The Chinese medicine Fufang Zhenzhu Tiaozhi capsule protects against renal injury and inflammation in mice with diabetic kidney disease.

ETHNOPHARMACOLOGICAL RELEVANCE: Fufang Zhenzhu Tiaozhi capsule (FTZ) is a patented preparation of Chinese herbal medicine that has been used to treat hyperlipidemia, nonalcoholic fatty liver disease, atherosclerosis, and other glucolipid metabolic diseases (GLMDs) in the clinic for almost 10 years. However, how FTZ reduces albuminuria and attenuates diabetic kidney disease (DKD) progression is unknown. AIM OF THE STUDY: To clarify the effects of FTZ on DKD mice model and to explore the underlying mechanisms. MATERIALS AND METHODS: We used streptozotocin (STZ) (40 mg/kg/d, i.p. for 5 days, consecutively) combined with a high-fat diet (HFD) to induce a DKD mouse model, followed by FTZ (1, 2 g/kg/d, i.g.) treatment for 12 weeks. Losartan (30 mg/kg/d, i.g.) was used as a positive control. Measurements of 24 h proteinuria, serum creatinine (SCr), fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), and low density lipoprotein cholesterol (LDL-C) levels and expression levels of fibronectin (FN), collagen IV, inflammatory cytokines, inflammatory cells, interleukin-17A (IL-17A) and the nuclear transcription factor-κB (NF-κB) signaling pathway in the kidney were examined. RESULTS: FTZ effectively decreased 24 h proteinuria, Scr, FBG, TC, TG, and LDL-C levels, inhibited mesangial cell expansion, reduced FN and collagen IV accumulation, and F4/80+ macrophage cell infiltration and Ly-6G+ neutrophil infiltration in glomerulus and tubulointerstitium. Furthermore, IL-17A production and the NF-κB signaling pathway were also downregulated after the administration of FTZ. CONCLUSION: FTZ might attenuate DKD progression, and inhibited kidney inflammation and fibrosis by inhibiting the expression of RORγT and IL-17A in vivo, offering novel insights for the clinical application of FTZ.

The effect of atrazine on intestinal histology, microbial community and short chain fatty acids in Pelophylax nigromaculatus tadpoles.

The intestine is the main organ for nutrient absorption in amphibians. It is sensitive to atrazine, which is a herbicide widely used in agricultural areas. At present, there is a lack of systematic research on the effect of atrazine on the amphibian intestine. In this study, we evaluated the effects of atrazine exposure (0, 50 µg/L, 100 µg/L, and 500 µg/L) for 20 days on intestinal histology, microbiota and short chain fatty acids in Pelophylax nigromaculatus tadpoles. Our research showed that 500 µg/L atrazine exposure significantly decreased the height of microvilli and epithelial cells, and altered the composition and diversity of intestinal microbiota in P. nigromaculatus tadpoles compared to the control. At the phylum level, the abundance of Bacteroidetes and Fusobacteria increased significantly, while that of Verrucomicrobia and Firmicutes decreased significantly in the 500 µg/L atrazine treatment group. At the genus level, Akkermansia and Lactococcus had significantly lower abundance in the 100 µg/L and 500 µg/L atrazine exposure group, while Cetobacterium was only detected in the 100 µg/L and 500 µg/L atrazine treated group. Also, function prediction of intestinal microbiota showed that atrazine treatment significantly changed the metabolism pathways of P. nigromaculatus tadpoles. In addition, 500 µg/L atrazine exposure changed the content of short chain fatty acids by significantly increasing the content of total SFCAs, butyric acid, and valeric acid, and decreasing the content of isovaleric acid in the intestine. Taken together, atrazine exposure could affect the intestinal histology and induce intestinal microbiota imbalance and metabolic disorder in amphibian tadpoles.

Individual and mixture toxicity of chromium and copper in development, oxidative stress, lipid metabolism and apoptosis of Bufo gargarizans embryos.

In natural ecosystems, living organisms are always subjected to a mixture of multiple heavy metals exposure, yet it is more common to study the effect of individual, rather than combined exposure. This study assessed the impacts of single or combined exposure to Cr and Cu on embryonic development, oxidative stress, lipid metabolism and apoptosis in the early development of Bufo gargarizans embryos. The total length, development stage and malformations of embryos were measured, and the mRNA expression of genes related to oxidative stress, lipid metabolism and apoptosis at Gs 18 and Gs 22 were determined by RT-qPCR. The results showed that all treatments significantly reduced the total length of embryos, delayed the stage of embryonic development and increased the proportion of malformed embryos. The Cr-Cu mixture treatment showed the greatest suppression of embryonic development and induced the highest rate of embryo malformation, compared to individual Cr and Cu treatments. In addition, the expression levels of oxidative stress genes (HSP90, SOD and GPx) and fatty acid ß-oxidation-related genes (ACOXL, ECHS1 and SCP) showed an up-regulated trend in treatments compared to control groups. Conversely, the lipid synthesis-related mRNA gene expressions (KAR, TECR, ACSL3 and ACSL4) were down-regulated. Among them, the Cr-Cu mixture had the greatest impact on lipid metabolism gene expression. The treatments showed significant effects on the expression of apoptosis genes (Bcl-1 and Bax), with Bcl-1 mRNA expression increasing and Bax mRNA expression decreasing. These results indicated that exposure to individual Cr, Cu and a Cr-Cu mixture can lead to oxidative stress, disrupt lipid metabolism and promote apoptosis, and the Cr-Cu mixture could cause more serious negative effects on B. gargarizans embryos than Cr or Cu individually.

The influence of long-term cadmium exposure on the male advertisement call of Xenopus laevis.

Cadmium (Cd) is a non-essential environmental endocrine-disrupting compound found in water and a potential threat to aquatic habitats. Cd has been shown to have various short-term effects on aquatic animals; however, evidence for long-term effects of Cd on vocal communications in amphibians is lacking. To better understand the long-term effects of low-dose Cd on acoustic communication in amphibians, male Xenopus laevis individuals were treated with low Cd concentrations (0.1, 1, and 10 µg/L) via aqueous exposure for 24 months. At the end of the exposure, the acoustic spectrum characteristics of male advertisement calls and male movement behaviors in response to female calls were recorded. The gene and protein expressions of the androgen receptor (AR) were determined using Western blot and RT-PCR. The results showed that long-term Cd treatment affected the spectrogram and formant of the advertisement call. Compared with the control group, 10 µg/L Cd significantly decreased the first and second formant frequency, and the fundamental and main frequency, and increased the third formant frequency. One and 10-µg/L Cd treatments significantly reduced the proportion of individuals responding to female calls and prolonged the time of first movement of the male. Long-term Cd treatment induced a downregulation in the AR protein. Treatments of 0.1, 1, and 10 µg/L Cd significantly decreased the expression of AR mRNA in the brain. These findings indicate that long-term exposure of Cd has negative effects on advertisement calls in male X. laevis.

Characterization of the complete mitochondrial genome of Zhenhai brown frog Rana zhenhaiensis (Anura: Ranidae).

Rana zhenhaiensis is a species of frog within the family Ranidae. In this study, its complete mitochondrial genome was characterized by high-throughput sequencing technology. It is 19,205 bp long with an overall AT content of 55.2% and includes 13 protein-coding genes (PCGs), 22 tRNA genes, 2 rRNA genes, and 1 control region. Besides, a 26-bp-long origin of L-strand replication (OL) is present between tRNA-Asn and tRNA-Cys. Our findings will be useful for the detailed study of mitogenome evolution and the phylogenetic relationships of the genus Rana and related taxa.

Intervention Effects of Atorvastatin Combined with Panax notoginseng Saponins on Rats with Atherosclerosis Complicated with Hepatic Injury.

BACKGROUND: Statins cannot be used for some active liver diseases, which limits its application to some extent. The combined use of statins with other drugs may be one of the ways to solve this dilemma. OBJECTIVE: This research aims to evaluate the effects of atorvastatin combined with Panax notoginseng saponins (PNS) on rats with atherosclerosis (AS) complicated with hepatic injury. MATERIALS AND METHODS: Seventy-two male Wistar rats were randomly categorized into control group (without any intervention, Group A) and AS model groups, which were divided into hepatic injury (Groups B-E) and nonhepatic injury (Groups F-I) groups. Hepatic and nonhepatic injury groups were intragastrically treated with 5.5 mg/kg·d atorvastatin (Group B, F), 200 mg/kg·d PNS (Group C, G), 5.5 mg/kg·d atorvastatin + 200 mg/kg·d PNS (Group D, H), and normal saline (Group E, I). After 8 weeks, total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol, low density lipoprotein-cholesterol (LDL-C), and serum calcium were analyzed to evaluate the hypolipidemic effect. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase, total bilirubin, and r-glutamyltransferase levels were measured to assess liver function. The thoracic aortas were used for hematoxylin-eosin staining. RESULTS: In both hepatic injury and nonhepatic injury groups, TC, TG and LDL-C levels significantly decreased in Groups B, D, F, and H. ALT and AST levels significantly increased in Group B, but significantly decreased in Groups C and D. The aortic intima thickness was significantly lower in Groups B, D, F, and H than that in the normal saline group. CONCLUSION: The combination of atorvastatin and PNS treatment showed a significant hypolipidemic effect and hepatic enzyme stability function. SUMMARY: The single use of Panax notoginseng saponins (PNS) in the rat model for atherosclerosis significantly reduced Ca2+ content in serum, whereas the effect of lowing total cholesterol (TC), triglyceride (TG), and low density lipoprotein-cholesterol (LDL-C) is not apparent, especially as compared with atorvastatin treatmentPNS combined with atorvastatin treatment of the rat model for atherosclerosis displayed a noticeable, synergistic effect that allowed for better reduction of TC, TG, LDL-C and Ca2+ in the serum than that with the single use of PNS or atorvastatinIn the rat liver injury combined with atherosclerosis model, the single use of PNS significantly improved liver function, whereas atorvastatin alone only aggravated liver injury in the rat model. The effect of PNS combined with atorvastatin on liver function was significantly better than that of atorvastatin aloneThe combined use of PNS and atorvastatin showed good stability of liver function on the liver injury combined with atherosclerosis model. Abbreviations used: PNS: Panax notoginseng saponins; AS: Atherosclerosis; TC: Total cholesterol; TG: Triglyceride; HDL-C: High density lipoprotein-cholesterol; LDL-C: Low density lipoprotein-cholesterol; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; ALP: Alkaline phosphatase; T-BIL: Total bilirubin; r-GT: R-glutamyltransferase; HE: Hematoxylin-eosin.

Pharmaceutical evaluation of naftopidil enantiomers: Rat functional assays in vitro and estrogen/androgen induced rat benign prostatic hyperplasia model in vivo.

Naftopidil (NAF) is a α1D/1A adrenoceptor selective drug used for the treatment of both benign prostatic hyperplasia and lower urinary tract symptoms (BPH/LUTS). However, NAF is used as a racemate in clinic. To compare the differences and similarities among two enantiomers and racemate, pharmacological activities were evaluated through rat functional assays in vitro and estrogen/androgen (E/T) induced rat BPH model in vivo. NAF and the two enantiomers showed similar blocking activity on α1 receptor. S-NAF exhibited more α1D/1A adrenoceptor subtype selectivity than R-NAF and the racemate. The selectivity ratios pA2 (α1D)/pA2 (α1B) and pA2 (α1A)/pA2 (α1B) were 40.7- and 16.2-fold, respectively. NAF and its enantiomers effectively prevented the development of rat prostatic hyperplasia via suppressing the increase of the prostatic wet weight, visually. The quantitative analysis of the relative acinus volume, relative stroma volume, relative epithelial volume, epithelial height and expression of proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA) were carried out. S-NAF showed an advantage on the effect of inhibiting prostate wet weight and stroma volume over R-NAF and racemate NAF (P<0.05). Nevertheless, no other significant difference was observed between these two enantiomers. In conclusion, both R-NAF and S-NAF not only relax prostate muscle but also inhibit the prostate growth, thus relieve BPH.

The potential effects of climate change on amphibian distribution, range fragmentation and turnover in China.

Many studies predict that climate change will cause species movement and turnover, but few have considered the effect of climate change on range fragmentation for current species and/or populations. We used MaxEnt to predict suitable habitat, fragmentation and turnover for 134 amphibian species in China under 40 future climate change scenarios spanning four pathways (RCP2.6, RCP4.5, RCP6 and RCP8.5) and two time periods (the 2050s and 2070s). Our results show that climate change may cause a major shift in spatial patterns of amphibian diversity. Amphibians in China would lose 20% of their original ranges on average; the distribution outside current ranges would increase by 15%. Suitable habitats for over 90% of species will be located in the north of their current range, for over 95% of species in higher altitudes (from currently 137-4,124 m to 286-4,396 m in the 2050s or 314-4,448 m in the 2070s), and for over 75% of species in the west of their current range. Also, our results predict two different general responses to the climate change: some species contract their ranges while moving westwards, southwards and to higher altitudes, while others expand their ranges. Finally, our analyses indicate that range dynamics and fragmentation are related, which means that the effects of climate change on Chinese amphibians might be two-folded.

The influence of low-dose cadmium on the laryngeal microstructure and ultrastructure of Pelophylax nigromaculata.

Cadmium (Cd) is one of the common heavy metals dispersed throughout the modern environment that disrupts the development of aquatic organisms. Amphibians appear to be particularly vulnerable to this heavy metal contaminant since their eggs and aquatic larvae live in aquatic habitats and have permeable skin. During this critical sensitive period, amphibians live primarily in the water and are thus susceptible to the effects of environmental pollutants in water. Pelophylax nigromaculata completes their laryngeal development from Gosner stage 19 (GS 19, embryonic stage) to GS 46 (metamorphosis stage). To study whether long-term (from GS 19 to GS 46) exposure to low Cd concentrations (0, 4, 8, 16, 32, 64, and 128 µg/L) affects the development of the larynx in P. nigromaculata, a comparative study of microstructures and ultrastructures of female and male P. nigromaculata larynges was carried out. In histological structure, the larynx was composed of epithelial tissue, skeletal muscle, and hyaline cartilage, and there were no differences between male and female frogs except that males had a larger cross-section area than females. In contrast to controls, 64 and 128 µg/L Cd treatments caused a significant decrease in cross-sectional area, while other treatments showed no significant differences. Under the scanning electron microscope, the surface of the larynx could be seen to be attached to developed and evenly distributed cilia, with no significant differences between young male and female frogs, even among the Cd treatments. Under transmission electron microscopy, developed laryngeal epithelial cells could be observed, with obvious cross striations of skeletal muscle cells and mitochondria distributed around the muscle and skeletal muscle satellite cells. Compared with the control group, mitochondria around the skeletal muscle grew in size and decreased in number in the high-concentration treatment.

Serum Procalcitonin and Procalcitonin Clearance as a Prognostic Biomarker in Patients with Severe Sepsis and Septic Shock.

We evaluated the tendency of the plasma concentration and procalcitonin (PCT) clearance (PCTc) to act as biomarkers of prognosis in patients with severe sepsis and septic shock. From 2011 to 2013, we prospectively analyzed patients with sepsis admitted to the intensive care unit (ICU). The serum PCT was evaluated at the time of sepsis diagnosis and again after 48 h (day 3) and 96 h (day 5). PCTc after 48 h (PCTc-day 3) and 96 h (PCTc-day 5) was also calculated to evaluate the prognostic value for survival in patients with sepsis. A total of 48 patients were included. Overall mortality was 16.7% (8 patients). PCTc was higher in survivors than in nonsurvivors, with significant differences on day 3 and day 5 (p = 0.033; p = 0.002, resp.); however, serum PCT levels on day 1, day 3, and day 5 were not significant prognostic factors for survival. The prognosis of patients with severe sepsis and septic shock may be associated with PCTc. Dynamic changes of PCT reflected as PCTc at 48 h (day 3) and 96 h (day 5) after admission to the ICU may serve as a predictor of survival in critically ill patients with severe sepsis.

The influence of long-term cadmium exposure on phonotaxis in male Pelophylax nigromaculata.

Cadmium (Cd) is a common industrial and agricultural heavy metal found in the natural environment that disrupts the endocrine systems of vertebrates. Amphibians are particularly vulnerable to endocrine disruptors because of their aquatic habitats and permeable skin. Endocrine disruptors are known to negatively affect amphibian acoustic behavior, but whether and how the ubiquitous pollutant Cd impacts this crucial amphibian signaling system remains unknown. Male black-spotted frogs (Pelophylax nigromaculata) show phonotactic responses to female receptive calls by emitting advertisement calls and moving towards females during the mating season, essential for reproductive success. To study whether long-term (60 d) exposure to low Cd concentrations (10(-8), 10(-7) and 10(-6) M) affects male phonotaxis, we recorded male responses to female calls following Cd exposure during the breeding season. We found that Cd adversely affected advertisement call characteristics (call latency, call duration and call rate), the proportion of individuals that responded and the time to first movement of the male. These results indicate that long-term exposure to Cd at environmentally relevant concentrations alters phonotaxis in male P. nigromaculata.

The predictive performance and stability of six species distribution models.

BACKGROUND: Predicting species' potential geographical range by species distribution models (SDMs) is central to understand their ecological requirements. However, the effects of using different modeling techniques need further investigation. In order to improve the prediction effect, we need to assess the predictive performance and stability of different SDMs. METHODOLOGY: We collected the distribution data of five common tree species (Pinus massoniana, Betula platyphylla, Quercus wutaishanica, Quercus mongolica and Quercus variabilis) and simulated their potential distribution area using 13 environmental variables and six widely used SDMs: BIOCLIM, DOMAIN, MAHAL, RF, MAXENT, and SVM. Each model run was repeated 100 times (trials). We compared the predictive performance by testing the consistency between observations and simulated distributions and assessed the stability by the standard deviation, coefficient of variation, and the 99% confidence interval of Kappa and AUC values. RESULTS: The mean values of AUC and Kappa from MAHAL, RF, MAXENT, and SVM trials were similar and significantly higher than those from BIOCLIM and DOMAIN trials (p<0.05), while the associated standard deviations and coefficients of variation were larger for BIOCLIM and DOMAIN trials (p<0.05), and the 99% confidence intervals for AUC and Kappa values were narrower for MAHAL, RF, MAXENT, and SVM. Compared to BIOCLIM and DOMAIN, other SDMs (MAHAL, RF, MAXENT, and SVM) had higher prediction accuracy, smaller confidence intervals, and were more stable and less affected by the random variable (randomly selected pseudo-absence points). CONCLUSIONS: According to the prediction performance and stability of SDMs, we can divide these six SDMs into two categories: a high performance and stability group including MAHAL, RF, MAXENT, and SVM, and a low performance and stability group consisting of BIOCLIM, and DOMAIN. We highlight that choosing appropriate SDMs to address a specific problem is an important part of the modeling process.

A new species of the genus Ctenophora Meigen (Diptera: Tipuloidea: Tipulidae) from China, with a key to the world species.

One new species of the genus Ctenophora Meigen, 1803, C. fumosa Men, sp. nov. (southern China: Anhui) is described and illustrated. A key to known species along with a checklist of known species of the genus Ctenophora are provided. The type specimens of the new species are deposited in the animal specimen room, School of Life Science, Anqing Normal University, Anqing, Anhui Province, China. 

Chronic effects of environmentally-relevant concentrations of lead in Pelophylax nigromaculata tadpoles: Threshold dose and adverse effects.

Lead (Pb) is a common heavy metal in the natural environment, but its concentration has been increasing alongside widespread industrial and agricultural development in China. The dark-spotted frog Pelophylax (formerly Rana) nigromaculata (Anura: Ranidae) is distributed across East Asia and inhabits anthropogenic habitats such as farmland. Here, P. nigromaculata tadpoles (Gosner stage 19-46) were exposed to Pb at different concentrations (0, 40, 80, 160, 320, 640 and 1280µg/L) and Pb-induced survival, metamorphosis time, development, malformations, mobility and gonad structure were monitored. The results showed that above the threshold concentration of Pb, adverse effects were obvious. As the concentration of Pb increased, the adverse effects on different traits followed different patterns: the effects on hindlimb length, survival rate, metamorphosis rate, total malformation rate, swimming speed and jumping speed largely exhibited a linear pattern; the effects on snout-vent length, body mass and forelimb length largely exhibited a bimodal pattern. Sex ratio and gonadal histology were not affected by Pb, suggesting that Pb is not strongly estrogenic in P. nigromaculata.

Significant change between primary and repeated serum laboratory tests at different time points in pediatric appendicitis.

OBJECTIVE: The aim of the work is to determine whether the change between primary and repeated serum inflammatory markers measured 8-12 h later may improve diagnostic accuracy in pediatric appendicitis. METHODS: The study group comprised 258 pediatric patients with clinically suspected appendicitis admitted to the pediatric emergency department from 2005 to 2007. The significant changes in serum parameters between primary and repeated examinations were selected as the discriminating variables. The receiver operating characteristic curves were used to determine the cut-off values of the changes between two examinations in predicting appendicitis. RESULTS: Receiver operating characteristic analysis showed that the cut-off values for the change in total neutrophils (3.0%) on the first day after the onset of symptoms (day 1), the changes in C-reactive protein concentration (4.5 mg/l) and in bands (1%) on day 2, and the change in C-reactive protein concentration (15.0 mg/l) on day 3 were significant serum parameters in predicting pediatric appendicitis. CONCLUSION: Repeated serum laboratory tests at different time points during the progression of acute appendicitis may be helpful in predicting pediatric appendicitis in the pediatric emergency department.

[Effects of cadmium on metamorphism and gonad differentiation in Rana chensinensis].

200 tadpoles of Rana chensinensis at stage 26 - 27 were exposed to 0.05, 0.1, 0.2 or 0.4 mg/L Cd2+ in tap water respectively until they're fully metamorphic after which the heteromorphic young frogs in different treatments were anatomized, females and males were identified through gonad observation, and the female ratio was calculated. Localization of estrogen receptors (ER) in liver cells was investigated in different treatments using immunocytochemistry. The results showed that Cd2+ might induce limb abnormality, however, there was little correlation between abnormality rate and cadmium concentration in lower Cd2+ levels except for a higher limb abnormality ratio in the 0.4 mg/L group. On the other hand, Cd2+ could affect gonad differentiation. Compared to the control group, the proportion of female population increased in the 0.05 mg/L group and decreased in the 0.1, 0.2 and 0.4 mg/L ones. The sex rate in the 0.2 mg/L group is significantly different from that in the control group. Hermaphrodite gonads appeared in the two treatments with 0.2 mg/L and 0.4 mg/L of Cd2+. Additionally, ER expression was positive in both cytoplasm and nucleolus of liver cells in Cd2+ treated groups. But, there was no linear relationship between ER expressions levels and the concentration of Cd2+. These results suggested that cadmium can influence tadpole metamorphosis and gonad development by affecting the secretion of sex hormone.

Impact of antibiotic use on fungus colonization in patients hospitalized due to fever.

Invasive candidiasis is an opportunistic infection that occurs in medical and surgical patients and carries a high mortality rate. Because its process always follows from colonization to amplification and to invasion, understand the colonization status is important to understand the likelihood of invasion. Screening for fungal colonization was performed with specimens from urine, throat, anus, and skin in 314 patients admitted for fever both before and after antibiotics treatment was administered. Throat (45%) and anus (43%) had the highest colonization rates. Only 7% of patient had fungal colonization on the skin. Candida albicans was the most frequently colonized species in throat (79%) and anus (70%). Colonized fungal species on skin were diverse, including Candida parapsilosis (33%), C. albicans (29%), and molds (24%). Sex (M:F ratio, 49.7:50.3) was not a factor in fungal colonization, but aging was associated with increased colonization rate. Forty-five patients received antibiotics treatment for more than 2 weeks and second surveillance cultures were taken at the end of the second week. Antibiotics treatment was associated with increased frequency of colonization (p=0.02), but the fungal species distribution pattern remained unchanged after antibiotic treatment.