combined effects of these factors were much higher than those of the individual factors. Our findings provide novel insights for studying the response of soil respiration to N deposition under different environmental factors.

Abstract Background The development of ureteral calculi after Yang-Monti ileal ureter reconstruction has not been reported. This study was performed to explore the safety and effectiveness of ureteroscopy combined with laser lithotripsy in the treatment of ipsilateral lower ureteral calculi and lower calyceal calculi after Yang-Monti ileal ureter reconstruction. Case presentation A 48-year-old man was admitted to our hospital with ipsilateral distal ureteral calculi and ipsilateral lower calyceal calculi. One year prior to this admission, the patient had undergone Yang-Monti ileal ureter reconstruction due to long-segment ureteral stenosis. After conservative treatment failed, we used a rigid ureteroscope with a holmium laser to break up the distal ureteral calculi, and successfully removed the renal calculi with a digital flexible ureteroscope and basket extractor. Conclusion The successful outcome of the present case suggests that ureteroscopy combined with laser lithotripsy is a valuable option for the management of urinary calculi following Yang-Monti ileal ureter reconstruction.

Soil heterotrophic respiration (Rh) is an important pathway of carbon (C) dioxide release from terrestrial soils to the atmosphere. It is often measured using sieved soil in a laboratory, but the uncertainty of how it is influenced by soil sieving persists, which limits the accuracy of predicting soil organic C dynamics in C models. To address how soil sieving during laboratory incubation affects Rh and its response to increased carbon availability, we investigated Rh in sieved and intact soil cores and its response to 13C-glucose addition. This was conducted through a 27-day laboratory incubation in four forests, including two ectomycorrhizal-dominated (ECM) forests and two arbuscular mycorrhizal-dominated forests. The significant influence of soil sieving on Rh in all forests was not observed during incubation when glucose was not added. After adding glucose, the Rh in the sieved soils on the 5th day of incubation was averaged 27.2% lower than that in intact soils in ECM forests. On the 27th day it was 22.1% lower in the Pinus massoniana forest, but 78.0% higher in the Castanea mollissima forest. Strong relationships were detected between Rh in sieved and intact soils (r2 = 0.888), and in soils both with and without the addition of glucose (r2 = 0.827). The measured soil variables explained 74.7% and 49.7% of the variation in Rh on the 5th and 27th day of incubation, and the role of soil nutrients and microbial PLFA groups in regulating Rh varied temporally. Our findings suggest that plant mycorrhizal types influenced the role of increased C availability to microbes in regulating the response of Rh to sieving in forest ecosystems.

Soil microbial residues contribute to the majority of stable soil organic carbon (SOC) pools, and their distribution among aggregate fractions determines long-term soil carbon (C) stability and, consequently, soil productivity. However, how microbial residue accumulation and distribution respond to stand age remains unexplored. To fill this knowledge gap, we investigated microbial residues in bulk soil and soil aggregate fractions under a chronosequence of Chinese fir (Cunninghamia lanceolata [Lamb.] Hook) plantations with stands aged 3, 17, 27, and 36 years. The results showed that microbial residues in topsoil did not change across the different stand ages, but the residues in the subsoil increased from 3 to 17 years of age and then remained constant. Moreover, microbial residue distribution in microaggregates decreased with stand age, and the residue distribution in small macroaggregates was lower at age 17 years than at other stand ages. The effect of stand age on microbial residue distribution was due to the fact of their effect on aggregate distribution but not microbial residue concentrations in aggregate fractions. Collectively, our results indicate that microbial residue stability decreased with stand age, which has significant implications for the management of SOC in subtropical plantations.

Abstract Increasing fire risk and atmospheric nitrogen (N) deposition have the potential to alter plant community structure and composition, with consequent impacts on biodiversity and ecosystem functioning. This study was conducted to examine short‐term responses of understory plant community to burning and N addition in a coniferous‐broadleaved mixed forest of the subtropical‐temperate transition zone in Central China. The experiment used a pair‐nested design, with four treatments (control, burning, N addition, and burning plus N addition) and five replicates. Species richness, cover, and density of woody and herbaceous plants were monitored for 3 years after a low‐severity fire in the spring of 2014. Burning, but not N addition, significantly stimulated the cover (+15.2%, absolute change) and density (+62.8%) of woody species as well as herb richness (+1.2 species/m2, absolute change), cover (+25.5%, absolute change), and density (+602.4%) across the seven sampling dates from June 2014 to October 2016. Light availability, soil temperature, and prefire community composition could be primarily responsible for the understory community recovery after the low‐severity fire. The observations suggest that light availability and soil temperature are more important than nutrients in structuring understory plant community in the mixed forest of the subtropical‐temperate transition zone in Central China. Legacy woody and herb species dominated the understory vegetation over the 3 years after fire, indicating strong resistance and resilience of forest understory plant community and biodiversity to abrupt environmental perturbation.

Fuzheng Huayu (FZHY) capsule is a traditional Chinese medicine composed of six Chinese medicinal herbs Tian et al. [1] and approved by China food and drug administration for liver fibrosis treatment [2,3] Liu et al., 2009 and Liu et al., 2005. CGA formula consisting of Cordyeps sinensis polysaccharide (CS-PS), gypenosides (G), and amygdalin (A), are derived from FZHY formula. It is necessary to identify the chemical profile of FZHY and CGA formula to describe the mechanisms and the corresponding components of anti-fibrosis. It is showed that FZHY contains adenosine (5.21 mg/g), amygdalin (5.31 mg/g), salvianolic acid b (18.22 mg/g) and deoxyschizandrin (2.62 mg/g), respectively. CS-PS contained 60.5 ± 2.2% total carbohydrate, including 14.17% arabinose, 25.35% glucose and 60.48% galactose. Gypenosides contain 10.34% gypenosides XLIX and 16.58% gypenosides A. These data provide the primary chemical profile of FZHY and CGA formula and an example for components analysis of traditional Chinese medicine. Keywords: High-performance liquid chromatography, Traditional Chinese medicine, Fuzheng huayu capsule, Cordyeps sinensis polysaccharide, Gypenosides

Objective: Inspired by the traditionally clinical application of herb pair Zhimu-Huangbo to treat diabetes, a combination of plant ingredients, timosaponin B2 (TB-2) and berberine (BBR), was evaluated for their anti-diabetic efficacy and cooperative mechanisms.Methods: The efficacy and pharmacokinetics of orally administered TB-2 (33.3 mg/kg/day), BBR (66.7 mg/kg/day), and TB-2+BBR (100 mg/kg/day) were evaluated in spontaneously non-obese diabetic Goto-Kakizaki (GK) rats, and metformin (200 mg/kg/day) was used as a positive control. The comparative exposure of the parent drugs, timosaponin A3 (TB-2 metabolite), and M1–M5 (BBR metabolites) was quantified in the portal vein plasma (before hepatic disposition), liver, and systemic plasma (after hepatic disposition) of normal rats on single and combination treatments. Cooperative mechanism of TB-2 and BBR on intestinal absorption and hepatic metabolism was investigated in Caco-2 cells and primary hepatocytes, respectively.Results: After a 6-week experiment, non-fasting and fasting blood glucose levels and oral glucose tolerance test results showed that TB-2+BBR treatments (100 mg/kg/day) displayed significantly anti-diabetic efficacy in GK rats, comparable to that on metformin treatments. However, no significant improvement was observed on TB-2 or BBR treatments alone. Compared to single treatments, combination treatments led to the increased circulating levels of BBR by 107% in GK rats. In normal rats, the hepatic exposure of BBR, timosaponin A3, and M1–M5 was several hundred folds higher than their circulating levels. Co-administration also improved the levels in the plasma and liver by 41–114% for BBR, 141–230% for TB-2, and 12–282% for M1–M5. In vitro, the interaction between TB-2 and BBR was mediated by intestinal absorption, rather than hepatic metabolism.Conclusion: Combining TB-2 and BBR enhanced the anti-diabetic efficacy by increasing the in vivo variety of effective substances, including the parent compounds and active metabolites, and improving the levels of those substances through intestinal absorption. This study is a new attempt to assess the effects of combined plant ingredients on diabetes by scientifically utilizing clinical experience of an herb pair.

ABSTRACT Objective To evaluate the clinical efficiency of alpha1-adrenergic antagonists on stentless ureteroscopic lithotripsy treating uncomplicated lower ureteral stones. Materials and Methods From January 2007 to January 2013, 84 patients who have uncomplicated lower ureteral stones treated by ureteroscopic intracorporeal lithotripsy with the holmium laser were analyzed. The patients were divided into two groups, group A (44 patients received indwelled double-J stents) and group B (40 patients were treated by alpha1-adrenergic antagonists without stents). All cases of group B were treated with alpha1 blocker for 1 week. Results The mean operative time of group A was significantly longer than group B. The incidences of hematuria, flank/abdominal pain, frequency/urgency after surgery were statistically different between both groups. The stone-free rate of each group was 100%. Conclusions The effect of alpha1-adrenergic antagonists is more significant than indwelling stent after ureteroscopic lithotripsy in treating uncomplicated lower ureteral stones.

Cell-penetrating peptides (CPPs) have been increasingly used to deliver various molecules, both in vitro and in vivo. However, there are no reports of CPPs being used in porcine fetal fibroblasts (PFFs). The increased use of transgenic pigs for basic research and biomedical applications depends on the availability of technologies for efficient genetic-modification of PFFs. Here, we report that three CPPs (CPP5, TAT, and R9) can efficiently deliver active Cre recombinase protein into PFFs via an energy-dependent endocytosis pathway. The three CPP-Cre proteins can enter PFFs and subsequently perform recombination with different efficiencies. The recombination efficacy of CPP5-Cre was found to be nearly 90%. The rate-limiting step for CPP-Cre-mediated recombination was the step of endosome escape. HA2 and chloroquine were found to improve the recombination efficiency of TAT-Cre. Furthermore, we successfully obtained marker-free transgenic pigs using TAT-Cre and CPP5-Cre. We provide a framework for the development of CPP-based farm animal transgenic technologies that would be beneficial to agriculture and biomedicine.

Induced pluripotent stem cells (iPSCs) generated from somatic cells by ectopic expression of reprogramming factors, e.g., POU5F1 (OCT4), KLF4, and SOX2, have great potential for regenerative medicine. However, before they can be used in a clinical setting, the mechanism of reprogramming needs to be better understood. Here, by engineering reprogramming factors to a destabilizing protein domain, we achieved inducible generation of mouse and pig iPSCs. Stability of the fusion protein was precisely regulated by the addition of the cell-permeable small molecule trimethoprim (TMP) in a dose-dependent manner. With these tools, we found that during the early and middle stages of reprogramming, exogenous OCT4 or KLF4 could be omitted, whereas exogenous SOX2 expression at early and middle stages was required for successful reprogramming. Our TMP reprogramming system is useful for defining the stoichiometry and temporal requirements of transcription factors for reprogramming.

Cell penetrating peptides (CPPs) are short peptides that can pass through cell membranes. CPPs can facilitate the cellular entry of proteins, macromolecules, nanoparticles and drugs. RVG peptide (RVG hereinafter) is a 29-amino-acid CPP derived from a rabies virus glycoprotein that can cross the blood-brain barrier (BBB) and enter brain cells. However, whether RVG can be used for genome editing in the brain has not been reported. In this work, we combined RVG with Cre recombinase for bacterial expression. The purified RVG-Cre protein cut plasmids in vitro and traversed cell membranes in cultured Neuro2a cells. By tail vein-injecting RVG-Cre into Cre reporter mouse lines mTmG and Rosa26lacZ, we demonstrated that RVG-Cre could target brain cells and achieve targeted somatic genome editing in adult mice. This direct delivery of the gene-editing enzyme protein into mouse brains with RVG is much safer than plasmid- or viral-based methods, holding promise for further applications in the treatment of various brain diseases.