Vegetation-fire feedbacks increase subtropical wildfire risk in scrubland and reduce it in forests.

Climate dictates wildfire activity around the world. But East and Southeast Asia are an apparent exception as fire-activity variation there is unrelated to climatic variables. In subtropical China, fire activity decreased by 80% between 2003 and 2020 amid increased fire risks globally. Here, we assessed the fire regime, vegetation structure, fuel flammability and their interactions across subtropical Hubei, China. We show that tree basal area (TBA) and fuel flammability explained 60% of fire-frequency variance. Fire frequency and fuel flammability, in turn, explained 90% of TBA variance. These results reveal a novel system of scrubland-forest stabilized by vegetation-fire feedbacks. Frequent fires promote the persistence of derelict scrubland through positive vegetation-fire feedbacks; in forest, vegetation-fire feedbacks are negative and suppress fire. Thus, we attribute the decrease in wildfire activity to reforestation programs that concurrently increase forest coverage and foster negative vegetation-fire feedbacks that suppress wildfire.

Efficacy of a formalin-inactivated Lactococcus garvieae vaccine in farmed grey mullet (Mugil cephalus).

Lactococcosis [Lactococcus garvieae (LG)] is one of the most prevalent bacterial diseases affecting grey mullet (Mugil cephalus) aquaculture. Therefore, the present research evaluated the efficacy of formalin-killed LG vaccine with an oil-based adjuvant in grey mullet under laboratory and field trials. The laboratory evaluation for LG vaccine and its cross-protection upon challenge in grey mullet found that single-dose immunization of formalin-killed LG with adjuvant resulted in 91.4% and 100% relative per cent survival (RPS) when challenged with homologous and heterologous strains. The levels of specific antibody titre and lysozyme activity increased significantly in the vaccinated group. Immune gene expression at 24 hr after challenge showed an increase in levels of pro-inflammatory and anti-inflammatory cytokines. A parallel field trial experiment was conducted to investigate the long-term effectiveness of the LG vaccine. Results demonstrated that at one month and three months post-immunization with heterologous strain, 100% RPS was recorded in the vaccinated group. The findings suggested that the formalin-inactivated LG vaccine strain (S3) protected grey mullet against LG infection for a period of three months.

Current knowledge of nocardiosis in teleost fish.

Nocardia sp. is the causative agent of nocardiosis, a lethal granulomatous disease of the skin, muscle, and various inner tissues affecting various teleost and shellfish. Four species of Nocardia have been isolated from diseased fish and shellfish, namely Nocardia asteroides, Nocardia seriolae, Nocardia salmonicida and Nocardia crassostreae. Therefore, in fish aquaculture, nocardiosis has caused severe economic losses, especially in the Asian region. Considerable research has been performed, since the first report of identified Nocardia sp. in fish, to characterize Nocardia sp. and identify rapid detection techniques, immune response against infection and prophylactic approaches. In this review, the current state of knowledge about nocardiosis in fish has been presented, including the pathogenesis, diagnosis, host immune response and vaccine development.

Effect of karst rocky desertification on soil fungal communities in Southwest China.

Karst mountainous ecosystems are associated with karst rocky desertification (KRD), which can greatly impact soil structure and function. Despite the importance of soil microbes as a major factor maintaining ecosystem stability, we know little about the effect on soil fungal communities of KRD in karst regions. We investigated this relationship across a gradient of KRD soils from Guizhou, China by polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE). Fungal diversity indices (Shannon-Wiener, richness, and evenness) significantly differed (P < 0.05) based on KRD severity, being lowest in moderately affected areas. Cluster analysis showed that the five sites examined clustered into two main groups according to KRD grade (high and low). Moreover, a homology search using sequences recovered from PCR-DGGE bands showed that the dominant fungi in each community varied remarkably, and included Aspergillus, Aphanoascus, Blastomyces, Fusarium, Glomus, Geomyces, Gibberella, Mortierella, Tetracladium, and Tumularia species, and an unclassified group. In conclusion, these findings demonstrate that KRD has a significant impact on soil fungal communities.

Genetic characteristics of Streptococcus dysgalactiae isolated from cage cultured cobia, Rachycentron canadum (L.).

Disease outbreaks occurred during 2007-2013 in Taiwan with 2.5-10% mortality among the cage cultured cobia, Rachycentron canadum (L.), characterized by the presence of polyserositis, pericarditis and peritonitis. The micro-organisms isolated from internal organs were Gram-positive cocci. The isolates were confirmed as Streptococcus dysgalactiae by a polymerase chain reaction assay that yielded the expected specific 259 bp amplicon. Additionally, partial sequence of the 16S-23S rDNA intergenic spacer region of the GCS strain isolates from fish was also compared and produced 100% sequence identity with S. dysgalactiae (GenBank accession number AB252398). The genetic characterization was then determined by pulsed-field gel electrophoresis (PFGE) analysis. Based on PFGE, the Apa I or Sma I digestion patterns of chromosomal DNA of these isolates were grouped into three main clusters. Taiwanese strains were divided into two clusters, and the tet(M) gene was detected in cluster 1 (pulsotypes: A1-A2 and S1-S3), but not in cluster 2 strains (pulsotypes: A3-A4 and S4-S5). Three Japanese strains from amberjack, Seriola dumerili (Risso), were grouped into cluster 3 (pulsotypes: A5-A7 and S6-S8) and displayed no mortality to cobia in the challenge experiment. Conversely, Taiwanese strains from cobia and snubnose pompano, Trachinotus blochii (L.), displayed a mortality rate of 50-87.5% in cobia.

Genetic diversity of Ceratoides arborescens, a species endemic to China, detected by inter-simple sequence repeat (ISSR).

In order to investigate genetic diversity and population structure of Ceratoides arborescens, six populations were selected from different steppe types in Inner Mongolia grasslands of China. Inter-simple sequence repeat (ISSR) markers were used to assess the genetic diversity within and among natural populations of C. arborescens. Thirteen ISSR primers generated 154 discernible DNA bands, of which 151 (98.05%) were polymorphic. High genetic diversity was detected at the species level [percentage of polymorphic loci (PPB) = 98.05%; H = 0.2984; I = 0.4557], whereas, relatively low genetic diversity existed within populations (PPB = 80.62%; H = 0.2675; I = 0.4031). Analysis of molecular variance showed that variation existed mainly within populations (73.25%) rather than among populations (26.75%), which was in line with the high level of gene flow (Nm = 4.3332). The Mantel test found no significant correlation between genetic distance and geographic distance (r = 0.7522, P < 0.05). Six populations were clustered into two main groups, a desert steppe group and a typical steppe group.

Hypoxia inducible factor 2α/insulin-like growth factor receptor signal loop supports the proliferation and Oct-4 maintenance of mouse germline stem cells.

Hypoxia inducible factor 2α (HIF-2α) is critical for primordial germ cell (PGC) survival as knockout of HIF-2α (HIF-2α(-/-)) decreases both expression of Oct-4 and PGC number in genital ridge. Hypoxia is known to stabilize HIF-2α protein from proteasomal degradation. However, little is known about the hypoxia-associated endocrinal signaling in HIF-2α expression. The current work demonstrates a role for an endocrine insulin-like growth factor-I receptor (IGF-IR)-PI3K/Akt-mTOR-HIF-2α regulatory loop in the proliferation and Oct-4 maintenance of PGC-like alkaline phosphatase positive mouse germline stem cells (AP(+)GSCs). We found that hypoxia greatly increased the cell proliferation and the levels of nuclear Oct-4/HIF-2α protein of AP(+)GSCs. The hypoxic-AP(+)GSCs presented stronger stemness ability for germ cell differentiation than normoxic, with expressions of c-KIT (differentiation germ cell marker), VASA (differentiation germ cell marker) and SCP3 (meiotic marker) using a renal capsule transplantation assay. Meanwhile, hypoxia significantly increased the expression levels of secreted-IGF-I and IGF-IR. The IGF-I dose dependently increased the HIF-2α expression levels in AP(+)GSCs; and, the inhibition of IGF-IR by RNA interference (shIGF-IR) or LY294002 (PI3K inhibitor)/Rapamycin (mTOR inhibitor) effectively suppressed the IGF-I- and/or hypoxia-induced HIF-2α and Oct-4 expression, suggesting that the IGF-IR and its downstream Akt/mTOR signaling are involved in the IGF-I/hypoxia effects. Additionally, knockdown of HIF-2α dramatically suppressed Oct-4 and IGF-IR protein levels in AP(+)GSC cells. In conclusion, the present study demonstrates a regulatory loop of IGF-IR-PI3K/Akt-mTOR-HIF-2α in proliferation and Oct-4 maintenance of PGC-like AP(+)GSCs under hypoxia. This finding provides insights into the niche endocrinology underlying early germ cell development.

Reduction of galectin-3 expression reduces pituitary tumor cell progression.

We investigated the role of galectin 3 in the development and progression of pituitary tumors. We used RNA interference to depress Gal-3 gene expression; MTT and flow cytometry were applied to detect changes in cell proliferation and apoptosis levels in pituitary tumor cells. Expression of apoptosis-associated genes, Bcl-2 and Baxk was analyzed by Western blot. Inhibition of Gal-3 gene expression by RNA interference decreased GH3 cell proliferation and increased cell apoptosis; the expression levels of Gal-3 protein significantly decreased, along with those of Bcl-2, although Bax levels did not change. We conclude that Gal-3 has an important role in pituitary tumor cell proliferation and progression; and it may be useful as a target for the treatment of aggressive pituitary tumors.

Development of a sensitive and specific LAMP PCR assay for detection of fish pathogen Lactococcus garvieae.

Based on use of a loop-mediated isothermal amplification (LAMP) identification protocol, this study attempted to detect Lactococcus garvieae in fish by using primer sets designed from an L. garvieae alpha/beta fold family hydrolase gene. Reaction time and temperatures were optimized for 60 min at 60°C with the resulting amplicons visualized by adding SYBR Green I to the reaction tube. The assay specificity was assessed using 45 different bacterial strains. Positive results were observed in all 30 L. garvieae isolates from various aquatic animals. No false-positive results were observed in 15 non-L. garvieae strains. The detection limit of the LAMP assay was 10-fold more sensitive than the conventional polymerase chain reaction (PCR) targeting 16S rDNA when using purified L. garvieae DNA. The detection limit of the LAMP assay was approximately 300 colony-forming units (CFU) using crude bacterial lysates, 100-fold more sensitive than PCR. Furthermore, L. garvieae in spleen, kidney and brain of experimentally challenged tilapia and grey mullet were detected using this optimized LAMP assay. Results of this study demonstrate the effectiveness of LAMP in providing a rapid yet simple test for detecting L. garvieae in fish.

White spot syndrome virus epizootic in cultured Pacific white shrimp Litopenaeus vannamei (Boone) in Taiwan.

White spot syndrome virus (WSSV) has caused significant losses in shrimp farms worldwide. Between 2004 and 2006, Pacific white shrimp Litopenaeus vannamei (Boone) were collected from 220 farms in Taiwan to determine the prevalence and impact of WSSV infection on the shrimp farm industry. Polymerase chain reaction (PCR) analysis detected WSSV in shrimp from 26% of farms. Juvenile shrimp farms had the highest infection levels (38%; 19/50 farms) and brooder shrimp farms had the lowest (5%; one of 20 farms). The average extent of infection at each farm was as follows for WSSV-positive farms: post-larvae farms, 71%; juvenile farms, 61%; subadult farms, 62%; adult farms, 49%; and brooder farms, 40%. Characteristic white spots, hypertrophied nuclei and basophilic viral inclusion bodies were found in the epithelia of gills and tail fans, appendages, cephalothorax and hepatopancreas, and virions of WSSV were observed. Of shrimp that had WSSV lesions, 100% had lesions on the cephalothorax, 96% in gills and tail fans, 91% on appendages and 17% in the hepatopancreas. WSSV was also detected in copepoda and crustaceans from the shrimp farms. Sequence comparison using the pms146 gene fragment of WSSV showed that isolates from the farms had 99.7-100% nucleotide sequence identity with four strains in the GenBank database--China (AF332093), Taiwan (AF440570 and U50923) and Thailand (AF369029). This is the first broad study of WSSV infection in L. vannamei in Taiwan.

EMG-guided salpingopharyngeus Botox injection for palatal myoclonus.

Palatal myoclonus (PM) is a rare neurological disorder characterized by involuntary movements of the soft palate musculature causing objective clicking tinnitus. Two forms are recognized as distinct clinical entities, with poorly understood pathogenesis: essential and symptomatic PM. The intrusive nature of the tinnitus prompts patients to seek medical advice. Conventional medical treatments with anxiolytics, antidepressants, and anticonvulsants have limited efficacy in these patients. In this case report, electromyography-guided injection of botulinum toxin type A (Botox; Allergan, Irvine, CA, USA) to the involved salpingopharyngeus and tensor veli palatini yielded satisfactory results with minimal temporary adverse effects.

Auditory efferent dysfunction in normal-hearing chronic idiopathic tinnitus.

OBJECTIVE: To investigate the function of the auditory efferent system in patients with chronic idiopathic tinnitus, but normal pure-tone audiograms. METHODS: We studied 15 subjects with normal hearing that had experienced either unilateral or bilateral persistent tinnitus for at least 3 months. The ears of the 15 subjects were classified into tinnitus-positive-ear (TPE) and tinnitus-negative-ear (TNE) groups. The control-ear group (CE) comprised the ears of 15 subjects with normal hearing and no tinnitus. We measured different types of otoacoustic emissions (OAEs), including spontaneous (SOAEs), transient evoked (TEOAEs), and distortion product (DPOAEs). We also analyzed contralateral suppression of OAEs and auditory brainstem responses (ABRs). Data were compared among TPE, TNE, and CE groups. RESULTS: The data associated with cochlear mechanics, including the prevalence of SOAEs, the number of SOAE peaks, and the overall TEOAE responses in the absence of a contralateral stimulus, were not significantly different among the TPE, TNE, and CE groups. In the TPE group, contralateral stimuli failed to significantly suppress overall TEOAEs, and contralateral suppression of DPOAEs was significantly reduced over a limited frequency range. Furthermore, the TPE group showed prolonged latencies in waves III and V of ABRs. CONCLUSION: This study demonstrated that abnormal contralateral suppression of OAEs and ABRs indicated a dysfunction in the ipsilateral efferent medial olivocochlear system; this might play a role in normal-hearing tinnitus.

Safety and efficacy of two-step peginterferon α-2a treatment in patients of chronic hepatitis B with acute exacerbation.

The focus of this study was to evaluate the safety and efficacy of sequential peginterferon α-2a (Pegasys) therapy for chronic hepatitis B with acute exacerbation [ALT > 10 × upper limit of normal (ULN), bilirubin <2.0 mg/dL]. Four groups of patients categorized by HBeAg status and treatment regimens were studied since May 2007. Nineteen HBeAg-positive patients (Group 1) had received entecavir  pretreatment  (when ALT > 10 × ULN) plus Pegasys (180 µg/kg/week, when ALT was 5-10 × ULN) for 24 weeks. Thirteen HBeAg-negative patients (Group 2) had the same protocol for 48 weeks. In both groups, entecavir was then discontinued 14 days after the initiation of Pegasys. The results were compared, respectively, to 35 HBeAg-positive patients (Group 3) and 24 HBeAg-negative patients (Group 4), all with ALT > 5 × ULN, under continual entecavir monotherapy. The ALT levels of patients in Group 1 and 2 who had received entecavir pretreatment for a duration of 19.63 ± 3.34 days were below four times of ULN following 4 weeks of Pegasys treatment. At week 96, the rates of sustained virological response were 69.2% (9/13) and 80% (8/10), and the relapse rates were 23.1% (3/13) and 11.2% (1/9) for HBeAg-positive and HBeAg-negative patients with two-step Pegasys treatment, respectively. The HBeAg seroconversion rates were 46.2% in Group 1, and 42.1% in Group 3; HBsAg loss rates were 15.4% (2/13) in Group 1, and 30% (3/10) in Group 2, whereas none achieved HBsAg loss with entecavir monotherapy (Group 3 and 4). The two-step Pegasys treatment offers an alternative, other than the nucleos(t)ides, for treating chronic hepatitis B with acute exacerbation and provides a safe, efficacious, short-term and finite strategy.

Development of a multiplex polymerase chain reaction to detect five common Gram-negative bacteria of aquatic animals.

A multiplex polymerase chain reaction (m-PCR) technique was developed as a rapid and accurate diagnostic tool for identifying five major Gram-negative bacilli -Vibrio vulnificus, V. parahaemolyticus, Aeromonas hydrophila, Chryseobacterium meningosepticum and Edwardsiella tarda- that cause major diseases in cultured aquatic animals in Taiwan. The expected amplicons for V. vulnificus, V. parahaemolyticus, A. hydrophila, C. meningosepticum and E. tarda were 410, 368, 685, 180 and 230bp, respectively. The assay was shown to be specific for the target pathogens. The sensitivities of detection were estimated to be 20.5fg∼200pg of genomic DNA or 10(2) ∼10(4) colony-forming units (cfu) of bacterial isolates when adopted as PCR templates. The m-PCR was capable of simultaneously amplifying target fragments from bacterial genome DNA mixed with the DNA extracted from viscera and tissues taken from fish without affecting the performance of the method.

Koi herpesvirus epizootic in cultured carp and koi, Cyprinus carpio L., in Taiwan.

Koi herpesvirus (KHV) poses a significant threat to cultured koi and common carp, both Cyprinus carpio L. Since the first reported case in Israel in 1998, KHV has rapidly spread worldwide. This study investigates the spread of KHV to Taiwan by collecting 49 cases of suspected common carp and koi infections from 2003 to 2005 for analysis. Clinical signs included lethargy, anorexia, increased respiratory movements and uncoordinated swimming. Hyperaemia, haemorrhage on body surface and necrotic gill filaments were recorded. Gill epithelial hyperplasia, necrosis and eosinophilic intranuclear inclusion bodies were observed by histological examination, while virions were detected using transmission electron microscopy. By detecting the presence of the KHV thymidine kinase (TK) gene and the KHV 9/5 gene using polymerase chain reaction (PCR), 37 cases were identified as KHV-positive, and the cumulative mortality of infected fish was 70-100%. Positive cases showed identical sequences for the genes analysed, implying that they were of the same origin. For the KHV 9/5 gene sequence, these cases exhibited 100% identity with the Japanese strain (TUMST1, accession number AP008984) and 99% identity with the Israeli (KHV-I, DQ177346) and US (KHV-U, DQ657948) strains. Additionally, a loop-mediated isothermal amplification (LAMP) assay was performed and found to be more sensitive than PCR tests, suggesting its potential use as a rapid diagnostic method for KHV. This is the first epidemiological study of KHV infection in cultured common carp and koi in Taiwan.

The influence of personality traits on nurses' job satisfaction in Taiwan.

AIM: The purpose of this study was to empirically investigate a set of personality traits based on core self-evaluations in relation to job satisfaction among nurses in Taiwan, and to identity important personality characteristics in contributing to nurses' satisfactory attitudes towards work. METHOD: A cross-sectional survey research was conducted with self-reported questionnaires. The participants were 314 nurses (89% response rate) in two regional hospitals in Taiwan. Among them, 99% were female (n=311), 54.5% (n=171) ranging between 21 and 30 years old, 37.9% (n=119) ranging between 31 and 40 years old, 44.6% (n=140) are registered nurse, 29.3% (n=92) are licensed vocational nurse and 52.9% were married (n=166). In addition, ethical research requirements for conducting research on human subjects were fulfilled for all of the participants in the study. ANALYSIS: Data were analysed using SPSS version 14.0 software (SPSS Inc., Chicago, IL, USA). A number of data analysis procedures were conducted including descriptive statistics, Pearson product-moment correlation and multiple regression analysis. The selection of the predictor variables was performed with the level of significance P<0.05 and the adjusted explained variance was used to examine statistical explanatory power. RESULTS: Personality traits of optimism, self-esteem, proactive personality, self-efficacy and negative affectivity significantly contributed to job satisfaction. CONCLUSIONS: The findings of this study confirmed the importance of taking personality variables into consideration during the process of evaluating job satisfaction. Job satisfaction should not only be related to extrinsic factors but also associated with individual differences of dispositional tendency. Nurses with positive evaluation and expectation towards self and others tend to report higher job satisfaction. The results of this study are consistent with previous studies. The implications for nursing managements are discussed.

Nocardia seriolae infection in the three striped tigerfish, Terapon jarbua (Forsskål).

An epizootic in pond cultured three striped tigerfish, Terapon jarbua, in Taiwan was caused by Nocardia seriolae. Diseased fish first showed clinical signs and mortalities in February and March 2003. The cumulative mortality within 2 months was 2.4% (1200 of 50 000) and affected fish were 7 months old with total lengths from 18 to 25 cm. Most affected fish were pale and lethargic with haemorrhages and ulcers on the skin. The most significant gross pathological changes were varying degrees of ascites and enlargement of the spleen, kidney and liver. Obvious white nodules, varying in size, were found in these organs. Bacteria were either coccal or filamentous in appearance, with bead-like forms. Isolates from diseased fish were characterized using the API ZYM (Analytical profile index; Bio Mérieux, France) systems and conventional tests and identified as Nocardia sp. The isolate was designated NS127 and was confirmed as N. seriolae by a polymerase chain reaction assay that gave the expected specific 432 bp amplicon. In addition, its 16S rDNA sequence gave 100% sequence identity with N. seriolae. A partial sequence of the 16S rRNA gene, heat shock protein gene and RNA polymerase gene (rpo B) of NS127 and the type strain of N. seriolae BCRC 13745 formed a monophyletic clade with a high sequence similarity and bootstrap value of 99.9%. White nodules induced in experimental fish were similar to naturally infected cases and N. seriolae was re-isolated on brain heart infusion agar. This is the first report of N. seriolae-infection in three striped tigerfish in aquaculture.

A new delineation method research of the clinical target volume for pancreatic cancer adjuvant radiotherapy.

PURPOSE: The purpose of this work was to establish a map model of the local recurrence location after pancreatic cancer resection and to generate a new delineation method of clinical target volume, with the aim to effectively improve the adjuvant radiotherapeutic gain ratio. METHODS AND MATERIALS: The clinical and imaging data of 48 patients with resected pancreatic head cancer and pancreatic body cancer with local recurrences were collected. Local recurrences were all plotted with reference to the geometric centre of the local recurrent foci. Based on the coordinates of the local recurrences with respect to the celiac artery or the superior mesenteric artery, a three-dimensional local recurrence map model was established on the computed tomography image. The adjuvant radiation clinical target volumes encompassing 90% of all local failures and encompassing 90% of postoperative pancreatic head cancer local failures were created respectively. This new delineation method and RTOG 0848 protocol were applied in five simulated cases, then corresponding types of target volumes and plans were generated for comparison. RESULTS: The clinical target volume encompassing 90% of all local failures was generated by expanding the combined celiac artery and superior mesenteric artery contour by 1.4cm superior, 1.9cm inferior, 2.6cm left-lateral, 3.1cm right-lateral, 1.9cm anterior and 1.6cm posterior. The corresponding expansions of clinical target volume encompassing 90% of postoperative pancreatic head cancer local failures were 1.4cm, 1.4cm, 2.1cm, 3.1cm, 1.6cm and 2.0cm. The volumes of "new" target PTV-90_edited, PTV-90_H_edited, and the standard target PTV_edited were 217.64±58.67 cm3, 207.78±50.94 cm3 and 320.72±50.94 cm3 in simulated cases. Comparison showed that the "new" target volumes were much smaller than the standard volumes per RTOG 0848 protocol, and the dose received by organs at risk was also lower in the "new" plans. CONCLUSIONS: A majority of postoperative local recurrences in patients with pancreatic head and body cancer are contained within a smaller region surrounding the celiac artery and superior mesenteric artery. The "new" volumes targeting high risk local failures may allow dose escalation and enhanced local control while minimizing radiation-related toxicity.

Cisplatin suppresses tumor proliferation by inhibiting autophagy in ovarian cancer via long non-coding RNA RP11-135L22.1.

OBJECTIVE: To observe the effect of cisplatin-induced autophagy in human ovarian cancer cell lines and explore the correlation between RP11-135L22.1 with cisplatin-induced autophagy. MATERIALS AND METHODS: Genome-wide expression profile and chemotherapy sensitivity data of ovarian cancer were downloaded from TCGA database. It was found that the expression level of lncRNA RP11-135L22.1 differed between chemotherapy-sensitive group and insensitive group. Besides, RP11-135L22.1 expression levels were detected in 64 ovarian cancer tissues and 30 normal tissues by qRT-PCR. Relationship between RP11-135L22.1 expression levels in 64 ovarian cancer tissues and their clinicopathological characteristics were analyzed by x2-test. Cell viability was detected by CCK8 assay. Cell apoptosis and cell cycle were accessed by flow cytometry. HO8910 cells were selected for transfection of pcDNA-RP11-135L22.1, and qRT-PCR was used to evaluate RP11-135L22.1 expression in cisplatin-treated HO8910 cells. Western blot was performed to analyze the expression changes of autophagy-related proteins. RESULTS: Genome-wide expression profile of chemotherapy-sensitive and -insensitive patients with ovarian cancer from TCGA database was analyzed by edger package. It was found that RP11-135L22.1 level in chemotherapy-sensitive group was significantly lower than that of insensitive group. QRT-PCR results confirmed that RP11-135L22.1 was lowly expressed in ovarian cancer. The overall survival of patients was positively correlated with the expression of RP11-135L22.1. Furthermore, RP11-135L22.1 was associated with FIGO stage and tumor size. Flow cytometry showed that cisplatin could induce apoptosis and arrest cell cycle in ovarian cancer cells lines. CCK8 assay showed that cisplatin decreased viability of ovarian cancer cells. For in vitro study, HO8910 cells were cultured with medium containing different concentrations of cisplatin or treated with cisplatin for different times. The results revealed that RP11-135L22.1 expression was negatively correlated with the treating time and dose of cisplatin. Western blot showed that cisplatin induced autophagy in ovarian cancer cells in a time- and dose-dependent manner. Cisplatin combined with RP11-135L22.1 can reduce autophagy, increase the apoptosis and inhibit its activity of ovarian cancer cells to a certain extent. CONCLUSIONS: Cisplatin can induce autophagy in HO8910 ovarian cancer cells. After overexpression of RP11-135L22.1, it inhibited cisplatin-induced autophagy, thus enhancing the effect of cisplatin on ovarian cancer cells.

Ibrutinib presents antitumor activity in skin cancer and induces autophagy.

OBJECTIVE: Skin cancer is one of the most common malignancies in dermatology. Patient compliance and prognosis of skin cancer are poor. Ibrutinib, a Bruton's Tyrosine Kinase (BTK) inhibitor, is a new anticancer drug used to treat many cancers. Therefore, we aimed to explore the role of ibrutinib in the treatment of skin cancer. MATERIALS AND METHODS: Cell Counting Kit-8 (CCK8) and plate cloning assay were used to detect cell proliferation. Apoptosis was determined by flow cytometry. Western blotting analysis was used to analyze the expression of key proteins that regulated autophagy. Proliferation and apoptosis of skin cancer cells and induction of autophagy induced by ibrutinib were evaluated. RESULTS: CCK8 plate cloning assays showed that ibrutinib can gradually inhibit the skin cancer cell proliferation as the treatment time and dose increased. Results of flow cytometry showed that apoptosis in skin cancer cells were induced after ibrutinib treatment. Western blot showed that autophagy in skin cancer cells was found induced by ibrutinib and also related to the time and concentration of ibrutinib treatment. Combination treatment of ibrutinib and 3MA for skin cancer cells can significantly increase apoptosis. CONCLUSIONS: Ibrutinib has anti-tumor activity in skin cancer and can induce autophagy. Binding to autophagy inhibitors can promote ibrutinib's anti-skin cancer activity. Our experimental results provided new ideas for developing skin cancer drugs.