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Cisplatin, also known as cis-diamine dichloroplatinum (CDDP), is a widely used chemotherapeutic drug. However, its application is limited by the occurrence of serious nephrotoxicity. Currently, no effective therapy is available for combating CDDP-induced acute kidney injury (AKI). In the present study, we investigated the efficacy of Jianpi Yishen Tang (JPYST), a traditional Chinese medicine (TCM) compound commonly used to treat chronic kidney disease, against CDDP-induced AKI. In the CDDP + JPYST group, male mice were pretreated with JPYST (18.35 g/kg/day) for 5 consecutive days before receiving a single dose of CDDP (20 mg/kg), all mice were sacrificed 72 h after the CDDP injection. Results showed that JPYST suppressed CDDP-induced kidney dysfunction and tubular damage scores in the mice. Mechanistically, JPYST treatment attenuated CDDP-induced renal tubular cell apoptosis in AKI mice, as manifested by a marked decreased in TUNEL-positive cell counts, downregulation of the pro-apoptotic proteins Bax, Bad and caspase 3, and upregulation of the antiapoptotic protein Bcl-2 in kidney tissues. Meanwhile, JPYST decreased the expression of inflammatory cytokines TNF-α, IL-1β, and IL-6 in the serum and renal tissues of mice following CDDP administration. These factors are involved in suppressing the activation of phospho-NF-κB p65 in tubular epithelial cells. Taken together, these findings demonstrated that JPYST exerts renoprotective effects against CDDP-induced AKI through antiapoptosis and anti-inflammation effects, and these are associated with downregulation of NF-κB activation. Therefore, JPYST has potential for development of treatment therapies against CDDP-induced AKI.

N-Methyl-d-aspartate receptors (NMDARs) are glutamate-gated Na+ and Ca2+-permeable ion channels involved in excitatory synaptic transmission and synaptic plasticity. NMDAR hypofunction has long been implicated in the pathophysiology including major depressive disorders (MDDs). Herein, we report a series of furan-2-carboxamide analogues as novel NMDAR-positive allosteric modulators (PAMs). Through structure-based virtual screen and electrophysiological tests, FS2921 was identified as a novel NMDAR PAM with potential antidepressant effects. Further structure-activity relationship studies led to the discovery of novel analogues with increased potentiation. Compound 32h caused a significant increase in NMDAR excitability in vitro and impressive activity in the forced swimming test. Moreover, compound 32h showed no significant inhibition of hERG or cell viability and possessed a favorable PK/PD profile. Our study presented a series of novel NMDAR PAMs and provided potential opportunities for discovering of new antidepressants.

Current pain management is largely limited to opioids and non-steroidal anti-inflammatory drugs. Developing new analgesic drugs remains important to address the unmet medical needs of chronic pain patients. Calcium-activated chloride channel anoctamin-1 (ANO1) is a potential analgesic target. ANO1 is activated by noxious stimuli in peripheral sensory neurons and further induced neural depolarization. Downregulation of ANO1 reduced hyperalgesia and allodynia caused by inflammation and nerve injury. Here we developed a series of 4-arylthiophene-3-carboxylic acid derivatives for proof-of-concept studies of ANO1-targeted analgesia. These efforts led to the identification of the compound DFBTA, 4-(4-chlorophenyl)-2-(2,5-difluorobenzamido)thiophene-3-carboxylic acid, which displays dramatic ANO1 inhibition with IC

In this study, a high-pressure membrane reactor (MR) was employed to carry out the methanol synthesis (MeS) reaction. Syngas was fed into the MR shell side where a commercial MeS catalyst was used, while the tube side was swept with a high boiling point liquid with good solubility toward methanol. A mesoporous alumina ceramic membrane was utilized after its surface had been modified to be rendered more hydrophobic. The efficiency of the MR was investigated under a variety of experimental conditions (different pressures, temperatures, sweep liquid flow rates, and types of sweep liquids). The results reveal improved per single-pass carbon conversions when compared to the conventional packed-bed reactor. An ionic liquid (IL), 1-ethyl-3-methylimidazolium tetrafluoroborate, was utilized in the MR as the sweep liquid. The experimental results are compared to those previously reported by our group (Li, Z.; Tsotsis, T. T. J. Membrane Sci. 2019, 570, 103) while using a conventional petroleum-derived solvent as swe...

In order to improve the quality in chemical mechanical polishing (CMP) of stainless steel and the material removal rate (MRR), 6 kinds of abrasives were used to do the CMP test of 304 stainless steel. In this paper, the 6 kinds of abrasives have different hardness. The experimental results show that, in the CMP process, the MRR increases with the increase of the abrasive hardness, the surface roughness decreases firstly and then raises with the increase of the abrasive hardness. This surface roughness reduces the lowest value when the chromium oxide (CrO) abrasive is used. With low-hardness copper oxide abrasives and magnesium oxide abrasives, it can only remove partial scratches on the surface during the polishing process and obtained low MRR and poor surface quality. Silica abrasives were easy to condense in the polishing slurry and the Gelation hindered the cutting action of abrasive, So resulting in low MRR and poor surface quality. The chromium oxide and white corundum, their hardness are close to the stainless steel, can remove the surface oxide layer of the sample while not produce the damage of the workpiece surface, so the surface quality was very well. Considering that chromium oxide contains Cr2+, it will pollute the environment. Therefore, in comprehensive consideration of the surface quality and MRR, take the white corundum as an abrasive in the CMP of 304 stainless steel.

Background Hepatic ischemia-reperfusion (I/R) injury (IRI) represents a crucial challenge in liver transplantation. Fisetin has anti-inflammatory, anti-aging and anti-oxidative properties. This study aimed to examine whether fisetin mitigates hepatic IRI and examine its underlying mechanisms. Methods Sham or warm hepatic I/R operated mice were pretreated with fisetin (5, 10 or 20 mg/kg). Hepatic histological assessments, TUNEL assays and serum aminotransferase measurements were performed. An in vitro hypoxia/reoxygenation (H/R) model using RAW264.7 macrophages pretreated with fisetin (2.5, 5 or 10 µmol/L) was also used. Serum and cell supernatant concentrations of interleukin-1β (IL-1β), IL-18 and tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA). Protein levels of p-GSK3β, p-AMPK and NLR family pyrin domain-containing 3 (NLRP3)-associated proteins were detected by Western blotting. Results Compared with the I/R group, fisetin pretreatment reduced pathological liver damage, serum aminotransferase levels, serum concentrations of IL-1β, IL-18 and TNF-α in the murine IRI model. Fisetin also reduced the expression of NLRP3 inflammasome-associated proteins (NLRP3, cleaved caspase-1, IL-1β and IL-18) in I/R-operated liver. The experiments in vitro showed that fisetin decreased the release of IL-1β, IL-18 and TNF-α, and reduced the expression of NLRP3 inflammasome-associated proteins in H/R-treated RAW264.7 cells. Moreover, fisetin increased the expressions of p-GSK3β and p-AMPK in both models, indicating that its anti-inflammatory effects were dependent on GSK3β/AMPK signaling. The anti-inflammatory effects of fisetin were partially inhibited by the AMPK specific inhibitor compound C. Conclusions Fisetin showed protective effects against hepatic IRI, countering inflammatory responses through mediating the GSK3β/AMPK/NLRP3 inflammasome pathway.

Ataxia-telangiectasia mutated (ATM) kinase is a serine/threonine protein kinase and plays a key role in DNA double-strand breaks repair. Thus, ATM is considered a promising target for radiotherapy and chemotherapy sensitizing. Herein, we report the discovery of ATM agonist A22 and inhibitor A41 by computational methods and further biological evaluation. Among them, A22 exhibited low cytotoxicity in vitro and might serve as a useful tool for ATM research. Moreover, we firstly proved that ATM inhibitors could sensitize Irinotecan and Etoposide in a time-dependent manner on MCF-7 and SW480 cells, antagonism in a short period treatment while synergy at a long-term treatment and ATM agonist worked in an opposite way of ATM inhibitors. Further mechanism study demonstrated that the antagonism effect of ATM inhibitors with chemotherapeutic agents in a short period was resulting from inhibiting the p53/p21 axis to accelerate G1/S phase cell-cycle transition and promote cell survival. Additionally, A41 displayed antitumor effects combined with a chemotherapeutic drug in the SW480 xenograft model, indicating that A41 is a promising ATM inhibitor, which could increase the antitumor effect of chemotherapeutic drugs in vivo. All in all, these findings will guide the combination of ATM inhibitors with chemotherapeutic agents in further preclinical and clinical studies.

International audience; A series of organoselenium compounds based on the hybridization of nonsteroidal antiinflammatory drugs (NSAIDs) scaffolds and Se functionalities (-SeCN and -SeCF3) were synthesized and characterized, and evaluated against four types of cancer cell lines, SW480 (human colon adenocarcinoma cells), HeLa (human cervical cancer cells), A549 (human lung carcinoma cells), MCF-7 (human breast adenocarcinoma cells). Interestingly, most of the investigated compounds showed active in reducing the viability of different cancer cell lines. The most active compound 3h showed IC50 values lower than 20 μM against the four cancer cell lines, particularly to SW480 and MCF-7 with IC 50 values of 4.9 and 3.4 μM, respectively. Furthermore, NSAIDs-SeCN derivatives (2h and 2i) and NSAIDs-SeCF3 derivatives (3h and 3i) were selected to investigate their ability to induce apoptosis in MCF-7 cells via modulation the expression of anti-apoptotic Bcl-2 protein, pro-inflammatory cytokines (IL-2) and proapoptotic caspase-3 protein. Moreover, the redox properties of the synthesized organoselenium candidates were conducted by 2, 2-didiphenyl-1-picrylhydrazyl (DPPH), bleomycin dependent DNA damage and glutathione peroxidase (GPx)-like assays. Taken together, these NSAIDs-Se candidates could provide promising new lead derivatives for further potential anticancer drug development.

A series of organoselenium compounds based on the hybridization of nonsteroidal antiinflammatory drugs (NSAIDs) scaffolds and Se functionalities (-SeCN and -SeCF

Hepatitis B virus (HBV) causes serious health issues worldwide. Despite this, current treatment options for HBV have many drawbacks. Strategies to safely and specifically target HBV replication and survival at the transcriptional level within host cells are needed to combat current drawbacks in treatment. In this study, we designed a novel artificial transcription factor (ATF) with suppressive function to target and bind to the HBV core promoter, a component that plays a central role in the viral life cycle. ATF has attached specifically to the intended target site by using electrophoretic mobility shift assays (EMSA). We tested whether targeting this suppressive ATF had any effect on HBV gene expression by transfection factor, western blotting, and real-time PCR. In the presence of ATF, viral mRNA and DNA levels were significantly decreased within HepG2.2.15 cells compared to control cells. The HBV-derived protein expression of HBV-e antigen (HBeAg) and HBV-c antigen (HBcAg) was also significantly inhibited. These results show that ATF treatment targeting the HBV core protein promoter has an antiviral effect and inhibits HBV infection in host cells. These results further suggest that the design of new artificial transcription factors may be valuable antiviral therapies to treat HBV patients.

Dexmedetomidine (Dex) protects different cell types during hypoxia or ischemia-reperfusion injury by inhibiting cell apoptosis. However, the underlying mechanism and its impact on hepatic ischemia reperfusion injury are still not known. In this study, we established a model of oxygen-glucose deprivation/reperfusion (OGD/R) injury in hepatocyte HL7702 cells, and studied the impact of Dex on cell proliferation, apoptosis, and cell cycle during OGD/R. In addition, we explored the role of CCAT1 in this process. We found that Dex increased cell proliferation and inhibited cell apoptosis during OGD/R, in a concentration-dependent manner. Dex partially reversed the OGD-inhibited expression of lncRNA CCAT1. Knockdown of CCAT1 by siRNA inhibited Dex-mediated protection against OGD/R-induced injury and promoted cell apoptosis, caspase-3 expression and cell cycle arrest in the G0/G1 phase, and inhibited cell proliferation and cyclin D1 expression. In contrast, overexpression of CCAT1 by pcDNA3.0-CCAT1 enhanced Dex-mediated protection against OGD/R-induced cell injury. Thus, Dex protects hepatocytes against OGD/R injury by upregulating lncRNA CCAT1. This study suggests a novel role of CCAT1 in ischemia reperfusion injury, and lays the framework for future studies.

Background Recent studies have demonstrated that IL-34 is implicated in the regulation of macrophage functions. However, the effect of IL-34 on Kupffer cells (KCs) in acute rejection (AR) of liver transplantation remains unclear. Methods IL-34 expression was detected in graft and serum from allotransplantation and syngeneic transplantation groups. The adeno-associated virus-expressing IL-34 was used to assess the effect of IL-34 on AR of rat liver transplantation. The effect of IL-34 on KC polarization was evaluated by in vitro and in vivo assays. Kupffer cells in donors were depleted by clodronate treatment before transplantation, and the nontreated KCs or lipopolysaccharide-treated KCs were transferred into recipients during liver transplantation. Results IL-34 expression levels in grafts and serum were decreased in the allotransplantation group compared with the syngeneic transplantation group. Adeno-associated virus-expressing IL-34 treatment induced KC M2 polarization in vivo and inhibited the AR of rat liver transplantation. Moreover, we found that IL-34 switched the phenotype of KCs from M1 to M2 by activating the PI3K/Akt pathway in vitro. In addition, the results of KC deletion and adaptive transfer experiments showed that the AR inhibition induced by IL-34 was M2 KC-dependent. Conclusions IL-34 plays an important role in KC M2 polarization-dependent AR inhibition of rat liver transplantation.

Background Papillary renal cell carcinoma (PRCC) is the most common type of renal cell carcinoma after clear cell renal cell carcinoma (ccRCC). Its pathological classification is controversial, and its molecular mechanism is poorly understood. Therefore, the identification of key genes and their biological pathways is of great significance to elucidate the molecular mechanisms of PRCC occurrence and progression. Methods The PRCC-related datasets GSE7023, GSE48352 and GSE15641 were downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were identified, and gene ontology (GO) term enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed. Cytoscape and STRING were used to construct the protein-protein interaction network (PPI) and perform module analysis to identify hub genes and key pathways. A heatmap of hub genes was constructed using the UCSC cancer genomics browser. Overall survival and recurrence-free survival of patients stratified by the expression levels of hub genes were analysed using Kaplan-Meier Plotter. The online database UALCAN was applied to analyse gene expression based on tissue type, stage, subtype and race. Results A total of 214 DEGs, specifically, 205 downregulated genes and 9 upregulated genes, were identified. The DEGs were mainly enriched in angiogenesis, kidney development, oxidation-reduction process, metabolic pathways, etc. The 17 hub genes identified were mainly enriched in the biological processes of angiogenesis, cell adhesion, platelet degranulation, and leukocyte transendothelial migration. Survival analysis showed that EGF, KDR, CXCL12, REN, PECAM1, CDH5, THY1, WT1, PLAU and DCN might be related to the carcinogenesis, metastasis or recurrence of PRCC. UALCAN analysis showed that low expression of PECAM1 and PLAU in PRCC tissues was related to stage, subtype and race. Conclusions The DEGs and hub genes identified in the present study provide insight into the specific molecular mechanisms of PRCC occurrence and development and may be potential molecular markers and therapeutic targets for the accurate classification and efficient diagnosis and treatment of PRCC.

Aims Hepatic ischemia/reperfusion (I/R) injury is a critical factor affecting the prognosis of liver surgery. The aim of this study is to explore the effects of SET8 on hepatic I/R injury and the putative mechanisms. Main methods The expression of SET8 and MARK4 in I/R group and sham group were detected both in vivo and in vitro. In addition, mouse and RAW 264.7 cells were transfected with MARK4 siRNA and SET8 siRNA knockdown of MARK4 and SET8, respectively. The expression of SET8, MARK4 and NLRP3-associated proteins were detected after different treatments. The pathology of liver and the serologic detection were detected after different treatments. Key findings Our present study identified SET domain-containing protein 8 (SET8) as an efficient protein, which can negatively regulate hepatic I/R-mediated inflammatory response and ameliorate hepatic I/R injury by suppressing microtubule affinity-regulating kinase 4 (MARK4)/ NLR family pyrin domain containing 3 (NLRP3) inflammasome pathway. The data showed that MARK4 deficiency inhibited hypoxia/reoxygenation (H/R)-induced NLRP3 inflammasome activation, while SET8 deficiency showed the opposite effect. We further demonstrated that SET8 restrained NLRP3 inflammasome activation by inhibiting MARK4. Moreover, we verified SET8 made protective effect on hepatic I/R injury. Significance SET8 plays an essential role in hepatic ischemia/reperfusion injury in mice by suppressing MARK4/NLRP3 inflammasome pathway. Our results may offer a new strategy to mitigate hepatic I/R injury.

// Rui Liu 1 , Chengyong Tang 2 , Ai Shen 1, 3 , Huating Luo 4 , Xufu Wei 1 , Daofeng Zheng 1 , Chao Sun 1 , Zhongtang Li 1 , Di Zhu 1 , Tingting Li 1 , Zhongjun Wu 1 1 Department of Hepatobiliary Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China 2 Department of Clinical Pharmacology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China 3 Department of Hepatobiliary Surgery, Chongqing Cancer Institute, Chongqing 400030, China 4 Department of Oncology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China Correspondence to: Zhongjun Wu, email: wzjtcy@126.com Keywords: hepatocellular carcinoma, IL-37, TGF-β, JNK/pSmad3L/c-Myc, pSmad3C/p21 Received: April 01, 2016 Accepted: October 26, 2016 Published: November 08, 2016 ABSTRACT IL-37 has been characterized as a fundamental inhibitor of innate immunity and a tumor suppressor in several cancers. However, the molecular mechanism of IL-37 in hepatocellular carcinoma (HCC) is largely unclear. In this study we found IL-37 expression was down-regulated in human HCC tissues and cell lines, and was negatively correlated with tumor size, vascular invasion, as well as overall-survial and disease-free survival (OS and DFS) of HCC. Multivariate Cox analysis revealed that IL-37 was an independent prognostic indicator for OS and DFS in HCC. Functional studies further showed that IL-37 overexpression significantly suppressed tumor growth by confining HCC to G2/M cell cycle arrest in vitro and in vivo . Mechanistically, we determined that IL-37 promoted Smad3 phospho-isoform signaling conversion from JNK/pSmad3L/c-Myc oncogenic signaling to pSmad3C/p21 tumor-suppressive signaling. Consistently, we detected a significant negative correlation between IL-37 expression and pSmad3L levels in a cohort of HCC biopsies; and the expression of pSmad3L predicted poorer outcome. These data highlight the importance of IL-37 in the cell proliferation and progression of HCC, and suggests that IL-37 may be a valuable biomarker for HCC prognosis.

Summary: cADPR is a well-recognized signaling molecule by modulating the RyRs, but considerable debate exists regarding whether cADPR can bind to and gate the TRPM2 channel, which mediates oxidative stress signaling in diverse physiological and pathological processes. Here, we show that purified cADPR evoked TRPM2 channel currents in both whole-cell and cell-free single-channel recordings and specific binding of cADPR to the purified NUDT9-H domain of TRPM2 by surface plasmon resonance. Furthermore, by combining computational modeling with electrophysiological recordings, we show that the TRPM2 channels carrying point mutations at H1346, T1347, L1379, S1391, E1409, and L1484 possess distinct sensitivity profiles for ADPR and cADPR. These results clearly indicate cADPR is a bona fide activator at the TRPM2 channel and clearly delineate the structural basis for cADPR binding, which not only lead to a better understanding in the gating mechanism of TRPM2 channel but also shed light on a cADPR-induced RyRs-independent Ca2+ signaling mechanism. : Yu et al. demonstrate that cADPR, which was thought to be an intracellular calcium messenger by modulating function of the ryanodine receptors (RyRs), also activates the TRPM2 channel. Their results suggest that cADPR regulates intracellular calcium release through multiple pathways. Keywords: cADPR, NUDT9-H domain, TRPM2 channel

Dexmedetomidine (Dex) protects different cell types during hypoxia or ischemia-reperfusion injury by inhibiting cell apoptosis. However, the underlying mechanism and its impact on hepatic ischemia reperfusion injury are still not known. In this study, we established a model of oxygen-glucose deprivation/reperfusion (OGD/R) injury in hepatocyte HL7702 cells, and studied the impact of Dex on cell proliferation, apoptosis, and cell cycle during OGD/R. In addition, we explored the role of CCAT1 in this process. We found that Dex increased cell proliferation and inhibited cell apoptosis during OGD/R, in a concentration-dependent manner. Dex partially reversed the OGD-inhibited expression of lncRNA CCAT1. Knockdown of CCAT1 by siRNA inhibited Dex-mediated protection against OGD/R-induced injury and promoted cell apoptosis, caspase-3 expression and cell cycle arrest in the G0/G1 phase, and inhibited cell proliferation and cyclin D1 expression. In contrast, overexpression of CCAT1 by pcDNA3.0-CCAT1 enhanced Dex-mediated protection against OGD/R-induced cell injury. Thus, Dex protects hepatocytes against OGD/R injury by upregulating lncRNA CCAT1. This study suggests a novel role of CCAT1 in ischemia reperfusion injury, and lays the framework for future studies.

Background/Aims: Hepatic ischemia-reperfusion (I/R) injury, which is mainly induced by inflammation and unstable intracellular ions, is a major negative consequence of surgery that compromises hepatic function. However, the exact mechanisms of liver I/R injury have not been determined. Positive crosstalk with the Ca2+/CaMKII pathway is required for complete activation of the TLR4 pathway and inflammation. We previously found that miR-148a, which decreased in abundance with increasing reperfusion time, targeted and repressed the expression of CaMKIIα. In the present study, we examined the role of the miR-148a machinery in I/R-induced Ca2+/CaMKII and TLR4 signaling changes, inflammation, and liver dysfunction in vivo and in vitro. Methods: Liver function was evaluated by serum aminotransferase levels and hematoxylin-eosin (HE) staining. Inflammatory factors were detected by enzyme-linked immunosorbent assay. Gene and protein expression were assessed by RT-PCR and western blot. Small interfering RNA was used to silence target gene expression. HE staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling were used to measure hepatic tissue apoptosis. These assays were performed to identify factors upregulated in hepatic I/R injury and downregulated by miR-148a. Results: We manifested that expression of CaMKIIα and phosphorylation of TAK1 and IRF3 were elevated in hypoxia/reoxygenation (H/R)-treated primary Kupffer cells (KCs) and liver tissue of I/R-treated mice, but these effects were attenuated by treatment with miR-148a mimic and were accompanied by the alleviation of liver dysfunction and hepatocellular apoptosis. Luciferase reporter experiments showed that miR148a suppressed luciferase activity by almost 60%. Moreover, knockdown of CaMKIIα in H/R KCs led to significant deficiencies in p-TAK1, P-IRF3, IL-6, and TNF-α, which was consistent with the effects of miR-148a overexpression. Otherwise, the same trend of activation of TAK1 and IRF3 and inflammatory factors in vitro was observed in the siTAK1 + siIRF3 group compared with the siCaMKIIα group. Conclusion: Taken together, we conclude that miR-148a may mitigate hepatic I/R injury by ameliorating TLR4-mediated inflammation via targeting CaMKIIα in vitro and in vivo.

The east–west trending Tsinling Mountains in central China were uplifted at the end of the Middle Jurassic [176–161 million years ago (Ma)] in Yanshanian, thus effectively and geographically defining the northern climate as cold and dry, and the southern climate as warm and humid. Influenced by paleoenvironmental variation, the karst process shows differences between northern and southern China. Using the systems approach, the authors integrated the geologic history, climate, and hydrological conditions to analyze the causes of the karst differences in northern and southern China, as well as in the Tibetan Plateau. Carbonate rock deposition began in the Mesoproterozoic Era (1,600–1,000 Ma) in north China, and in the Sinian Sub-Era (825–570 Ma) in south China. In north China, the rock formation ended in the Mid-Ordovician (466 Ma), while in South China the deposition continued to the Triassic (250–200 Ma). Tibetan Plateau was deposited in the Late Permian (257–250 Ma). The different depositional environment caused different lithologies: the limestones are largely micritic in the north, but are massive and sparry in the south. The modern karst features were formed mainly in the Tertiary (53–2.6 Ma) and the Quaternary. In the Quaternary, the Tibetan Plateau arose sharply, which formed the monsoon system of East Asia, and loess started to deposit in north China, which partly delayed or prevented karstification in north China, and differentiated the karst features from those in south China. Thus, the karst process in north China is mainly hypogene, while the south is epigene in the Quaternary.

Based on the groundwater development process, and regional economic and social developing history, we divided the spring hydrological process of the Liulin Springs Basin into two periods: pre-1973 and post-1974. In the first period (i.e. 1957–1973), the spring discharge was affected by climate variation alone, and in the second period (i.e. 1974–2009), the spring discharge charge was influenced by both climate variation and human activities. A piecewise analysis strategy was used to differentiate the contribution of anthropogenic activities from climate variation on karst spring discharge depletion in the second period. Then, the ARIMAX model was applied to spring flow time series of the first period to develop a model for the effects of climate variation only. Using this model, we estimated the spring discharge in the second period solely under the influence of climate variation. Based on the water budget, we subtracted observed spring discharge from the estimated spring discharge and acquired the contribution of human activities on spring discharge depletion for the second period. The results of the analysis indicated that the contribution of climate variation to the spring discharge depletion is−0.20 m3/s from 1970s to 2000s. The contribution of anthropogenic activities to the spring flow depletion was −2.56 m3/s in 2000s, which was about 13 times more than that of climate variation. Our analysis further indicates that groundwater exploitation only accounts for 29% of the spring flow depletion due to the effects of human activities. The remaining 71% of the depletion is likely to be caused by other human activities, including dam building, dewatering during coal mining, and deforestation. Copyright © 2012 John Wiley & Sons, Ltd.

Hao, Yonghong, Jiaojuan Zhao, Huamin Li, Bibo Cao, Zhongtang Li, and Tian-Chyi J. Yeh, 2012. Karst Hydrological Processes and Grey System Model. Journal of the American Water Resources Association (JAWRA) 48(4): 656-666. DOI: 10.1111/j.1752-1688.2012.00640.x Abstract: The karst hydrological processes are the response of karst groundwater system to precipitation. This study provided a concept model of karst hydrological processes. The hydraulic response time of spring discharge to precipitation includes the time that precipitation penetrates through the vadose zone, and the subsequent groundwater pressure wave propagates to a spring outlet. Due to heterogeneities in karst aquifers, the hydraulic response time is different in different areas. By using grey system theory, we proposed a karst hydrological model that offers a calculation of hydraulic response time, and a response model of spring discharge to precipitation. Then, we applied the models to the Liulin Springs Basin, China. In the south part of the Liulin Springs Basin, where large fields of carbonate rocks outcrop with intensive karstification, the hydraulic response time is one year. In the north, where the Ordovician karst aquifer is covered by Quaternary loess sediments, the response time is seven years. The grey system GM(1,3) response model of spring discharge to precipitation was applied in consideration of the hydraulic response time. The model calibration showed that the average error was 6.55%, and validation showed that the average error was 12.19%.

GLYX-13, a NMDAR glycine-site partial agonist, was discovered as a promising antidepressant with rapidly acting effects but no ketamine-like side effects. However, the reported synthetic process route had deficiencies of low yield and the use of unfriendly reagents. Here, we report a scaled-up synthesis of GLYX-13 with an overall yield of 30% on the hectogram scale with a column chromatography-free strategy, where the coupling and deprotection reaction conditions were systematically optimized. Meanwhile, the absolute configuration of precursor compound of GLYX-13 was identified by X-ray single crystal diffraction. Finally, the activity of GLYX-13 was verified in the cortical neurons of mice through whole-cell voltage-clamp technique.

To construct a zinc finger protein-artificial transcription factor (ZFP-ATF) specifically bound to hepatitis B virus (HBV) enhancer 1 (Enh1), then detect its expression in HepG2.2.15 cells, observe its effect on cell growth and its role in inhibiting the replication and expression of HBV DNA.The zinc finger protein (ZFP) specifically bound to HBV Enh1 was fused with a Krvppel-associated box (KRAB) transcriptional repression domain to produce an artificial transcription factor (ATF). After relative modification and optimization, the synthetic ATF nucleic acid sequence was inserted into the eukaryotic expression plasmid vector pcDNA3.1⁺ and its sequence was identified. Then pcDNA3.1-ATF was transfected into HepG2.2.15 cells through X-tremeGENE HP. The expression of ATF was detected by confocal microscope. The impact of ATF on cell growth was measured by CCK-8 assay. The inhibitory effect on HBV DNA by ATF was assured by real-time quantitative PCR (qRT-PCR), ELISA and Western blotting at 24, 48 and 72 hours after transfection.The pcDNA3.1-ATF (nls-ZFP-KRAB-FLAG) eukaryotic expression plasmid vector was successfully constructed. ATF was expressed in HepG2.2.15 cells as expected without obvious influence on cell growth. ATF repressed the replication and expression of HBV DNA, especially at 72 hours post-transfection when the inhibition rate was 68%.The synthetic ATF can be expressed in HepG2.2.15 cells normally without toxic effect on the cells and exert an inhibitory effect on the replication and expression of HBV DNA by specifically bound to HBV Enh1.

To explore the impact of transcription factor 21 (TCF21) gene on proliferation, migration and apoptosis in SMMC-7721 hepatocellular carcinoma cell line and the related mechanism.Using Lipofectamine™2000, we stably transfected pcDNA3.1⁺TCF21 and pcDNA3.1⁺ plasmids into SMMC-7721 cells of TCF21 over-expression group and empty vector group, respectively. The untreated cells were set as blank control group. The expression of TCF21 mRNA and protein were investigated by reverse transcription PCR and Western blotting. Cell proliferation ability was detected by MTT assay. Wound healing assay was used to observe cell migration ability. Annexin V-FITC/PI double labeling combined with flow cytometry was applied to determine cell apoptosis rate. Western blotting was performed to examine the expressions of KISS1, P53 and matrix metalloproteinase-9 (MMP-9).TCF21 was over-expressed in TCF21-transfected SMMC-7721 cells. Compared with the control groups, the TCF21 over-expression group showed relatively weakened proliferation ability and significantly inhibited migration ability as well as the increased apoptosis. Western blotting demonstrated that up-regulated TCF21 raised the expressions of KISS1 and p53, and down-regulated MMP-9 level.Tumor-suppressor gene TCF21 could inhibit the proliferation and migration of SMMC-7721 hepatocellular carcinoma cells and promote its apoptosis.

The Liulin karst spring flow has been declining since 1950s, and the main cause to the attenuation is more and more intensive human activities and instructions. By using the Grey system model piecewise, this paper focuses on analyzing the effects of anthropogenic activities to hydrological process of Liulin karst spring statistically. In the first stage (1957–1973), Liulin Springs flow is only affected by climate; in the second stages (1974–1996), Liulin Spring flow is affected by climate and human activities, and not steady; in the third stage (1997–2009), Liulin Springs flow is affected by climate and human activities, and steady. Then through scientifically comparing, we analyzed the effect of human activities to hydrological process of Liulin Spring by using Grey relational analysis and GM model with time lags. The results illustrate that (1) the lag increases from 0 to 4 years; (2) the most important recharge to karst flow is precipitation (3) it is human activities that caused the attenuation of spring flow after 1974, which means that human activities have significantly negative influences on the supplement of precipitation to Liulin Spring.