Synthesis, Antifungal Activity, 3D-QSAR, and Molecular Docking Study of Novel Menthol-Derived 1,2,4-Triazole-thioether Compounds.

A series of novel menthol derivatives containing 1,2,4-triazole-thioether moiety were designed, synthesized, characterized structurally, and evaluated biologically to explore more potent natural product-based antifungal agents. The bioassay results revealed that at 50 µg/mL, some of the target compounds exhibited good inhibitory activity against the tested fungi, especially against Physalospora piricola. Compounds 5b (R = o-CH3 Ph), 5i (R = o-Cl Ph), 5v (R = m,p-OCH3 Ph) and 5x (R = α-furyl) had inhibition rates of 93.3%, 79.4%, and 79.4%, respectively, against P. piricola, much better than that of the positive control chlorothalonil. Compounds 5v (R = m,p-OCH3 Ph) and 5g (R = o-Cl Ph) held inhibition rates of 82.4% and 86.5% against Cercospora arachidicola and Gibberella zeae, respectively, much better than that of the commercial fungicide chlorothalonil. Compound 5b (R = o-CH3 Ph) displayed antifungal activity of 90.5% and 83.8%, respectively, against Colleterichum orbicalare and Fusarium oxysporum f. sp. cucumerinum. Compounds 5m (R = o-I Ph) had inhibition rates of 88.6%, 80.0%, and 88.0%, respectively, against F. oxysporum f. sp. cucumerinu, Bipolaris maydis and C. orbiculare. Furthermore, compound 5b (R = o-CH3 Ph) showed the best and broad-spectrum antifungal activity against all the tested fungi. To design more effective antifungal compounds against P. piricola, 3D-QSAR analysis was performed using the CoMFA method, and a reasonable 3D-QSAR model (r2 = 0.991, q2 = 0.514) was established. The simulative binding pattern of the target compounds with cytochrome P450 14α-sterol demethylase (CYP51) was investigated by molecular docking.

[Analysis of spectral characteristics of oil film on water based on wavelet transform].

The diagnostic features are the basis to detect and characterize the oil film on water through optical remote sensing. This work shows the results of lab spectral measurements of light diesel oil with thickness ranged 1.0 - 127 microm. A wavelet transform were performed to the reflectance, and the singularity (388-393 nm) was explored as the indicators of oil film thickness. The results indicate that the reflectance of light diesel oil film is higher than that of water in the range from 350-2 500 nm. There is a reflectance peak near 388 nm when the thickness of oil film is larger than 6 microm, however, no distinguished features could be recognized when oil films were thinner than 6 microm. The wavelet coefficients of the fifth decomposition level by applying Daubechies 4 (db4) mother wavelets proved successful for identifying the singularity of oil film's reflectance spectra and its accurate position. With the thickness lager than 6 microm, the detail coefficients performed an abrupt change within the range of 388-393 nm, and became more violent while oil films' thickness increased. This research demonstrated that oil films on water with different thickness could be distinguished based on wavelet detail coefficients, with important implications for detection of oils on water using UV and short wave optical remote sensing.

Bioinformatics analysis of rabbit haemorrhagic disease virus genome.

BACKGROUND: Rabbit haemorrhagic disease virus (RHDV), as the pathogeny of Rabbit haemorrhagic disease, can cause a highly infectious and often fatal disease only affecting wild and domestic rabbits. Recent researches revealed that it, as one number of the Caliciviridae, has some specialties in its genome, its reproduction and so on. RESULTS: In this report, we firstly analyzed its genome and two open reading frameworks (ORFs) from this aspect of codon usage bias. Our researches indicated that mutation pressure rather than natural is the most important determinant in RHDV with high codon bias, and the codon usage bias is nearly contrary between ORF1 and ORF2, which is maybe one of factors regulating the expression of VP60 (encoding by ORF1) and VP10 (encoding by ORF2). Furthermore, negative selective constraints on the RHDV whole genome implied that VP10 played an important role in RHDV lifecycle. CONCLUSIONS: We conjectured that VP10 might be beneficial for the replication, release or both of virus by inducing infected cell apoptosis initiate by RHDV. According to the results of the principal component analysis for ORF2 of RSCU, we firstly separated 30 RHDV into two genotypes, and the ENC values indicated ORF1 and ORF2 were independent among the evolution of RHDV.

Rescue of Newcastle disease virus from cloned cDNA using an RNA polymerase II promoter.

A new system was developed to improve the efficiency and simplify the procedure of recovery of Newcastle disease virus (NDV) from cloned cDNA. A full-length cDNA clone of mesogenic NDV vaccine strain Mukteswar was assembled from five subgenomic cDNA fragments and cloned into a plasmid allowing transcription driven by cellular RNA polymerase II. The full-length viral cDNA was flanked by hammerhead ribozyme (HamRz) and hepatitis delta virus ribozyme (HdvRz) sequences, resulted in the synthesis of antigenomic RNA with exact termini. Without supplying T7 RNA polymerase, infectious NDV could be generated efficiently in some eukaryotic cell lines by simultaneous transcription of antigenomic RNA from the full-length plasmid and expression of NP, P and L proteins from helper plasmids introduced by cotransfection. The efficiency of recovery with the conventional T7 promoter system based on BRS-T7 cells and the cytomegalovirus (CMV) promoter system was compared, and the results demonstrate that the new system facilitates the generation of recombinant NDV and more efficient than the T7 rescue system using BRS-T7.

Suppression of KIF22 Inhibits Cell Proliferation and Xenograft Tumor Growth in Colon Cancer.

Background: Kinesin family member 22 (KIF22) is known as a regulator of cell mitosis and cellular vesicle transport. The alterations of KIF22 are associated with a series of tumors; however, its possible role in the progression of colon cancer is still unclear. Materials and Methods: This retrospective research collected 82 paired tissues with colon cancer. KIF22 protein and mRNA expression levels were detected by immunohistochemistry assays and Immunoblot assays, respectively. Short hairpin RNA (shRNA) plasmids were used to suppress the expression of KIF22 in HCT116 and HT29 cells, and the silencing efficiencies of shRNA plasmids targeted KIF22 were detected by quantitative PCR assays and immunoblot assays. In addition, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assays and xenograft tumor growth assays were performed to observe cell proliferation in vitro and in vivo. Results: In human colon cancer tissues, the expression level of KIF22 was increased and correlated with clinical pathological features, including tumor stage and clinical stage (p = 0.034, and p = 0.015, respectively). Suppression of KIF22 inhibited cell proliferation and xenograft tumor growth. Conclusion: KIF22 might play an important role in the regulation of cell proliferation in colon cancer and might therefore serve as a promising therapeutic target.

Reverse transcription-loop-mediated isothermal amplification assay for rapid detection of hepatitis E virus.

The one-step single-tube betaine-free reverse transcription (RT)-loop-mediated isothermal amplification assay was developed for rapid diagnosis of hepatitis E virus. This assay amplified the target gene in less than 45 min (even as short as 20 min) under isothermal conditions at 63 degrees C, and the sensitivity of this assay was 100-fold greater than that of RT-PCR. This assay demonstrated a detection limit of 0.045 fg (nine copies/reaction).

High expression of RAB38 promotes malignant progression of pancreatic cancer.

Ras­Related Protein Rab­38 (RAB38), which belongs to the RAB family, is involved in the biogenesis of lysosome­related organelles and defense against certain microbial infections. However, the clinical significance and potential function of RAB38 in pancreatic adenocarcinoma remain unclear. In the present study, an immunohistochemical assay was performed to analyze the expression of RAB38 in pancreatic adenocarcinoma tumor specimens from 82 patients, and the clinicopathological characteristics and survival rate of these patients were further examined. To validate the role of RAB38 in tumors, the effect of RAB38 on tumor cell proliferation, migration and invasion was assessed by establishing RAB38 knockdown cell lines. Reverse transcription­quantitative polymerase chain reaction and western blotting were used to examine the expression levels of proteins associated with the cancer cell behavior. In addition, the inhibitory effect of RAB38 silencing on pancreatic cancer was examined in mice. The immunohistochemistry results revealed that RAB38 was upregulated and positively correlated with the grade of progression in pancreatic adenocarcinoma patients. Further investigation indicated that RAB38 downregulation significantly suppressed the proliferation, migration and invasive capacity of pancreatic cancer cells, as well as decreased the expression levels of Ki67, proliferating cell nuclear antigen, and matrix metalloproteinases 2 and 9. RAB38 silencing also inhibited the development of pancreatic cancer in vivo. Taken together, a high level of RAB38 was significantly associated with the malignant phenotypes of pancreatic cancer, suggesting that RAB38 may serve as a novel biomarker and a potential therapeutic target for pancreatic cancer.

Who should not go high: chronic disease and work at altitude during construction of the Qinghai-Tibet railroad.

From 2001 to 2005, a new railroad linking Beijing with Lhasa was built by more than 100,000 workers, of whom 80% traveled from their lowland habitat to altitudes up to 5000 m to work on the railroad. We report on the medical conditions of 14,050 of these altitude workers, specifically with regard to preexisting illness. All subjects were seen at low and high altitude. Average age was 29.5 +/- 7.4 (SD) yr, range 20 to 62 yr; 98.8% of the subjects were men and 1.2% were women. Overall incidence of AMS upon first-time exposure was 51%, that of HACE 0.28%, and that of HAPE 0.49%. About 1% of the subjects were hypertensive before altitude exposure. Those with blood pressure >or=160/95 were excluded from employment at altitude. Altitude exposure led to a greater increase of blood pressure in hypertensives compared to normotensives. On prealtitude screening prevalence of cardiac arrhythmias was 0.33%. Since the majority of these were rather benign and occurring in young and otherwise healthy subjects, we allowed altitude employment. Follow-up at altitude was uneventful. Subjects with coronary heart disease and diabetes were excluded from altitude employment. Obesity was a risk factor for acute mountain sickness and for reduced work performance at altitude. Overweight subjects lost more weight during their altitude stay than subjects with normal weight. Altitude exposure was a risk factor for upper gastrointestinal tract bleeding, especially in combination with alcohol, aspirin, and dexamethasone intake. Asthmatic subjects generally did better at altitude compared to low altitude, with the exception of one subject who experienced an asthma episode from pollen exposure. In conclusion, careful evaluation of preexisting chronic illness and risk factors allowed prevention of altitude deterioration of a preexisting health condition, all the while allowing subjects with some specific conditions to work and live at altitude without problems.

Complete genomic sequence of transmissible gastroenteritis virus TS and 3' end sequence characterization following cell culture.

The complete genome sequence of transmissible Gastroenteritis virus (TGEV) strain TS, previously isolated from Gansu province, was cloned and compared with published sequence data from other TGEV strains. Phylogenetic tree analysis based on the amino acid and nucleotide sequences of the S gene showed that the TGEV strains were divided into 3 clusters. TGEV TS showed a close evolutionary relationship to the American Miller cluster but had a 5' non-translated region (NTR) sequence closely related to the American Purdue cluster. Continued culture in different cell types indicated that TGEV TS virulence could be attenuated after fifty passages in Porcine kidney (PK-15) cells, and that the Porcine kidney cell line IB-RS-2 (IBRS) was not suitable for culture of the TGEV strain TS.

Altitude illness in Qinghai­Tibet railroad passengers.

It takes ≈24 h to travel the ≈3000-km-long Qinghai­Tibet railroad of which 85% is situated above 4000 m with a pass at 5072 m. Each year about 2 million passengers are rapidly exposed to high altitude traveling on this train. The aim of this study was to quantify the occurrence of altitude illness on the train. Three subject groups were surveyed: 160 Han lowlanders, 62 Han immigrants living at 2200 to 2500 m, and 25 Tibetans living at 3700 to 4200 m. Passengers reached 4768 m from 2808 m in less than 1.5 h, after which 78% of the passengers reported symptoms, 24% reaching the Lake Louise criterion score for AMS. AMS incidence was 31% in nonacclimatized Han compared to 16% in Han altitude residents and 0% in Tibetans. Women and older subjects had a slightly greater risk for AMS. Most cases of AMS were mild and self-limiting, resolving within days upon arrival in Lhasa. Some cases of more severe AMS necessitated medical attention. To curb the health risk of rapid travel to altitude by train, prospective travelers should be better informed, medical train personnel should be well trained, and staged travel with 1 to 2 days at intermediate altitudes should be suggested to nonacclimatized subjects.

[Effects of intermittent hypoxic exposure on the parameter of erythrocyte and serum hypoxia inducible factor-1 alpha and erythropoietin levels].

AIM: To investigate the effects of intermittent hypoxic exposure and normoxic convalescence on the parameter of erythrocyte and serum hypoxia inducible factor-1 alpha (HIF-1alpha) and erythropoietin (EPO) levels. METHODS: Rat models of intermittent hypoxic exposure were established, combined with the clinical research on volunteers experiencing the intermittent plateau work. Blood samples for red blood cell (RBC) counts, hemoglobin (Hb) and hematocrit (HCT) were collected, serum HIF-1alpha and EPO levels were measured using enzyme linked immunosorbent assay. RESULTS: RBC counts, Hb concentration and HCT were significantly higher than the normoxic group (P < 0.05), after exposure of rats to hypoxia from 7 to 28 days. Compared with the normoxic group, serum HIF-1alpha levels were higher in the group of IH3, 7, 14 days, and EPO had a corresponding increase in the group of IH3, 7 days. Then, a decrease was observed in parameter of erythrocyte and serum HIF-1alpha and EPO levels after 14 days normoxic convalescence treat. In volunteers studies, RBC counts in 8 months group and Hb concentration in 2 years group were significantly higher than the plain group (P < 0.05). Compared with the plain group, EPO had no significant differences in any of plateau group. CONCLUSION: Intermittent hypoxic exposure can enhance serum hypoxia inducible factor-1 alpha and erythropointin levels and the generation of red blood cells, which leads to an increase in hemoglobin concentration and hematocrit. The results have changed with the hypoxic exposure period prolonged. Normoxic convalescence after intermittent hypoxic exposure can make the related indexes reduced, and contribute to the organism recovery.

[The effect of insulin on the spatial learning and memory abilities of the Alzheimer's disease model rats].

AIM: To investigate the mechanism through which insulin affect the learning and memory abilities of the Alzheimer's disease-like rats. METHODS: Okadaic acid (OA) was injected into the CA1 region of the rat hippocampus and the insulin was injected into the lateral cerebral ventricle of the rats. The learning and memory abilities of the rats were assessed through Morriswater maze behavioral test, and the expressions of nicotinic acetylcholine receptors and GFAP were observed by Westem blotting and immunohistochemistry, respectively. RESULTS: Compared with the control rats, the abilities of learning and memory were lowered significantly (P < 0.01) and the expressions of the nicotinic acetylcholine receptors were decreased and the GFAP positive astrocytes were increased greatly in the model rats (P < 0.05). In the rats injected with insulin, it was found that their learning and memory abilities were improved significantly (P < 0.01) and that the expression of the nicotinic acetylcholine receptors were increased and GFAP positive astrocytes were decreased obviously (P < 0.05), as compared with the model rats. CONCLUSION: Insulin is able to enhance the learning and memory abilities of the Alzheimer's disease-like rats, possibly by improving the function of the acetylcholine system and decreasing the astrocytes proliferation in the brain.