Dexmedetomidine suppressed the biological behavior of RAW264.7 cells treated with LPS by down-regulating HOTAIR.

Background: Previous studies have revealed dexmedetomidine have potential protective effects on vital organs by inhibiting the release of inflammatory cytokines. To investigate the effects of dexmedetomidine on sepsis, especially in the initial inflammatory stage of sepsis. RAW264.7 cells were used as the cell model in this study to elucidate the underlying mechanisms. Methods: In this study, we conducted several assays to investigate the mechanisms of dexmedetomidine and HOTAIR in sepsis. Cell viability was assessed using the CCK-8 kit, while inflammation responses were measured using ELISA for IL-1ß, IL-6, and TNF-α. Additionally, we employed qPCR, MeRIP, and RIP to further explore the underlying mechanisms. Results: Our findings indicate that dexmedetomidine treatment enhanced cell viability and reduced the production of inflammatory cytokines in LPS-treated RAW264.7 cells. Furthermore, we observed that the expression of HOTAIR was increased in LPS-treated RAW264.7 cells, which was then decreased upon dexmedetomidine pre-treatment. Further investigation demonstrated that HOTAIR could counteract the beneficial effects of dexmedetomidine on cell viability and cytokine production. Interestingly, we discovered that YTHDF1 targeted HOTAIR and was upregulated in LPS-treated RAW264.7 cells, but reduced in dexmedetomidine treatment. We also found that YTHDF1 increased HOTAIR and HOTAIR m6A levels. Conclusions: Collectively, our results suggest that dexmedetomidine downregulates HOTAIR and YTHDF1 expression, which in turn inhibits the biological behavior of LPS-treated RAW264.7 cells. This finding has potential implications for the prevention and treatment of sepsis-induced kidney injury.

The cotton MYB33 gene is a hub gene regulating the trade-off between plant growth and defense in Verticillium dahliae infection.

INTRODUCTION: Sessile plants engage in trade-offs between growth and defense capacity in response to fluctuating environmental cues. MYB is an important transcription factor that plays many important roles in controlling plant growth and defense. However, the mechanism behind how it keeps a balance between these two physiological processes is still largely unknown. OBJECTIVES: Our work focuses on the dissection of the molecular mechanism by which GhMYB33 regulates plant growth and defense. METHODS: The CRISPR/Cas9 technique was used to generate mutants for deciphering GhMYB33 functions. Yeast two-hybrid, luciferase complementary imaging, and co-immunoprecipitation assays were used to prove that proteins interact with each other. We used the electrophoretic mobility shift assay, yeast one-hybrid, and luciferase activity assays to analyze GhMYB33 acting as a promoter. A ß-glucuronidase fusion reporter and 5' RNA ligase mediated amplification of cDNA ends analysis showed that ghr-miR319c directedly cleaved the GhMYB33 mRNA. RESULTS: Overexpressing miR319c-resistant GhMYB33 (rGhMYB33) promoted plant growth, accompanied by a significant decline in resistance against Verticillium dahliae. Conversely, its knockout mutant, ghmyb33, demonstrated growth restriction and concomitant augmentation of V. dahliae resistance. GhMYB33 was found to couple with the DELLA protein GhGAI1 and bind to the specific cis-elements of GhSPL9 and GhDFR1 promoters, thereby modulating internode elongation and plant resistance in V. dahliae infection. The ghr-miR319c was discovered to target and suppress GhMYB33 expression. The overexpression of ghr-miR319c led to enhanced plant resistance and a simultaneous reduction in plant height. CONCLUSION: Our findings demonstrate that GhMYB33 encodes a hub protein and controls the expression of GhSPL9 and GhDFR1, implicating a pivotal role for the miR319c-MYB33 module to regulate the trade-offs between plant growth and defense.

Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer.

Exosomes are messengers for intercellular communication and signal transduction. Circular RNA (circRNA) abnormal expression and regulation are involved in the occurrence and development of a variety of tumors. In the present study, exosomes in the serum of five patients with non-small cell lung cancer (NSCLC) were isolated before and after EGFR-TKIs resistance, and the circRNA expression profile was screened using a circRNA microarray. The effects of the exosome circRNA_102481 on cell proliferation and apoptosis were analyzed. The interaction between miR-30a-5p and circRNA_102481 or ROR1 was predicted by starBase software, and was confirmed by RNA pull-down and dual-luciferase reporter assays. The results showed that exosomes containing circRNA_102481 were significantly up-regulated in NSCLC with EGFR-TKIs resistance (p<0.05), and that circRNA_102481 was mainly secreted by EGFR-TKIs resistance cell via exosomes (p<0.05). Both circRNA_102481 silencing and si-circRNA_102481 transported by exosomes could inhibit EGFR-TKIs resistance cell proliferation and promote cell apoptosis and circRNA_102481 overexpression could promote EGFR-TKIs sensitive cell proliferation and inhibit cell apoptosis in vitro (p<0.05). CircRNA_102481 served as a miR-30a-5p sponge to regulate ROR1 expression (p<0.05). Furthermore, the expression of circRNA_102481 in exosomes was associated with TNM stage, tumor differentiation status, brain metastasis, and PFS and OS duration. Therefore, it was concluded that tumor-derived exosomal circRNA_ 102481 could contribute to EGFR-TKIs resistance via the microRNA-30a-5p/ROR1 axis in NSCLC. Exosomal circRNA_102481 may serve as a novel diagnostic biomarker and a therapeutic target for EGFR-TKIs resistance in NSCLC.

Assessment of control strategies against Clonorchis sinensis infection based on a multi-group dynamic transmission model.

Clonorchiasis is one of the most important food-borne trematodiases affecting millions of people. Strategies were recommended by different organizations and control programmes were implemented but mostly in short-time periods. It's important to assess the long-term benefits and sustainability of possible control strategies on morbidity control of the disease. We developed a multi-group transmission model to describe the dynamics of C. sinensis transmission among different groups of people with different raw-fish-consumption behaviors, based on which, a full model with interventions was proposed and three common control measures (i.e., preventive chemotherapy, information, education, and communication (IEC) and environmental modification) and their possible combinations were considered. Under a typical setting of C. sinensis transmission, we simulated interventions according to different strategies and with a series of values of intervention parameters. We found that combinations of measures were much beneficial than those singly applied; higher coverages of measures had better effects; and strategies targeted on whole population performed better than that on at-risk population with raw-fish-consumption behaviors. The strategy recommended by the government of Guangdong Province, China shows good and sustainable effects, under which, the infection control (with human prevalence <5%) could be achieved within 7.84 years (95% CI: 5.78-12.16 years) in our study setting (with original observed prevalence 33.67%). Several sustainable strategies were provided, which could lead to infection control within 10 years. This study makes the effort to quantitatively assess the long-term effects of possible control strategies against C. sinensis infection under a typical transmission setting, with application of a multi-group dynamic transmission model. The proposed model is easily facilitated with other transmission settings and the simulation outputs provide useful information to support the decision-making of control strategies on clonorchiasis.

[Molluscicidal effects of different formulations of niclosamide ethanolamine salt in marshlands].

OBJECTIVE: To evaluate the molluscicidal effects of different formulations of niclosamide ethanolamine salt in marshlands. METHODS: The molluscicidal effects of spraying with 25% suspension concentrate of niclosamide ethanolamine salt (25% SCN) and 50% wettable powder formulation of niclosamide ethanolamine salt (50% WPN), and dusting with 4% niclosamide ethanolamine salt dustable powder (4% DP) for controlling Oncomelania hupensis snails were investigated and compared in the fields, and the cost-effectiveness was analyzed. RESULTS: The corrected mortalities and the reduced rates of density of snails were 54.37%, 91.70%, 92.76%, 79.50%, and 59.55%, 95.93%, 97.63%, 94.15%, respectively, on 3, 7, 15, 30 d after spaying with 25% SCN, those on 3, 7, 15, 30 d after dusting with 4% DP were 59.10%, 91.83%, 95.56%, 93.34% and 65.03%, 94.93%, 97.61%, 97.28%, respectively; and those on 3, 7, 15, 30 d after spraying with 50% WPN were 76.29%, 91.68%, 93.12%, 81.59% and 81.24%, 97.02%, 97.84%, 95.27%, respectively. The cost of spraying with 25% SCN was 0.21 Yuan/m2, that of dusting with 4% DP was 0.39 Yuan/m2, and that of spraying with 50% WPN was 0.23 Yuan/m2 for snail control in the marshland. The cost of reduced one percentage of the corrected mortalities and the density of snails in controlling snails by 25% SCN, 4% DP and 50% WPN on 15 d were 22.68, 40.63, 25.17 Yuan and 21.54, 39.78, 23.95 Yuan, respectively. CONCLUSIONS: The three different formulations of niclosamide are reliable and effective for snail control in marshlands. There are some differences among the different molluscicides in start time, pharmacodynamic characteristics, spraying methods in the field, cost of snail control, and influencing factors. Therefore, we need reasonably select the suitable molluscicides according to the environmental characteristics and working condition.

[Molluscicidal effect of niclosamide ethanolamine salt powder - granula against Oncomelania hupensis].

OBJECTIVE: To evaluate the molluscicidal effect of niclosamide ethanolamine salt powder-granula (PG) against Oncomelania hupensis. METHODS: The molluscicidal experiment was carried out by the dusting method with niclosamide ethanolamine salt 4% PG. The experiments were respectively done in the laboratory and the tidal flats wetlands. At the same time, the niclosamide ethanolamine salt 4% dustable powder (DP) was as the control group. The single blind method was used for the quality control. The corrected mortality and the median lethal concentration (LC50) were compared between PG and DP in the molluscicidal experiment of the laboratory. The corrected mortality and the reduced rate of snails'density were compared between PG and DP in the tidal flats wetlands. RESULTS: The mortality rates of the snails were 96.67% and 100% respectively on 1 d after dusting 4.0 g/m2 of 4% PG and 2.0 g/m2 of 4% DP in the laboratory. The results showed that the mortality rates of the snails were higher with 4% DP than 4% PG in each dosage (t1 d = 3.60, P < 0.01). The LC50 (s) of 1d, 3 d, 7 d after dusting the molluscicide also showed that the molluscicidal effects of DP were better than PG. The corrected mortality rates were 91.71%, 92.91%, 90.57%, 85.33% and 71.09%, 90.11%, 90.13%, 85.26% on 3 d, 7 d, 15 d, 30 d after dusting with 4% PG and 4% DP, respectively, in the fields. Statistics showed that the mortality rates of snails were higher on 3 d, 7 d after dusting with PG than DP (χ23 d = 731.57, χ27 d = 25.90, P < 0.01), but there were no significant differences between PG and DP on 15d, 30d after dusting (χ215 d = 0.53, χ230 d = 0.01, P > 0.05). CONCLUSIONS: 4% PG has both the adsorption of powder and the penetrability of the granules. The molluscicidal effects of 4% PG and 4% DP are almost the same. However, the drift of the powder was still not effectively controlled. This problem need to be further studied.

5-Aza-CdR can reverse gefitinib resistance caused by DAPK gene promoter methylation in lung adenocarcinoma cells.

To explore the relationship between death associated protein kinase (DAPK) gene promoter methylation and gefitinib resistance in Lung adenocarcinoma cell lines. EGFR-mutation lung adenocarcinoma cell lines PC9 and the gefitinib-resistant with T790M Mutation cell lines PC9/GR were chosen as cell models, and PC9/GR were treated with 5-aza-CdR (1 µmol/L). The experiments were divided into three groups: PC9 group, PC9/GR group and PC9/GR with 5-Aza-CdR pretreatment group. Treat three groups cell with different concentrations gefitinib, the cell proliferation was determined by MTT assay. The apoptotic rates were detected by flow cytometry. The methylation of DAPK gene promoter region was examined by methylation-specific PCR (MSP). The expressions of DAPK protein were detected by Western blot. MTT results showed that the half maximal inhibitory concentration (IC50) of PC9 and PC9/GR cell lines increase from 0.12 µmol/L to 8.52 µmol/L. But after treated with 5-aza-CdR, the IC50 of PC9/GR cell lines decrease to 4.35 µmol/L, and the resistance index (RI) decrease from 71 to 36 (P<0.05). Flow cytometry results showed that the apoptosis rate were 24.80% ± 0.28%, 12.70% ± 0.31%, 19.8% ± 0.15% respectively. MSP results showed that DAPK gene promoter region was un-methylated in PC9 cells and methylated in PC9/GR cells, when treated with 5-aza-CdR, DAPK gene promoter region was partly methylated in PC9/GR cells (P<0.05). Western blot results showed that the levels of DAPK protein were reduced significantly in PC9/GR cell lines compared with PC9, and after treated with 5-aza-CdR, the expression levels of DAPK protein in PC9/GR were increased (P<0.05). In conclusion, DAPK gene promoter methylation may contribute to the downregulation of DAPK gene and protein, and consequently affect the sensitivity of gefitinib in lung adenocarcinoma lines, induced gefitinib resistance. But 5-Aza-CdR can reverse gefitinib resistance by demethylation of DAPK gene promoter.

[Development of national neglect norm for urban primary school students of China].

OBJECTIVE: To develop the national neglect norms for urban primary school students in China. METHODS: According to multi-stage stratified cluster sampling principle, 24 cities of 13 provinces (municipalities) in China were selected during December 1 to 31, 2008. A total of 1491 students in grade 1 - 3 and 2236 students in grade 4 - 6 were selected. Questionnaire was designed by authors and the final norms were determined through several statistical analysis methods, such as item analysis method, factor analysis method, reliability analysis method. The reliability analysis and validity analysis were used to test the stability and reliability of the norms. The evaluation criteria of the scale was determined by the percentile method, then the initial development of the norm was completed. RESULTS: The two questionnaires of grade 1 - 3 and grade 4 - 6 students consisted of 55 and 57 items, respectively, whose item loadings were ranged from 0.301 to 0.687 and 0.321 to 0.730, which met the statistical requirements. For grade 1 - 3 students, the scale's total Cronbach α coefficients was 0.914, the total split-half reliability coefficients was 0.896, the Cronbach α coefficients of four level was above 0.737 except medical and social neglect, split-half reliability was ranged from 0.461 to 0.757; for grade 4-6 students, the scale's total Cronbach α coefficients was 0.916, split-half reliability was 0.883, except social neglect, the Cronbach α coefficients of other level was ranged 0.457 to 0.856, split-half reliability was ranged from 0.500 to 0.798. The total neglect cut-off score of the two scales grade 1-3 and 4-6 were 125 and 155, respectively. CONCLUSION: The structure of two norms was reasonable. The scales have good stability and reliability.

Clinicopathologic significance of ß-catenin and matrix metalloproteinase-2 expression in non-small cell lung cancer.

The purpose of this study was to analyze ß-catenin and matrix metalloproteinase-2 (MMP-2) expression in non-small cell lung cancer (NSCLC) and to investigate the association between their expression and clinicopathologic characteristics of NSCLC patients. Immunohistochemistry was performed to examine ß-catenin and MMP-2 protein expression in 39 resected NSCLC samples and 8 adjacent normal lung tissues. Statistical analysis with SPSS13.0 software was performed to investigate the association between ß-catenin and MMP-2 expression and clinicopathologic features of the patients. Expression of cytosolic ß-catenin in NSCLC tissue was significantly higher than that in normal tissues (P < 0.001). In addition, cytosolic protein expression of ß-catenin in lung squamous cell carcinoma was significantly elevated compared to that in lung adenocarcinoma (P = 0.02). However, cell membrane protein expression of ß-catenin in squamous cell carcinoma was lower than that in adenocarcinoma (P = 0.041). Cytosolic MMP-2 protein expression in NSCLC samples was significantly higher than that in normal tissues (P = 0.002). MMP-2 expression in N (1-2) NSCLC patients was significantly increased relative to N (0) patients (P = 0.019). However, statistical analysis showed no correlation between ß-catenin and MMP-2 expression in NSCLC samples. Collectively, our results show that cytosolic protein expression of ß-catenin in NSCLC samples is increased relative to normal lung tissues. Also, expression of ß-catenin is significantly elevated in squamous cell carcinoma compared to that in lung adenocarcinoma subtypes. Additionally, MMP-2 expression in N (1-2) NSCLC tissues is higher than that in N (0) lung tissue. There is no correlation between ß-catenin and MMP-2 expression in NSCLC, and our study suggests that evaluation of ß-catenin and MMP-2 expression may have potential in diagnosis and progression in patients with NSCLC.

[Output characteristics of non-point phosphorus from a typical small watershed in Yimeng mountainous area under the special rainfall].

The relationships between pollutant concentrations and precipitation-runoff combined with the first and maximum intensity natural rainfall events were monitored at the outlets of the Menglianggu watershed in 2010. The results showed that dissolved organic phosphorus (DOP), dissolved inorganic phosphorus (DIP), total dissolved phosphorus (DP), total particle phosphorus (PP) and total phosphorus (TP) concentrations in the first rainfall were higher than those in the maximum intensity rainfall which was the largest average rainfall intensity, but the output of phosphorusin the first rainfall was all higher than that in the maximum intensity rainfall except dissolved organic phosphorus. The DOP, DIP, DP, PP and TP in two special rainfall events contributed 48%, 81%, 70%, 87% and 81% of the total output of rainy season. DP and TP were mainly composed by DIP and PP respectively. The two special rainfalls have the first flush effect and the maximum intensity rainfall was stronger than the first rainfall, so the analysis of M (V) curve showed that the priority should be given to the interception of corresponding phosphorus under special events. The regression analysis results showed that a linear relation existed between runoff and nutrient output, but there were some differences which the slope of the linear equation of the first rainfall was higher than that of the maximum intensity rainfall. Reducing agricultural practices such as tillage and fertilization during special rainfall period and building canals, ponds are effective ways for mitigating agricultural non-point source pollution.

[A comparative study of survival time of surgery combined with chemotherapy and non-surgical chemotherapy in SCLC].

OBJECTIVE: To compare the survival time of surgery combine with chemotherapy with non-surgery chemotherapy in SCLC. METHODS: 37 SCLC post operation patients were received 4-6 cycles of chemotherapy of "etoposide + cisplatin or carboplatin". 37 SCLC patients were selected be control group, whose age, stage and site of tumor were similar to surgery combine with chemotherapy group, and received 6 cycles of chemotherapy of "etoposide + cisplatin or carboplatin". Two groups were followed up 5 year and compared with the survival time. RESULTS: The 1, 3, 5 year survival rate of surgery combined with chemotherapy group were 72.97%, 35.13%, 21.62%. The 1, 3, 5 year survival rate of control group were 54.05%, 13.51%, 5.41%. The survival rate of stage I (P = 0.01) was significantly different, but stage II (P = 0.06) and stage III (P = 0.836) were no difference between the two group of SCLC. CONCLUSION: It was found that surgery combined with chemotherapy got satisfactory effects in stage I SCLC.

[Treatment of femoral fractures by bone grafting from trochanter and interlocking intramedullary nail fixation].

OBJECTIVE: To discuss the curative effect of bone graft from trochanter with interlocking intramedullary nail for comminute fracture of femur. METHODS: From January of 2009 to January of 2010, fifty patients with comminute fracture of femur were treated by the therapy of bone graft from trochanter with interlocking intramedullary nail, included 35 males and 15 females with an average age of 38.4 years old ranging from 28 to 50 years. According to AO/ASIF classification system, there were 15 cases of type B1, 20 of type B2, 8 of type B3, 7 of type C1. RESULTS: All the patients were followed up for 12 to 24 months, averaged 14.4 months. All the broken femurs knitted in 4 to 6 months, the average period of knit was 5.6 months. The injured legs with normal movement in knees,which had no infection, vascellum-nerve injury and anchylosis in knees. According to Merchan knee rating scale, the results were excellent in 25 cases, good in 20, fair in 3 and poor in 2, and the excellent and good rate was 90%. CONCLUSION: Bone graft from trochanter with interlocking intramedullary nail, which has a virtue of small injury, less blood loss, high rate of bone knit, perfect functional recovery, is a preferable therapy of fresh fracture of femur.

[Anterior cruciate ligament reconstruction with hamstring tendons preserved tibial insertion and an implant fixation technique of hamstring tendon knot and bone bolt press-fit].

OBJECTIVE: To evaluate the feasibility of hamstring autograft anterior cruciate ligament reconstruction with hamstring tendons insertion on tibia preserved and an implant fixation technique of hamstring tendons knot and bone bolt press-fit. METHODS: Twenty cases of injured anterior cruciate ligament were reconstructed. There were 15 males and 5 females with the average age of 22 years old. Ten left knees and 10 right knees were involved. Hamstring tendons were taken, and pretension was performed. Tibial tunnel and femoral tunnel were prepared,and the femoral tunnel was a frame of narrow inside and wide outside. The hamstring tendons knot and bone bolt were pulled inside of femoral tunnel. The tendons distal of tendons knot were brought to pass the femoral tunnel, joint capsule and another tibial tunnel. Then, the tendons distal of tendons knot were tightened together with the part of hamstring tendons of which the insertion were on tibia. After the operation, the knee was fixed at a flexion of 45 degrees by brace. RESULTS: The patients were followed up for 8 to 24 months. The function of troubled knees was evaluated by Lysholm knee functional scale. The average knee score were 61.5 +/- 4.6 and 92.5 +/- 3.7 respectively before and after operation, and the difference was statistically significant (P < 0.05). CONCLUSION: Hamstring tendons insertion on tibia preserved and an implant fixation technique of hamstring tendons knot and bone bolt press-fit was biological fixation for anterior cruciate ligament reconstruction with hamstring. The advantage of this method was avoidance of using high cost material for fixation,which lessened spending for the patients. And it was also benefit for tendon-bone healing.

[Effects of acupuncture on bone metabolism and serum estradiol level in ovariectomy-induced osteoporosis rats].

OBJECTIVE: To observe the effect of acupuncture (Acu) on bone metabolism and serum estradiol (E2) in ovariectomized (OVX) rats for studying its underlying mechanism in treating osteoporosis. METHODS: Forty female SD rats of six months were randomized into sham operation (sham), model, Acu and Diethylstibestrol (DES) groups, with 10 cases in each. Postmenopausal osteoporosis model was established by removing ovaries under anesthesia. In Acu group, bilateral "Dazhu" (BL 11), "Shenshu" (BL 23) and "Pishu" (BL 20) were punctured and stimulated for 30 minutes, once daily for 60 days. Rats of DES group were drenched with saline+DES (22.5 microg/ml) 1 ml/100 g, once daily for 60 days. At the end of experiments, blood samples were collected from femoral artery for assaying serum alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP) contents by using biochemistry, and serum bone gla protein (BGP) and E2 levels by immunoradioassay. RESULTS: Compared with sham group, uterus wet weight, serum E2 content in model group decreased significantly (P < 0.01) while body weight, serum ALP, BGP and TRAP levels in model group increased significantly (P < 0.01, 0.05). Compared with model group, uterus wet weight and serum Ez content in Acu and DES groups increased significantly (P < 0.01); while body weight, serum ALP, BGP and TRAP levels decreased considerably (P < 0.01). No significant differences were found between Acu and DES groups in serum E2, ALP, BGP and TRAP levels (P > 0.05). CONCLUSION: Acupuncture can suppress abnormal increase of body weight and decrease of serum E2 level, and significantly downregulate serum ALP, BGP and TRAP levels in OVX rats, which may contribute to its effect in relieving osteoporosis.

[Posttraumatic stress disorder (psychiatric injury) after road traffic accidents in forensic medicine: a primary study].

OBJECTIVE: Studying forensic medical problem related with RTA leading PTSD and supplying accumulating evidence for psychiatric compensation in court. METHODS: One hundred and fifty six victims of RTA were recruited who applied to court for a costs order. The victims were examined for psychiatric diagnosis by psychiatrists and for rank of impairment by experts in forensic clinical medicine. The self-report psychopathological status and quality of life were also measured. RESULTS: Eighty one victims of 156 (51.92%) fulfilled the criteria for PTSD (ICD-10). Morbidity difference in male and female were significant; The more serious extent of impairment is, the more PTSD'possibility is; The scores in World Health Organization Quality of Life were lower and in SAS and SDS were higher in PTSD group than in non-PTSD group. Acquirement of awarded costs could obviously prevent PTSD. CONCLUSION: The higher PTSD incidence existed in the RTA victims who applied to court for a costs order, and acquirement of awarded costs could obviously prevent PTSD.