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Abstract Although sex differences have been reported in patients with clear cell renal cell carcinoma (ccRCC), biological sex has not received clinical attention and genetic differences between sexes are poorly understood. This study aims to identify sex-specific gene mutations and explore their clinical significance in ccRCC. We used data from The Cancer Genome Atlas-Kidney Renal Clear Cell Carcinoma (TCGA-KIRC), The Renal Cell Cancer-European Union (RECA-EU) and Korean-KIRC. A total of 68 sex-related genes were selected from TCGA-KIRC through machine learning, and 23 sex-specific genes were identified through verification using the three databases. Survival differences according to sex were identified in nine genes (ACSS3, ALG13, ASXL3, BAP1, JADE3, KDM5C, KDM6A, NCOR1P1, and ZNF449). Female-specific survival differences were found in BAP1 in overall survival (OS) (TCGA-KIRC, p = 0.004; RECA-EU, p = 0.002; and Korean-KIRC, p = 0.003) and disease-free survival (DFS) (TCGA-KIRC, p = 0.001 and Korean-KIRC, p = 0.000004), and NCOR1P1 in DFS (TCGA-KIRC, p = 0.046 and RECA-EU, p = 0.00003). Male-specific survival differences were found in ASXL3 (OS, p = 0.017 in TCGA-KIRC; and OS, p = 0.005 in RECA-EU) and KDM5C (OS, p = 0.009 in RECA-EU; and DFS, p = 0.016 in Korean-KIRC). These results suggest that biological sex may be an important predictor and sex-specific tailored treatment may improve patient care in ccRCC.

Abstract Utilizing a microfluidic chip with serpentine channels, we inoculated the chip with an agar plug with Neurospora crassa mycelium and successfully captured individual hyphae in channels. For the first time, we report the presence of an autonomous clock in hyphae. Fluorescence of a mCherry reporter gene driven by a clock-controlled gene-2 promoter (ccg-2p) was measured simultaneously along hyphae every half an hour for at least 6 days. We entrained single hyphae to light over a wide range of day lengths, including 6,12, 24, and 36 h days. Hyphae tracked in individual serpentine channels were highly synchronized (K = 0.60-0.78). Furthermore, hyphae also displayed temperature compensation properties, where the oscillation period was stable over a physiological range of temperatures from 24 °C to 30 °C (Q10 = 1.00-1.10). A Clock Tube Model developed could mimic hyphal growth observed in the serpentine chip and provides a mechanism for the stable banding patterns seen in race tubes at the macroscopic scale and synchronization through molecules riding the growth wave in the device.

Abstract We determined the macroscopic limit for phase synchronization of cellular clocks in an artificial tissue created by a “big chamber” microfluidic device to be about 150,000 cells or less. The dimensions of the microfluidic chamber allowed us to calculate an upper limit on the radius of a hypothesized quorum sensing signal molecule of 13.05 nm using a diffusion approximation for signal travel within the device. The use of a second microwell microfluidic device allowed the refinement of the macroscopic limit to a cell density of 2166 cells per fixed area of the device for phase synchronization. The measurement of averages over single cell trajectories in the microwell device supported a deterministic quorum sensing model identified by ensemble methods for clock phase synchronization. A strong inference framework was used to test the communication mechanism in phase synchronization of quorum sensing versus cell-to-cell contact, suggesting support for quorum sensing. Further evidence came from showing phase synchronization was density-dependent.

Introduction Patient education is a crucial component in the management of chronic kidney disease (CKD); however, many education programmes do not tailor to patients’ disease-related belief and emotional reactions. According to Leventhal’s self-regulation model, such beliefs and reactions, known as illness perceptions, are a part of patients’ understanding of their disease and treatment-related actions. The existing research of illness perception in the field of nephrology has been heterogenous in terms of study outcomes and in study population; and the evidence of how illness perceptions affect behavioural and decisional outcomes is limited. This scoping review aims to map and summarise the evidence of published literature on illness perception among patients with CKD not yet treated by dialysis and kidney failure.Methods and analysis This study bases its approach on Joanna Brigg’s Institute Guidelines on scoping review methods. The search strategy was developed together with a medical information specialist. Searches will be performed on acceptance of publication in the following databases: PubMed, EMBASE, PsycINFO via Ovid, Scopus and the Cumulative Index to Nursing and Allied Health Literature. Searches will be run without incorporating a date restriction in order to capture content from the databases’ inception to present day. Search terms including ‘illness perception’, ‘kidney disease’ and ‘kidney failure’ will be screened in titles and abstracts. Two independent researchers will screen the abstracts and full text for full eligibility. We will include studies focusing on illness perception of patients with CKD with estimated glomerular filtration rate

Using a microfluidics device, fluorescence of a recorder (mCherry or mVenus) gene driven by a clock-controlled gene-2 promoter (ccg-2p) was measured simultaneously on over 1000 single cells of Neurospora crassa every half hour for 10 days under each of varied light and temperature conditions. Single cells were able to entrain to light over a wide range of day lengths, including 6, 12, or 36 h days. In addition, the period of oscillations in fluorescence remained remarkably stable over a physiological range of temperatures from 20 °C to 30 °C (Q10 = 1.00-1.07). These results provide evidence of an autonomous clock in most single cells of N. crassa. While most cells had clocks, there was substantial variation between clocks as measured by their phase, raising the question of how such cellular clocks in single cells phase-synchronize to achieve circadian behavior in eukaryotic systems at the macroscopic level of 107 cells, where most measurements on the clock are performed. Single cells were placed out of phase by allowing one population to receive 6 or 12 h more light before lights out (D/D). The average phase difference was reduced in the mixed population relative to two unmixed control populations.

Background: Protease-activated receptors (PARs) can be stimulated by thrombin and other proteases generated by periodontal pathogens. Activation of PARs in gingival fibroblasts (GFs) can modulate wound healing and inflammatory responses in gingival tissues.Methods: The mRNA expression of PARs and early responsive genes in GFs and other oral cells was studied by reverse transcription-polymerase chain reaction. Western blotting was used to study the activation of p38 and cAMP responsive element binding protein (CREB)/activating transcription factor (ATF)-1 as well as Ras.Results: GFs, dental pulp cells, and buccal mucosal fibroblasts expressed PAR-1 and -3 receptors, whereas gingival keratinocytes expressed PAR-1 and -2 receptors. Stimulation of GFs by thrombin rapidly activated Ras signaling and the phosphorylation of CREB/ATF-1 and p38. Thrombin also stimulated the expression of c-fos in GFs within 1 hour of exposure. Stimulation of c-jun mRNA expression showed biphasic responses with two peaks after 1 and 8 hours of exposure. Elevated c-myc expression in GFs by thrombin was noted after 2 hours of exposure. Moreover, the stimulation of c-fos and c-myc mRNA expression by thrombin can be attenuated by D-Phe-Pro-ArgCH(2)Cl, a serine-proteinase inhibitor.Conclusions: PAR activation during gingival wounding or inflammation may stimulate Ras-CREB/ATF-1 signaling and c-fos, c-jun, and c-myc expression. This might be due to the proteinase activity of thrombin. These signaling events are important for wound healing and inflammatory responses in gingival tissues. [ABSTRACT FROM AUTHOR]

The high prevalence of acquiring skin wounds, along with the emergence of antibiotic-resistant strains that lead to infections, impose a threat to the physical, mental, and socioeconomic health of society. Among the wide array of wound dressings developed, hydrogels are regarded as a biomimetic soft matter of choice owing to their ability to provide a moist environment ideal for healing. Herein, neutral glycol chitosan (GC) was cross-linked via imine bonds with varying concentrations of dibenzaldehyde-terminated polyethylene glycol (DP) to give glycol chitosan/dibenzaldehyde-terminated polyethylene glycol hydrogels (GC/DP). These dynamic Schiff base linkages (absorption peak at 1638 cm -1 ) within the hydrogel structure endowed their ability to recover from damage as characterized by high-low strain exposure in continuous step strain rheology. Along with their good injectability and biodegradability, the hydrogels exhibited remarkable inhibition against E. coli , P. aeruginosa , and S. aureus . GC/DP hydrogels demonstrated high LC50 values in vivo using zebrafish embryos as a model system due to their relative biocompatibility and a remarkable 93.4 ± 0.88% wound contraction at 30-dpw against 49.1 ± 3.40% of the control. To the best of our knowledge, this is the first study that developed injectable glycol chitosan/dibenzaldehyde-terminated polyethylene glycol self-healing hydrogels for application in wound healing with intrinsic bacteriostatic properties against the three bacteria.

Inosine 5'-monophosphate dehydrogenase (IMPDH; EC1.1.1.205) is the rate-limiting enzyme of GMP biosynthesis. The inhibition of IMPDH limits the growth and survival of tumors. Based on the systematic summary of clinical IMPDH inhibitors, 38 acrylamide derivatives with differently substituted indoles at the 3-position as the core scaffold were designed and synthesized. In addition, the actions of these compounds at the enzyme and cellular levels were evaluated. An MTT assay with different kinds of cells was used to assess the cytotoxic activities of compounds 14e and 14n, which displayed potent hIMPDH2 inhibitory activities (IC 50  = 4.207 and 2.948 μM, respectively). Biological evaluation indicated that target compounds 14e and 14n displayed the most significant effects on SW480 human colon cancer cells (IC 50  = 15.34 ± 0.06 and 15.31 ± 0.09 μM, respectively), and it was determined that these compounds are effective and valuable IMPDH inhibitors for cancer intervention., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Described herein is an efficient strategy for assembling a new library of functionalized polycyclic purinium salts with a wide range of anions through Rh III -catalyzed C-H activation/annulation of 6-arylpurine nucleosides with alkynes under mild reaction conditions. The resulting products displayed tunable photoluminescence covering most of the visible spectrum. Mechanistic insights delineated the rhodium catalyst's mode of action. A purinoisoquinolinium-coordinated rhodium(I) sandwich complex was well characterized and identified as the key intermediate.

In this paper, carbon aerogel (CA)-polyaniline (PANI) composites were prepared and first applied in the study of H 2 S gas sensing. Here, 1 and 3 wt% of as-obtained CA powder were blended with PANI to produce composites, which are denoted by PANI-CA-1 and PANI-CA-3, respectively. For the H 2 S gas-sensing studies, the interdigitated electrode (IDE) was spin-coated by performing PANI and PANI-CA composite dispersion. The H 2 S gas-sensing properties were studied in terms of the sensor's sensitivity, selectivity and repeatability. IDE coated with PANI-CA composites, as compared with pristine PANI, achieved higher sensor sensitivity, higher selectivity and good repeatability. Moreover, composites that contain higher loading of CA (e.g., 3 wt%) perform better than composites with lower loading of CA. At 1 ppm, PANI-CA-3 displayed increased sensitivity of 452% at relative humidity of 60% with a fast average response time of 1 s compared to PANI.

This experiment treated perilla seeds with different concentrations of NaCl solution to enrich and purify their rosmarinic acid (RosA). The results showed that low concentrations of salt (0-20 mmol/L) promoted seed germination, while high concentrations (> 20 mmol/L) inhibited germination. When the salt concentration was 20 mmol/L, the germination rate was the highest. The content of RosA in germinated perilla seeds was 3.5 mg/g, which was 3.5 times as much as that in the seeds without germination. The RosA was purified using NK-109 macroporous resin and its adsorption kinetics, isotherms and thermodynamics were determined. The adsorption kinetics showed that the adsorption behavior of RosA in NK-109 resin conformed to the pseudo-second-order kinetic model. The model for RosA in the NK-109 resin exhibited Langmuir adsorption based on a spontaneous exothermic process according to its adsorption thermodynamics, which included both physical and chemical adsorption. The optimized process conditions were as follows: the loading concentration of 0.04 mg/mL, loading volume of 40 mL, 70% methanol as the eluent with the volume of 60 mL, and the purity of RosA was 42.1%.

Electrochemically driven, Cp*Ir(III)-catalyzed regioselective annulative couplings of benzoic acids with alkynes have been established herein. The combination of iridium catalyst and electricity not only circumvents the need for stoichiometric amount of chemical oxidant, but also ensures broad reaction compatibility with a wide array of sterically and electronically diverse substrates. This electrochemical approach represents a sustainable strategy as an ideal alternative and supplement to the oxidative annulations methodology to be engaged in the synthesis of isocoumarin derivatives.

Neurilemmoma, also known as schwannoma or neurinoma, is a tumor that originates from neural sheath Schwann cells. Giant neurilemmomas derived from the retroperitoneum have rarely been reported. We herein describe a woman with a giant retroperitoneal neurilemmoma that was initially incorrectly diagnosed as an inflammatory abdominal mass. The tumor extended from the patient's hypogastrium to her pelvic cavity and measured 20 × 15 × 10 cm. The tumor was excised via laparotomy and diagnosed as a retroperitoneal neurilemmoma through histological and immunohistochemical examination. Although rare, particularly in the giant form, neurilemmoma should be considered as an important differential diagnosis in patients with a retroperitoneal tumor or inflammatory abdominal mass. Complete excision should be considered for the potential cure of giant retroperitoneal neurilemmomas.

The controversy of adjuvant radiotherapy of meningiomas is at least partially due to the insufficient understanding on meningioma cells' response to irradiation and the shortage of radiosensitivity-promotion methods. MicroRNA-221 and microRNA-222 were identified as critical regulators of radiosensitivity in several other tumors. However, their effect in meningiomas has yet to be confirmed. Therefore, the malignant meningioma IOMM-Lee cells were adopted, transfected with microRNA-221/222 mimics or inhibitors, and irradiated with different dosages. The effects of radiation and microRNA-221/222 were then assessed in vitro and in vivo . Radiation dose increases and microRNA-221/222 downregulation synergistically inhibited cell proliferation and colony formation, prevented xenograft tumor progression, and promoted apoptosis, but antagonistically regulated cell invasiveness. Pairwise comparisons revealed that only high-dose radiations (6 and 8 Gy) can significantly promote cell invasiveness in comparison with unirradiated counterparts. Further comparisons exhibited that downregulating the microRNA-221/222 expression can reverse this radiation-induced cell invasiveness to a level of untransfected and unirradiated cells only if cells were irradiated with no more than 6 Gy. In addition, this approach can promote IOMM-Lee's radiosensitivity. Meanwhile, we also detected that the dose rate of irradiation affects cell cycle distribution and cell apoptosis of IOMM-Lee. A high dose rate irradiation induces G0/G1 cell cycle arrest and apoptosis-promoting effect. Therefore, for malignant meningiomas, high-dose irradiation can facilitate cell invasiveness significantly. Downregulating the microRNA-221/222 level can reverse the radiation-induced cell invasiveness while enhancing the apoptosis-promoting and proliferation-inhibiting effects of radiation and promoting cell radiosensitivity., (Copyright © 2020 Zhang, Song, Huo, Wang, Zhang, Hao, Jia, Kong, Jia, Wu, Xu, Jia and Zhang.)

Background: Osteosarcoma is a primary malignant bone tumor that frequently occurs in adolescents and children, its high aggressiveness and rapid metastasis often resulting in poor prognoses. In previous studies, Prazosin has been shown to possess anti-proliferative properties against prostate cancer and glioblastoma cells. In our study, we investigated Prazosin's underlying mechanisms and its effects on the biological behaviors of osteosarcoma cells., Methods: Osteosarcoma cell lines MG63 and 143B were treated with different concentrations of Prazosin, and a CCK8 assay assessed its effect on cell viability. Colony formation, Transwell and flow cytometry assays were used to examine its effects on cell proliferation, cell migration, and cell invasion and apoptosis, respectively. The expression of relevant proteins was then examined using western blotting., Results: Our data showed that Prazosin dose-dependently reduced the viability of MG63 and 143B cells and significantly inhibited their clonogenic ability. Moreover, Prazosin attenuated the cell migration and invasion abilities of MG63 and 143B cells when compared with the NC group. It also accelerated cell apoptosis in mitochondrial pathways by regulating Bcl-2/Bax axis and caspase 3. Furthermore, Prazosin treatment inactivated the Akt/mTOR pathway by down-regulating Akt and mTOR phosphorylation (p-Akt, p-mTOR) and the expression of P70 and cyclin D1., Conclusions: Our data highlights the fact that Prazosin inhibits cell growth, inhibits the motility of osteosarcoma cells, and promotes apoptosis, suggesting that Prazosin is a potential anti-cancer agent in osteosarcoma therapy., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at http://dx.doi.org/10.21037/tcr.2019.09.03). The authors have no conflicts of interest to declare., (2019 Translational Cancer Research. All rights reserved.)

Objective: This study was conducted to analyze the clinical characteristics and pituitary function of patients with primary empty sella (PES). Methods: The clinical data from 123 hospitalized adult patients with PES from January 2010 to May 2016 were retrospectively studied. Results: (1) The average age of the 123 (male 43, female 80) PES patients was (59.2±13.6) years (ranging 24-92 years), among whom 61% patients were in the age group between 50-69 years. (2) The symptoms of the patients included fatigue (56.1%), headache (34.1%), nausea and vomiting (17.9%), gonadal dysfunction (17.1%), visual disturbance (5.7%) and hypopituitarism crisis (3.3%). (3) Hypopituitarism was found in 66 of the 123 patients. Among them, 36.6%, 31.7% and 17.1% were central hypoadrenalism, hypogonadism, and hypothyroidism, respectively.The percentage of hypopituitarism in complete PES was significantly higher than that in partial PES ( P <0.05). (4) Sixteen patients were concomitant with other autoimmune diseases including 11 patients with Graves' disease and 2 with Cushing's syndrome due to adrenal adenoma. Conclusions: The incidence of hypopituitarism in PES was 53.7%, in which the pituitary-adrenal axis hypofunction was more common. An overall evaluation of the pituitary function was essential for the patients who had headache and fatigue, or with suspected PES. The patients with hypopituitarism should be given hormone replacement therapy in time and followed up afterword.

On the basis of the molecular understanding that DMSO has a trigonal pyramidal geometry with one highly polarized sulfinyl group and two hydrophobic methyl groups, we used NMR technology to rationalize why DMSO can act as antifreeze for aqueous solutions. A series of DMSO-water, DMSO-methanol, and acetone-water binary solutions with various molar ratios were tested in specified low temperature conditions. The freezing test results indicated that only DMSO-water solutions with the DMSO-water molar ratio (n DMSO ) in a specific range can form apparent ultralow freezing temperature solutions. Among all DMSO-water solutions, the apparent freezing temperature lower than -130 °C was obtained for n DMSO values of 0.25, 0.30, and 0.35, respectively. Multinuclear NMR chemical shifts, 1 H diffusion experiment results, and viscosity measurements suggested that molecular rearrangement resulted in the formation of the water-core DMSO-shell aggregation unit in DMSO-water solutions. The weak methyl-methyl London dispersion forces among water-core DMSO-shell aggregation units can explain the apparent ultralow freezing temperatures of these DMSO-water solutions.

Alzheimer's disease (AD) is the most common neurodegenerative disorder of the brain. It causes the slow progressive loss of cognitive functions that ultimately leads to dementia and death in the elderly. The etiology and mechanism of late-onset AD (LOAD) are poorly understood, and genetic factors might play an important role in the development of AD. The aim of this study was to investigate the association between common polymorphisms in TGF-β1 with LOAD in a Chinese Han population. Two single nucleotide polymorphisms in TGF-β1 (rs1800469 and rs1982073) were genotyped in 202 patients with sporadic LOAD and 225 control subjects using polymerase chain reaction restriction fragment length polymorphism. Our results showed that rs1800469 in TGF-β1 were significantly associated with LOAD. The frequencies of the AC genotypes of rs1800469 were significantly higher in the LOAD patients than in the control subjects (42.5% vs 28.6%; P = 0.001). The minor allele (C) frequency was significantly higher in patients with LOAD than in control subjects (30.7% vs. 21.0%; P = 0.001). The genotypes and allele of rs1982073 in TGF-β1 were also significantly associated with LOAD. The frequency of the TG genotype of rs1982073 was significantly higher in the LOAD patients than in the control subjects (38.1% vs. 27.1%; P = 0.013). The minor allele (G) frequency was significantly higher in patients with LOAD than in control subjects (22.2% vs. 16.7%; P = 0.032). These results suggest that common variants in TGF-β1 might contribute to the development of LOAD in the Chinese population.

The discrimination of tea varieties plays very important role in the production and sale of tea. It is of great significance for the study of a fast, easy and simple method for the identification of tea varieties. The combination of infrared spectroscopy detection technology and fuzzy clustering algorithm is one of the most effective and practical techniques in the detection of tea varieties. To realize the rapid discrimination of tea varieties, a novel generalized noise clustering (NGNC) was proposed based on fast generalized noise clustering (FGNC). Euclidean distance in the objective function of FGNC was replaced with the pth power of Euclidean distance and clustering accuracy was being improved. Emeishan Maofeng; high quality Leshan trimeresurus and low quality Leshan trimeresurus were prepared as the research object and the infrared reflectance (IR) spectra of tea samples were collected with FTIR-7600 infrared spectrometer. Firstly, the high-dimensional IR spectra of tea samples were reduced by principal component analysis (PCA). Secondly, linear discriminant analysis (LDA) was used to extract the discriminant information from the low-dimensional data. Finally, FGNC and NGNC were performed to identify tea varieties. The experimental results showed that in comparision with FGNC, NGNC has higher clustering accuracy, better cluster centers and faster convergence speed. Infrared spectroscopy coupled with NGNC, PCA and LDA could cluster IR spectra of tea samples quickly and correctly, which provided a new method and new idea for identifying tea varieties based on infrared spectroscopy and fuzzy clustering.

Based on anisometric noble-metal nanocrystals, a universal fabrication protocol for preparing 3D supercrystals with controlled orientation on a chip has been developed. A comparison of the surface-enhanced Raman scattering (SERS) behavior of 3D nanorod supercrystals aligned vertically and parallel to the chip indicates that the SERS-enhancing ability and reproducibility of the former is superior to the latter. The 3D nanorod supercrystals aligned vertically to the chip have been utilized as highly sensitive SERS substrates for the label-free discrimination of Gram-positive and -negative bacteria. Furthermore, to strengthen the stability of the supercrystal substrate for assays of bacteria in biosamples, a coating of the antibiotic vancomycin can dramatically increase adhesion of bacteria on a nanointerface and simultaneously improve the SERS response of bacteria to achieve a layer-by-layer assembled, stable, and reliable biosensor for bacteria., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Although cryptococcosis mainly occurs in the central nervous system and lungs in immunocompromised hosts, it can involve any body site or structure. Here we report the first case of primary cryptococcosis of a lumbar vertebra without involvement of the central nervous system or lungs in a relatively immunocompromised individual with rheumatoid arthritis and scleroderma., Case Presentation: A 40-year-old Chinese woman with rheumatoid arthritis diagnosed 1 year beforehand and with a subsequent diagnosis of scleroderma was found to have an isolated cryptococcal infection of the fourth lumbar vertebra. Her main complaints were severe low back and left leg pain. Cryptococcosis was diagnosed by CT-guided needle biopsy and microbiological confirmation; however, serum cryptococcal antigen titer was negative. After 3 months of antifungal therapy with fluconazole the patient developed symptoms and signs of scleroderma, which was confirmed on laboratory tests. After taking fluconazole for 6 months, the progressive destruction of the lumbar vertebral body had halted and the size of an adjacent paravertebral mass had decreased substantially. On discharge symptoms had resolved and at an annual follow-up there was no evidence of recurrence on the basis of symptoms, signs or imaging investigations., Conclusion: Although cryptococcosis of the lumbar vertebra is extremely rare, it should be considered in the differential diagnosis for patients with lumbar vertebral masses to avoid missed diagnosis, misdiagnosis and diagnostic delay. Early treatment with antifungals proved to be a satisfactory alternative to surgery in this relatively immunocompromised patient. Any residual spinal instability can be treated later, once the infection has resolved.

Objective: We explored the clinical values of (11)C-choline ((11)C-CHO) PET in optimization of target volume delineation and treatment regimens in postoperative radiotherapy for brain gliomas., Methods: Sixteen patients with the pathological confirmation of the diagnosis of gliomas prior to receiving radiotherapy (postoperative) were included, and on whom both MRI and CHO PET scans were performed at the same position for comparison of residual tumors with the two techniques. (11)C-CHO was used as the tracer in the PET scan. A plain T1-weighted, T2-weighted and contrast-enhanced T1-weighted imaging scans were performed in the MRI scan sequence. The gliomas' residual tumor volume was defined as the area with CHO-PET high-affinity uptake and metabolism (V(CHO)) and one with MRI T1-weighted imaging high signal intensity (V(Gd)), and was determined by a group of experienced professionals and clinicians., Results: (1) In CHO-PET images, the tumor target volume, i.e., the highly metabolic area with a high concentration of isotopes (SUV 1.016-4.21) and the corresponding contralateral normal brain tissues (SUV0.1-0.62), was well contrasted, and the boundary between lesions and surrounding normal brain tissues was better defined compared with MRI and (18)F-FDG PET images. (2) For patients with brain gliomas of WHO Grade II, the SUV was 1.016-2.5; for those with WHO Grades III and IV, SUVs were >26-4.2. (3) Both CHO PET and MRI were positive for 10 patients and negative for 2 patients. The residual tumor consistency between these two studies was 75%. Four of the 10 CHO-PET-positive patients were negative on MRI scans. The maximum distance between V(Gd) and V(CHO) margins was 1.8 cm. (4) The gross tumor volumes (GTVs) and the ensuing treatment regimens were changed for 31.3% (5/16) of patients based on the CHO-PET high-affinity uptake and metabolism, in which the change rate was 80% (4/5), 14.3 % (1/7) and 0% (0/4) for patients with WHO Grade II III, and IV gliomas, respectively., Conclusion: Our data demonstrate that difference exists between CHO PET and MRI by which to judge and identify residual tumor for patients with brain gliomas. CHO PET is considered to be a supplementary diagnostic approach for MRI. Biological tumor target volume (BTV) displayed in the CHO PET images is useful in determining or delineating the radiotherapy target volume and making decisions in selecting treatment regimens. Tumor target volume may be defined more accurately and rationally when the CHO PET is combined with MRI., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Traditional strategies for establishing shRNA expression constructs are inefficient, error-prone, or costly. We describe a simple approach that overcomes these drawbacks. Briefly, the sense and antisense strands of the short hairpin RNA coding sequence are segmented into two parts, respectively, at asymmetric sites. The four resulting short oligonucleotides are synthesized. Each oligonucleotide is annealed with its opposite, resulting in a double-stranded fragment with sticky termini at both ends. The two fragments so generated can be easily spliced by simple ligation to reconstitute the full-length short hairpin RNA coding sequence which can then be cloned into an appropriately restricted vector.

Objective: To investigate the relationship between gastrointestinal dyskinesis and histologic changes of gastrointestinal myenteric plexus cholinergic and nitrergic neurons in STZ-induced diabetic rats., Methods: 45 SD rats were randomly divided into control group, diabetic group and insulin group. 16 weeks after diabetic model established, gastrointestinal motility of rats was measured and histologic changes of myenteric plexus cholinergic neuron and nitrergic neuron was observed., Results: Compared with control group, gastrointestinal motility of diabetic group was markedly slow (P < 0.01), the myenteric plexus cholinergic neuron counting of gastric antrum and small intestine were significantly decreased (6.6 +/- 2.9 vs 15.7 +/- 3.8 15.6 +/- 10.3 vs 22.6 +/- 7.4, P < 0.01), yet the number of nitrergic neuron only markedly reduced in gastric antrum (5.3 +/- 1.2 vs 11.8 +/- 2.2, P < 0.01). The gastrointestinal mobility, gastric antrum nitrergic neuron and small intestine cholinergic neuron counting of insulin group were markedly higher than that of diabetic group (P < 0.05), yet lower than that of control group (P < 0.01)., Conclusion: The gastrointestinal dyskinesis of STZ-induced diabetic rats might be associated with lesions of gastrointestinal myenteric plexus cholinergic neuron and nitrergic neuron. Insulin intensive therapy can partly ameliorate diabetic gastrointesternal dyskinesis.

Aim: To evaluate the antioxidant and pro-oxidant properties of chlorhexidine (CHX)., Methodology: The scavenging and generation of reactive oxygen species (ROS) by CHX in the presence or absence of saturated Ca(OH)(2) solutions was evaluated. The reaction emitted chemiluminescence in the presence of lucigenin thus was determined by a luminometer to evaluate the levels of ROS production. Changes in DNA conformation were analysed by agarose gel electrophoresis. Paired Student's t-test was used to compare the difference between groups., Results: Chlorhexidine (0.00002-0.02%) effectively scavenged 56-88% of the superoxide radicals generated by the xanthine/xanthine oxidase reaction. Through analysis of PUC18 DNA conformation changes, CHX was shown to be a mild scavenger of hydroxyl radicals generated by H(2)O(2) plus FeCl(2). However, CHX (>0.083%) decreased the mobility of PUC18 plasmid DNA with potential production of DNA-DNA cross-link and severe DNA breaks (presence of DNA smear) at further higher concentrations. Furthermore, CHX induced ROS production including H(2)O(2) and superoxide radicals in 0.1N NaOH (pH = 12.76) or Ca(OH)(2) (pH = 12.5) solutions., Conclusion: Chlorhexidine exhibited both antioxidant and pro-oxidant properties under different conditions. These events are possibly involved in the killing of root canal and periodontal microorganisms when CHX and Ca(OH)(2) were used in combination or separately. Potential genotoxicity and tissue damage when extruded into the periradicular tissue and at higher concentrations should be considered during periodontal and endodontic practice.

The amplification and cloning of promoters are regularly employed procedures to study the mechanism of gene regulation. In the present study we developed a method to construct T-vectors used for the direct and unidirectional cloning and analysis of promoters. These so-called T-vectors pEGFP-T and pGL3-T were derived from their parent promoterless vectors pEGFP-1 and pGL3-basic, respectively. To construct the T-vectors, an AhdI recognition site within the Ampr gene in pGL3-basic was silent mutated using overlap extension PCR. Then, a specially designed AhdI cassette was cloned into the respective parent vectors. The procedures of the T-vector construction involved a strategy to minimize the background of nonrecombinant transformants and to eliminate reverse orientation of the PCR products into the T-vectors. The cloning efficiencies of the two T-vectors were both above 85% when tested with a PCR product amplified from a sequence that was pre-confirmed to be able to initiate transcription, and moreover, the constructs harbored the inserts in a desired orientation at a >90% rate. In transient transfection assays, we demonstrated these T-vectors are functional. Thus, the present study provides an easy method to construct a series of T-vectors used for promoter characterization.

Objective: To study the therapeutic effects of Rhizoma Drynariae and estrogen on osteoporosis in ovariectomized (OVX) rats., Methods: Fifty-five female rats were randomly divided into 5 groups, the normal control (normal),the sham operated (sham), the model, the estrogen, and the Rhizoma drynariae (RD) groups; ovariectomized rats were used as postmenopausal osteoporosis model. The changes of morphology and dynamic parameters in different groups were determined by bone histomorphometry., Results: Compared with the model group, the trabecular volume (TBV/TTV) , trabecular thickness (MTPT) and density (MTPD) in the other four groups were significantly increased, while the trabecular template spacing (MTPS) and the ratio of trabecular surface to trabecular volume (TBS/TBV) significantly decreased (P <0. 05); and the osteoid surface (TOS), single label surface [Sfract (s) ] ,double label surface [Sfract (d) ] and bone formation rate (Svf) also decreased,while osteoid maturation period (OMP) increased in the latter four groups. No significant difference of cortical width (MCW) was found between these 5 groups. Compared with the normal and sham groups, TOS, Sfract ( s) , Sfract ( d) , Svf in the estrogen and RD groups increased significantly, while OMP decreased; no significant difference was found in other parameters., Conclusion: Rhizoma Drynariae has the similar effect with estrogen in maintaining normal trabecular structure and connection by inhibiting the increased bone turnover of postmenopausal osteoporosis.

Objective: To assess the possible antioxidant and anti-inflammatory activity of cyanidin from cherries on adjuvant induced arthritis (AA) in SD rats., Method: Arthritis was induced by the complete Freud's adjuvant in male Sprague Dauley rats and assessed based on paw swelling. Rats were randomly divided into normal group (NM), adjuvant arthritis group (AA) and three cyanidin-treated groups in high dosage (HA), middle dosage (MA), and low dosage (LA). The morphological changes in the hind limbs were conducted under a light microscope. We detected glutathione (GSH) in whole blood and malonaldehyde (MDA), superoxide dismutase (SOD), total antioxidative capacity (T-AOC) activity in serum by special kits to assess the antioxidant effects of cyanidin on AA. Moreover, the prostaglandin E2 (PGE2) levels in paw tissues were determined by radioimmunoassay and TNF-alpha levels in serum were determined using ELISA kits specific for rat., Result: The cyanidin could protect against the paws swelling in AA rats. From the day 14 after AA induction, the swellings of the cyanidin treated groups at high dosage and low dosage were significantly reduced compared with the model group (P < 0.05, 0.01). Histological examination of sections through the hind limbs revealed alleviation of inflammatory reaction in the joint after the treatment. The cyanidin at high and low dosage could increase the GSH, SOD activity and T-AOC levels in whole blood or serums and decrease MDA in AA rats (P < 0.01). The cyanidin could decrease the PGE2 levels in paw tissues and the TNF-alpha levels in serum at high and low dosages (P < 0.01)., Conclusion: The cyanidin could protect against the paws swelling in AA rats, and alleviate the inflammatory reaction in the joint, and the mechanism might be via the increase activity of GSH, SOD and T-AOC that improve the total antioxidative capacity and scavenge the free radicals, perhaps as a result of that the levels of the PGE2 in paw tissues and TNF-alpha contents in serum were decreased. The results suggest that the cyanidin from cherries could be one of the potential candidates for the alleviation of arthritis.

Aim: To explore the effect of Gui Zhi decoction on enteric mucosal immune in type II collagen-induced arthritis (CIA) in DBA mice., Methods: Eighty DBA/1, weighing 18-22 g, were randomly divided into four groups with 20 in each group: control group, CIA group, treatment groups at high dosage and low dosage (GZH and GZL). CIA was induced by immunization with type II collagen (CII) emulsified with equal complete adjuvant at 0.1 mg CII each mouse. Blood lymphocyte suspension was screened for CD4 and CD8 expression using a flow cytometry, the CD4 and CD8 and secretory IgA (sIgA)-positive cells in enteric lamina propria tested with immunohistochemical staining. Tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1)-beta, and IL-6 concentrations in serum were assayed with RIA., Results: Gui Zhi decoction can lower the arthritic scores and decrease the occurrence of arthritis. The CD4, CD8, and sIgA-positive cells in CIA mice are less than in control mice, and in Gui Zhi decoction at high dosage could restore the lowered CD4- and CD8-positive cells in lamina propria, and at both high and low dosages could increase the lowered sIgA-positive cells in lamina propria, even still lower than in normal mice. In periphery, the CD4 cells in periphery are higher in CIA mice than in control mice, and Gui Zhi decoction at high and low dosages could decrease the CD4 and CD8 cells. Also, Gui Zhi decoction at high dosage could decrease the IL-6 and TNF-alpha concentration in serum., Conclusion: Gui Zhi decoction can lower the arthritic scores and decrease the incidence of CIA in mice, and the mechanism is in part regulating enteric mucosal immune.

Objective: To observe the effects of Tripterygium polyglycosides (TWP) on mucous immune function in rat models of arthritis induced respectively with collagen-II induced arthritis (CIA) & adjuvant arthritis (AA)., Methods: CIA and AA model rats were induced by immunization with collagen II emulsified with complete Freund's adjuvant and complete Freund's adjuvant respectively and treated with TWP. Rats' mucus, systemic immunological indexes (peripheral subsets of T cells), local inflammatory factors (IL-6, TNF-alpha, COX-2,and NF-kappaB, etc. ) were observed., Results: In CIA model group, CD4+ in Peyer's Patch (PP), peripheral CD4+ and CD8+ positive T cells all raised, while in the AA model group, CD4+ lowered and CD8+ raised on PP, with both subsets increased. Effects of TWP on T lymphocyte subsets in PP and blood of the two models were different. High leveled IL-6, TNF-alpha, COX-2 and NF-kappaB expression could be seen in both model groups, and these inflammatory media could be inhibited by TWP., Conclusion: There exist similarities and differences between the two models in aspects of mucus immune response and effect of TWP on them.

Aim: To evaluate the correlation between CD4, CD8 cell infiltration in gastric mucosa, Helicobacter pylori (H pylori) infection and symptoms or the assemblage of symptoms in cases with chronic gastritis., Methods: Biopsy samples at the gastric antrum were obtained from 62 patients with chronic gastritis. CD4 and CD8 cell infiltration was evaluated by immunohistochemical assays on frozen sections of the biopsy samples. Fifteen symptoms referring to digestion-related activity and non-digestion related activity were observed. The correlation between lymphocyte infiltration and each symptom or symptom assemblage was analyzed by logistic regression and K-mean cluster methods., Results: CD4 cell infiltrations in gastric mucosa were much more in patients with H. pylori infection, while CD8 cell infiltrations were similar in patients with or without H. pylori infection. Logistic regression analysis showed that the symptoms including heavy feeling in head or body (t = 2.563), and thirst (t = 2.478) were significantly related with CD4 cell infiltration in gastric mucosa (P<0.05), and cool limbs with aversion to cold were related with CD8 cell infiltration (t = 2.872, P<0.05). Further analysis showed that non-digestive related symptom assemblage could increase the predicted percentage of CD4 and CD8 cell infiltration in gastric mucosa, including lower CD4 infiltration by 12.5%, higher CD8 infiltration by 33.3%, and also non-H. pylori infection by 23.6%. K-means cluster analysis of all symptoms and CD4 and CD8 cell infiltration in gastric mucosa showed a similar tendency to increase the predicted percentage of CD4, CD8 cell infiltration and H. pylori infection., Conclusion: Based on correlation between the gastric mucosa lymphocyte infiltration, H. pylori infection and clinical symptoms, symptoms or symptomatic assemblages play an important role in making further classification of chronic gastritis, which might help find a more specific therapy for chronic gastritis.

Traditional Chinese medicine, including herbal medicine and acupuncture, as one of the most important parts in complementary and alternative medicine (CAM), plays the key role in the formation of integrative medicine. Why do not the modern drugs targeting the specificity of diseases produce theoretical effects in clinical observation? Why does not the traditional Chinese medicine targeting the Zheng (syndrome) produce theoretical effects in clinic? There should have some reasons to combine Western medicine with Chinese herbal medicine so as to form the integrative medicine. During the integration, how to clarify the impact of CAM theory on Western medicine has become an emergent topic. This paper focuses on the exploration of the impact of theory of traditional Chinese medicine on the therapy of diseases in Western medicine.

Objective: To clarify the effects of nylestriol on microarchitecture and interleukin-6 (IL-6) mRNA expression in tibial bone in ovariectomized rats., Methods: 30 female rats were randomly allocated into 3 groups: sham, OVX and nylestriol-treated group. Nylestriol-treated group were ovariectomized, then fed with nylestriol for 3 months and the bone mineral density (BMD) was measured in lumbar vertebra by dual energy x-ray absorptiometry. After sacrifice of the animal, bone histomorphometric parameters were measured to study the changes in bone microarchitecture, and RT-PCR was performed to detect the expression of IL-6 mRNA in bone tissue., Results: BMD was significantly reduced, while IL-6 mRNA level elevated in the OVX group compared with the sham group. Static histomorphometric data showed that the trabecular bone volume, mean trabecular plate thickness and density were reduced while the mean trabecular plate space elevated remarkably in the OVX group in comparison with that in the sham group. As for dynamic parameters, trabecular osteoid surface, tetracyclin labeled surface and bone turnover rate were increased while osteoid maturation rate decreased significantly in the OVX group compared with the sham group. BMD, IL-6 mRNA expression and bone histomorphometric parameters were improved in nylestriol-treated rats., Conclusion: Nylestriol plays an important role in maintaining bone volume and improving bone microarchitecture by markedly inhibiting bone turnover and bone resorption, which might be to some degree attributed to reduced IL-6 expression.

The main nutritional limitation of maize used for feed is the content of protein that is digestible, bioavailable and contains an amino acid balance that matches the requirements of animals. In contrast, milk protein has good digestibility, bioavailability and amino acid balance. As an initial effort to create maize optimized as a source of swine nutrition, a codon-adjusted version of a gene encoding the milk protein porcine alpha-lactalbumin was synthesized. Maize expression vectors containing this gene under the control of the Ubi-1 promoter and nos 3' terminator were constructed. These vectors were used to transform maize callus lines that were regenerated into fertile plants. The alpha-lactalbumin transgenes were transmitted through meiosis to the sexual progeny of the regenerated plants. Porcine alpha-lactalbumin was detected in callus and kernels from transgenic maize lines that were transformed by two constructs containing the 27-kDa maize gamma-zein signal sequence at the 5' end of the synthetic porcine alpha-lactalbumin coding sequence. One of these constructs contained an ER retention signal and the other did not. Expression was not observed in kernels or callus from transgenic maize lines that were transformed by a construct that does not contain an exogenous protein-targeting signal. This suggests that the signal peptide might play an important role in porcine alpha-lactalbumin accumulation in transgenic maize kernels.

Background & Objective: Multi-drug resistance(MDR) is the most important failing reason of tumor chemotherapy, the substance that P-glycoprotein(P-gp), glutathione S-transferase-pi(GST-pi), Topoisomerase II (Topo II) etc was the fundamental substances producing MDR. Gastric carcinoma is the relatively familiar malignancy, the report was relatively fewness that the expression of P-gp, GST-pi and Topo II by means of immunohistochemistry were associated with in the gastric carcinomas of previously untreated patients. This study was designed to investigate the expression and significance of P-gp, glutathione S-transferase-pi(GST-pi) and Topoisomerase II(Topo II) in gastric carcinomas., Methods: Seventy-five human gastric carcinomas of previously untreated patients were studied for the expression of P-gp, GST-pi and Topo II by means of immunohistochemistry., Results: The expression of P-gp, GST-pi and Topo II were 76.0%, 85.3% and 68.0% in gastric carcinoma tissues. There was no significant relationship between the expression levels and age, sex, location of tumor and depth of tumor invasion. Positive rate was higher in glandular-form carcinoma than that in mucoid carcinoma, signet-ring cell carcinoma and poorly differentiated carcinoma (P < 0.05). There was a significant relationship between the lymph node involvement and the higher expression of P-gp, GST-pi, lower expression of Topo II., Conclusion: There is a correlation between the expression of P-gp, GST-pi, Topo II and histologic subtypes, the expression may also related to the differentiation and metastasis of the gastric carcinomas.

To study the effect of hypoxia on humoral immunity function of rat and Ochotona curzoniae (pika), the specific antibody production to novel antigen IgG and immunoresponse to sheep red blood cell (hemolysin forming) were measured. The results show that hypoxia at altitude of 5 km and 7 km for 10 d resulted respectively in 10.3% (P < 0.05) and 21.9% (P < 0.05) decrement in hemolysin formation in rats, as compared with the control group kept at 2.3 km. When the rats were secondarily immunized and kept at the same hypoxia for 10 d, the reduction in hemolysin formation was 4.2% (P < 0.05) and 4.6% (P < 0.05) for the two respective altitudes. These changes, however, were not found in pikas. When rats were immunized two days before hypoxia, 5 km hypoxia for 5 d and 8 d failed to suppress hemolysin formation. Intracerebroventricular (i.c.v.) injection of CRF (1.0 microgram/rat), decreased hemolysin formation and production of IgG by 8.6% (P < 0.05) and 14.0% (P < 0.05) respectively, but intraperitoneal (i.p.) injection of CRF (1.0 microgram/rat) had no effect. On the other hand, i.c.v. injection of CRF receptor antagonist (alpha-helical CRF (9-41), 50 micrograms/rat) prior to 7 km hypoxia caused a hypoxia-induced suppression of IgG production from 24.2% to 12.1% (P < 0.05). Adrenalectomy in rats lowered hemolysin formation by 6.6% (P < 0.05). The above results demonstrate that hypoxia suppresses humoral immunity function and alters initial antigen processing probably through an increase of CRF in the central nervous system.