A new species of Nanorana Günther, 1896 (Anura, Dicroglossidae) from Yunnan, China.

A new species of Nanorana Günther, 1896 is described from Yunnan Province, China, based on morphological and molecular evidence. Morphologically, Nanorana xuelinensis sp. nov. is distinguished from its congeners by a combination of the following diagnostic characters: body size large; adult males with keratinized spines on chest, belly, lateral body, posterior dorsum, buttocks, outer side of the fore limbs, the inner metacarpal tubercle, fingers I and II, and upper eyelids; no spines on the inner side of the lower and upper arm; forelimbs strongly hypertrophied in adult males; anterior dorsum skin smooth; dorsolateral folds absent; finger I longer than finger II; webbing deeply incurved between tips of toes; present outer metacarpal tubercle and absent outer metatarsal tubercle. The new species is separated from all other congeners by uncorrected genetic distances ranging from 5.2% to 7.3% based on mitochondrial 16S rRNA gene and ranging from 3.9% to 7.6% based on mitochondrial 12S rRNA gene.

Downregulating lncRNA PVT1 Relieves Astrocyte Overactivation Induced Neuropathic Pain Through Targeting miR-186-5p/CXCL13/CXCR5 Axis.

Spinal cord injury (SCI) is one of the main causes leading to neuropathic pain. Here, we aim to explore the molecular mechanism and function of lncRNA PVT1 in neuropathic pain induced by SCI. The expression of lncRNA PVT1, microRNA (miR) - 186-5p was measured via quantitative reverse transcription PCR (qRT-PCR), and the activation of astrocytes (labeled by GFAP) was detected by immunohistochemistry. Western blot was conducted to detect the expression of chemokine ligand 13 (CXCL13), chemokine receptor 5 (CXCR5), cyclooxygenase-2 (COX2), inducible nitric oxide synthase (iNOS) and glial fibrillary acidic protein (GFAP) in spinal cord injury lesions. The levels of inflammatory cytokines (including IL-1ß and IL-6) and MDA in tissues were examined via Enzyme-linked immunosorbent assay (ELISA). In vitro experiments were also conducted in primary cultured astrocyte to explore the response of astrocyte to lipopolysaccharide (LPS). What's more, the PVT1-miR-186-5p interaction was verified via the dual luciferase activity assay and RNA immunoprecipitation (RIP) assay. The results demonstrated that the levels of PVT1, CXCL13 and CXCR5 were upregulated, while miR-186-5p were decreased in SCI rats' spinal cord and LPS-mediated astrocytes. In the SCI model, PVT1 depletion significantly alleviated neuropathic pain, astrocytic activation and reduced the expression of neuroinflammatory factors and proteins. The relevant mechanism studies confirmed that PVT1 is a competitive endogenous RNA (ceRNA) of miR-186-5p, targets and inhibits its expression and promotes the expression of CXCL13/CXCR5, while miR-186-5p targets CXCL13. In conclusion, inhibition of lncRNA PVT1 alleviates neuropathic pain in SCI rats by upregulating miR-186-5p and down-regulating CXCL13/CXCR5. The PVT1/miR-186-5p/CXCL13/CXCR5 axis can be used as a new therapeutic target for neuropathic pain.

Response of bacterial communities in rubber plantations to different fertilizer treatments.

In the present study, the effects of chemical fertilizer (CF) and organic fertilizer plus chemical fertilizer application (OF-CF) on natural rubber yield, soil properties, and soil bacterial community were systematically investigated in rubber plantations. The rubber dry yield was 26.3% more in the OF treatment group than in the CF treatment group. The contents of total nitrogen (TN), available nitrogen (AN), available phosphorus (AP), and available potassium (AK) as well as soil organic matter (SOM) and pH value were higher following OF-CF treatment. Using Illumina sequencing, a total of 927 operational taxonomic units (OTUs) were obtained following CF treatment, while 955 OTUs were obtained after OF-CF treatment. Relative abundance analysis showed the relative abundances of four phyla (Acidobacteria, Proteobacteria, Actinobacteria, Gemmatimonadetes) were different between the two treatment groups. Correlation analysis revealed Acidobacteria, Bacteroidetes, Thaumarchaeota, Elusimicrobia, Verrucomicrobia were the key taxa that determined the soil properties. Additionally, five OTUs (OTU_506, OTU_391, OTU_189, OTU_278, OTU_1057) were thought to be related to the biodegradation of natural rubber. Taken together, these results improve our understanding of the OF-mediated improvement in soil fertility and contribute to the identification of rubber-degrading bacteria in rubber plantations.

Randomized trial of umbilical incisional hernia in high-risk patients: extraction of gallbladder through subxiphoid port vs. umbilical port after laparoscopic cholecystectomy.

INTRODUCTION: Trocar site incisional hernia (TSIH) is one of the most common complications of laparoscopic surgery. Using the umbilical port as a common hole for removing the gallbladder in laparoscopic cholecystectomy is more likely to lead to TSIH than other ports. Thus, extracting the gallbladder through other ports may reduce the incidence of TSIH. AIM: To ascertain whether extraction of the gallbladder through the subxiphoid port is more beneficial for reducing umbilical incisional hernia than the umbilical port. MATERIAL AND METHODS: From April 2014 to March 2017, a randomized clinical trial was conducted among patients with high risk of incisional hernia and accepted for three-port laparoscopic cholecystectomy (TLC) in our department. 182 patients with indications of cholecystectomy were allocated randomly to group A (subxiphoid port) and group B (umbilical port). Data collection was carried out on operative time, postoperative pain, hospital stay, wound infection and TSIH in the early postoperative course, and at 1, 10, and 24 months after surgery. RESULTS: The incidence of TSIH in group A was lower than that in group B (4.9% vs. 14.6%; odds ratio = 8.02; 95% CI: 2.15-47.6; p < 0.001). The mean operative time of group A was significantly shorter than that of group B (35 ±15.16 min vs. 42 ±14.58 min, p < 0.01). There was no significant difference in wound infection rate between group A and group B (p = 0.068). The data of hospital stay (p = 0.428) and postoperative pain (p = 0.349) of all analyzed patients were similar in the two groups. CONCLUSIONS: Extraction of the gallbladder through the subxiphoid port can reduce umbilical incisional hernia in high-risk patients effectively.

FOXK1 promotes cell growth through activating wnt/ß-catenin pathway and emerges as a novel target of miR-137 in glioma.

Glioma is the most common primary malignant brain tumor in adults. Forkhead box k1 (FOXK1) was reported to be dysregulated and play important roles in multiple human cancers. However, the expression pattern and roles of FOXK1 in glioma has never been investigated. In this study, we firstly observed that the expression of FOXK1 was significantly increased in glioma tissue samples and cell lines. Functional assays demonstrated that overexpression of FOXK1 promoted proliferation, cell cycle transition and inhibited apoptosis in glioma cell lines. On the contrary, knockdown of FOXK1 exhibited an opposite effect on glioma cells proliferation, cell cycle and apoptosis. Data of western blot indicated that FOXK1 overexpression increased while FOXK1 knockdown decreased the levels of ß-catenin, c-myc and cyclinD1 in glioma cells. Moreover, we demonstrated that FOXK1 was a novel target of miR-137 in glioma and FOXK1 restoration abolished the tumor suppressive effect of miR-137 in glioma cells. Statistical analysis showed that the mRNA level of FOXK1 was inversely correlated with miR-137 expression in glioma tissues. In conclusion, the present study demonstrated that FOXK1 promoted cell growth through activating wnt/ß-catenin pathway and is negatively regulated by miR-137 in glioma.

Heart rate changes in partial seizures: analysis of influencing factors among refractory patients.

BACKGROUND: We analyzed the frequency of heart rate (HR) changes related to seizures, and we sought to identify the influencing factors of these changes during partial seizures, to summarize the regularity of the HR changes and gain some insight into the mechanisms involved in the neuronal regulation of cardiovascular function. To date, detailed information on influencing factors of HR changes related to seizures by multiple linear regression analysis remains scarce. METHODS: Using video-electroencephalograph (EEG)-electrocardiograph (ECG) recordings, we retrospectively assessed the changes in the HR of 81 patients during a total of 181 seizures, including 27 simple partial seizures (SPS), 110 complex partial seizures (CPS) and 44 complex partial seizures secondarily generalized (CPS-G). The epileptogenic focus and the seizure type, age, gender, and sleep/wakefulness state of each patient were evaluated during and after the seizure onset. The HR changes were evaluated in the stage of epilepsy as time varies. RESULTS: Of the 181 seizures from 81 patients with ictal ECGs, 152 seizures (83.98%) from 74 patients were accompanied by ictal tachycardia (IT). And only 1 patient was accompanied by ictal bradycardia (IB). A patient has both IT and IB. We observed that HR difference was independently correlated with side, type and sleep/wakefulness state. In this analysis, the HR changes were related to the side, gender, seizure type, and sleep/wakefulness state. Right focus, male, sleep, and CPS-G showed more significant increases than that were observed in left, female, wakefulness, SPS and CPS. HR increases rapidly within 10 seconds before seizure onset and ictus, and typically slows to normal with seizure offset. CONCLUSION: CPS-G, sleep and right focus led to higher ictal HR. The HR in the stage of epilepsy has regularly been observed to change to become time-varying. The risk factors of ictal HR need to be controlled along with sleep, CPS-G and right focus. Our study first explains that the HR in seizures has a regular evolution varying with time. Our study might help to further clarify the basic mechanisms of interactions between heart and brain, making seizure detection and closed-loop systems a possible therapeutic alternative in refractory patients.

Improvements of immune status, intestinal integrity and gain performance in the early-weaned calves parenterally supplemented with L-alanyl-L-glutamine dipeptide.

Glutamine (Gln) is an important substrate for the innate immune cells including lymphocytes and macrophages. In this report, the effects of alanyl-glutamine dipeptide (Ala-Gln) on the naïve immune system, intestinal integrity and gain performance of early-weaned calves were investigated. Early-weaned Chinese Holstein calves were intravenously administered different dosages of Ala-Gln. The effects of Ala-Gln administration were ascertained by evaluating the blood for naïve T lymphocyte subpopulations, the concentrations of serum IgG, serum IgA and intestinal mucosal secretory IgA (s-IgA), the intestinal integrity, as well as the gain performance. Results demonstrated that intravenous administration of Ala-Gln dipeptide (1.01 g/kg×d(-1)) for 7 days had a positive effect on gain performance, intestinal integrity and the immune system. Calves administered doses of Ala-Gln displayed an improvement in gain performance and health status concurrent with increases in blood CD2(+) and CD4(+) lymphocytes, the ratio of CD4(+)/CD8(+), serum IgA and IgG, intestinal mucosal s-IgA while decreasing the occurrence of diarrhea. Moreover, we found that animals given the effective dose (1.01 g/kg×d(-1)) of Ala-Gln resulted in improved immune status and intestinal integrity relative to those given a lower (0.49 g/kg×d(-1)) or higher dose (1.99 g/kg×d(-1)) of Ala-Gln. These findings suggest that maintaining a certain concentration of plasma and/or tissue glutamine in the early stages of weaning is an effective alternative approach for improvement of growth performance in early-wean calves.

[Expression of neuropeptides ghrelin and Nesfatin-1 in kainic acid kindling rats].

OBJECTIVE: To observed the post-seizure expression level of neuropeptides ghrelin and Nesfatin-1 between the untreated and treated groups in kainic acid-kindling rats and understand the significance of their basic expressions. METHODS: The male S-D rats were divided randomly into NS and VPA groups after seizure. Each group was divided into 7 subgroups of 3 h, 6 h, 12 h, 24 h, 3 d, 7 d and 14 d. And the blank group (without NS or VPA) and sham group (normal saline instead of kainic acid) were established. RIA (radioimmunoassay), ELISA (enzyme-linked immunosorbent assay) and immunohistochemistry were used to detect the expression levels of ghrelin and Nesfatin-1 in the serum and hypothalamus of kainic acid-kindling rats. RESULTS: Compared with the blank and sham groups, the seizure rats had a decreased expression of ghrelin while there was an elevated expression of Nesfatin-1 in hypothalamus. This trend was consistent with the serum expressions of deacylated ghrelin and Nesfatin-1. The expression of ghrelin became elevated at all stages in the VPA group. Especially the subgroups of 24 h (0.320 ± 0.007) and 7 d (0.284 ± 0.004) had a significant increase (P < 0.05) while their expression levels remained lowed than that of the blank group (0.342 ± 0.039). The expressions of Nesfatin-1 had a significant decrease (P < 0.05) in all VPA subgroups while they were higher than those in the blank group (0.124 ± 0.007). CONCLUSION: An accurate detection of serum expression level of deacylated ghrelin and Nesfatin-1 may reflect the fluctuation trend of neuropeptides in hypothalamus. It may offer a simple, sensitive and noninvasive method of diagnosing and treating epilepsy.

[mRNA expression of activity-regulated cytoskeletal-associated protein in hippocampus induced by insular-kindled rats].

OBJECTIVE: To investigate the expression of Arc mRNA in insular electrical kindled and single electrical stimulated rats and its significance. METHODS: Male SD rats were divided randomly into kindled group, single electrical stimulated group, sham-operated group and control group. Each group was divided into 2 sub-groups at different time points. Kindled group: establishing chronic insular electrical kindled model, decapitation to perform RT-PCR of Arc mRNA on hippocampus and to applying in situ hybridization of Arc mRNA on dentate gyrus; Single electrical stimulated group: using the same method as the kindled group with only electrical stimulation twice; Sham-operated group: using the same method to the kindled group without electrical stimulation; CONTROL GROUP: no surgery. RESULTS: Expression of Arc mRNA in the hippocampus of single electrical stimulated, sham-operated and control groups were 0.72 ± 0.14, 0.75 ± 0.16 and 0.71 ± 0.14 respectively. They were significantly less than the kindled group of 1.78 ± 0.43(P < 0.01) at 3 h. The expression of Arc mRNA had no significant difference (P > 0.05) among 4 groups at 6 h; in situ hybridization (cell count) of Arc mRNA showed the expression of Arc mRNA in the kindled group was 112.8 ± 6.0. And it was significantly higher than the other three groups of 46.25 ± 4.35, 45.25 ± 6.23, 44.75 ± 6.49 (P < 0.01) after 3 h, there were no significant difference among 4 groups at 6 h (P > 0.05). CONCLUSION: Insular epilepsy increase the expression of Arc mRNA in hippocampus. Arc may play an important role in the synaptic plasticity of insular epilepsy.

[Expressions of GAP43, P38 mRNA and protein in insular electrical kindled rats and its significance].

OBJECTIVE: To investigate the expression of neural growth-associated protein (GAP-43) and synaptophysin (P38) gene and protein in insular electrical kindled epileptic rats and its significance. METHODS: Seventy-two male adult rats were divided into kindled group (n = 36) and sham group (n = 36) randomly. Each group was further divided into 1, 3, 7, 15, 30 and 60 d sub-groups (n = 6) respectively. After kindling with immunohistochemical staining and hybridization in situ, the altered expressions of GAP-43 and P38 were detected in hippocampus of insular electrical kindled epilepsy rats. RESULTS: Epilepsy day 1: GAP-43 mRNA and protein expression at hippocampus dentate gyrus granular cell layer, hilus and CA3 pyramidal layer in the kindled group were higher than the sham group (P < 0.05). The expressions declined at Day 3 (P > 0.05). Expressions became elevated again at Day 7 and they were significantly higher than those in the sham group at Days 15 and 30 (P < 0.05). There was no statistical difference between two groups at Day 60 (P > 0.05). Expressions of P38 mRNA and protein: hybridization signal and immunostaining at hippocampus dentate gyrus granular cell layer, hilus and CA3 pyramidal layer began to rise at Day 7 and peaked at Day 15 (P < 0.01). They lasted 4 weeks and then began to decline. CONCLUSION: Insular epilepsy is closely related with temporal hippocampus. GAP-43 and P38 may be the pathological basis of insular epilepsy and the key molecular mechanism of synaptic plasticity.