Co-delivery nanoparticles of doxorubicin and chloroquine for improving the anti-cancer effect in vitro.

To increase the efficacy of small molecule chemotherapeutic drug (SMCD) and reduce its toxic and side effects, we selected two model drugs doxorubicin (DOX) and chloroquine (CQ). DOX is a SMCD and CQis a chemosensitizer with autophagy inhibition. Poly(lactic-co-glycolic acid) (PLGA) and alpha-tocopherol polyethylene glycol 1000 succinate were chosen as delivery carriers to design and prepare a novel type of drug co-delivery single-nanoparticles by emulsification-solvent volatilisation, named NPDOX+CQ. The physicochemical properties of NPDOX+CQ were characterised. Then A549 cells and A549/Taxol cells were used for the in vitro anti-cancer effect study. At the same time, cellular uptake, intracellular migration and anti-cancer mechanism of nanoparticles were studied. The NPs showed a uniform spherical shape with good dispersibility, and both drugs had good encapsulation efficiency and loading capacity. In all formulations, NPDOX+CQ showed the highest in vitro cytotoxicity. The results showed that NPs could protect drugs from being recognised and excluded by P-glycoprotein (P-gp). Moreover, the results of the mechanistic study demonstrated that NPs were transported by autophagy process after being taken up by the cells. Therefore, during the migration of NPDOX+CQ, CQ could exert its efficacy and block autophagy so that DOX would not be hit by autophagy. Western Blot results showed that NPDOX+CQ had the best inhibition effect of autophagy. It can be concluded that the system can prevent the drug from being recognised and excluded by P-gp, and CQ blocks the process of autophagy so that the DOX is protected and more distributed to the nucleus of multidrug resistance (MDR) cell. The NPDOX+CQ constructed in this study provides a feasible strategy for reversing MDR in tumour cells.

Nature differences of humic acids fractions induced by extracted sequence as explanatory factors for binding characteristics of heavy metals.

The composition and structure of Humic acid (HA) is so heterogeneous that it brings significant barriers to investigate the interaction between HA and heavy metal ions. The isolation of HA with relatively homogeneity is a key to reveal the binding mechanisms between HA and heavy metals. In this work, ten HA fractions (HAs) were obtained by sequential alkali extraction procedure and nature differences of the extracted HAs were considered as explanatory factors for binding characteristics of Cu2+, Pb2+ and Cd2+. The results indicate that more large molecular weight (MW) HA subunits, less carboxyl and phenolic group contents, weaker aromaticity and polarity were measured with increasing extractions, inducing weaker binding capacity of HAs. Ligand binding and bi-Langmuir models indicated that the sorption capacity and binding affinity of earlier extracted HAs were higher than the latter ones. The peak area changes at 3427, 1599, and 619 cm-1 pre- and post-adsorption in FTIR spectra suggested carboxyl, phenolic and nitrogen-containing groups were involved in the adsorption process. At the same time, the peak area difference between HAs and HAs-metal (ΔS) of phenolic groups were 8.22-20.50, 6.81-21.11 and 10.66-19.80% for Cu2+, Pb2+ and Cd2+, respectively, ΔS of carboxyl groups 6.64-17.03, 8.96-16.82 and 9.45-17.85% for Cu2+, Pb2+ and Cd2+, respectively, ΔS of nitrogen-containing groups 0.33-0.48, 0.20-1.38 and 0.31-0.59% for Cu2+, Pb2+ and Cd2+, respectively. ΔS of phenolic and carboxyl groups were larger than those of nitrogen-containing groups, implying that these two groups were the predominant binding sites suppliers for metal ions, which were also supported by the results of correlation analysis. This work is helpful to insight the environmental impacts of natural organic matter and the fate of heavy metals in natural environment.

Digital Anatomical Measurement for Anterolateral Fixation of Middle and Lower Thoracic Vertebrae.

BACKGROUND The key to its successful application is to determine the best entry point for the vertebral screw(s). This study aimed to provide a reference for clinical anterolateral fixation through digital measurement of computed tomography (CT) data to identify relevant anatomical positions in the middle and lower thoracic vertebrae (T4-T12) of 30 adults. MATERIAL AND METHODS We performed digital measurement of anatomical positions in the middle and lower thoracic vertebrae (T4-T12) of 30 adults. ABBREVIATIONS: Left height of vertebral body, LHV; Right height of vertebral body, RHV; Anterior height of vertebral body, AHV; Middle height of vertebral body, MHV; Posterior height of vertebral body, PHV; Superior sagittal diameter of vertebral body, SSDV; Superior transverse diameter of vertebral body, STDV; inferior sagittal diameter of vertebral body, ISDV; Inferior transverse diameter of vertebral body, ITDV; (1) Left (right) height of vertebral body, [L(R)HV]; Anterior (middle, posterior) height of vertebral body [A(M,P)HV]; Superior (inferior) sagittal diameter of vertebral body, [S(I)SDV]; Superior (inferior) transverse diameter of vertebral body, [S(I)TDV]. RESULTS The transverse diameters of vertebral bodies were always larger than the sagittal diameter for 3~4 mm. The distance between 2 vertebrae (interval of 1 vertebra) range were (52-56) mm for T4-T7 and (44-48) mm for T8-T12, and the surgeons could collate these data to choose a suitable stick length. CONCLUSIONS Bone graft should prune into laterigrade cuboid, it can recover A-P and bilateral physiological functions load, and the height of the vertebral body increased from T4 to T12.

Experimental Application of Bone Marrow Mesenchymal Stem Cells for the Repair of Intervertebral Disc Annulus Fibrosus.

BACKGROUND This study provides experimental results on the applicability of bone marrow mesenchymal stem cells (BMSCs) for the repair of intervertebral disc annulus fibrosus in rabbits. MATERIAL AND METHODS Thirty healthy rabbits were randomized into an observation group (n=15) and a control group (n=15). Both groups underwent degeneration of intervertebral disc annulus fibrosus. The observation group was treated with a solution of BMSCs and dexamethasone sodium phosphate, while the control group was treated with dexamethasone sodium phosphate only. RESULTS The two groups were compared for efficacy and pathological conditions after treatment. Both disc height index and level of type II collagen in nucleus pulposus were significantly higher in the observation group than in the control group at 2, 4, 8, and 12 weeks after degeneration (p<0.05 for all comparisons). The percentages of grade 0 and grade 1 were significantly higher in the observation group than in the control group (p<0.05 for both grade 0 and 1 comparisons), while the percentage of grade 4 and grade 5 were significantly lower in the observation group than in the control group (p<0.05 for both grade 4 and 5 comparisons). CONCLUSIONS BMSCs cultured in vitro can effectively repair intervertebral disc annulus fibrosus, which is of positive significance, and thus is clinically recommended.

[Intraoperative oculomotor nerve monitoring during skull base tumor surgery].

OBJECTIVE: To explore the intraoperative oculomotor nerve monitoring during skull base tumor surgery for injury avoidance. METHODS: The study was conducted from January 2006 to January 2012. Monitoring group:35 skull base tumor operations were monitored by neurophysiology (22 males and 13 females). CONTROL GROUP: 26 skull base tumor operations without intraoperative monitoring were treated with microsurgery alone (16 males and 10 females). The prognoses of different groups were evaluated. RESULTS: Monitoring group:2 cases had injury of oculomotor nerve. One of them suffered mild diplopia and recovered one months later. Another one had blepharoptosis and mydriasis, reaction to light was absent and recovered six months later. CONTROL GROUP: 6 cases had injury of oculomotor nerve. Five of 6 cases recovered six months later. And one showed no recovery after a 1-year follow-up. CONCLUSION: Intraoperative monitoring of oculomotor is essential for the protection of oculomotor nerve during skull base tumor surgery.

[Effect of altitudes on the photosynthate accumulation and distribution pattern of Angelic sinensis].

OBJECTIVE: To study the effect of altitudes on the photosynthate accumulation and distribution pattern of Angelic sinensis in Gansu province and provide theontical for its expanding ecological planting region. METHODS: Used field test to study the photosynthate accumulation and distribution pattern of Angelic sinensis on three different altitudinal gradients from 2 300 m to 2 800 m. RESULTS: Before September 25, total photosynthate accumulation were decreased with the increasing of elevation, which amaunt was 176 g/plant, 166 g/plant and 128 g/plant, respectively. The total photosynthate of low-altitude and middle-altitude were significantly higher than that of high-altitude (P < 0.05). After September 25, middle-altitude was significantly higher than the other two altitudes (P < 0.05), respectively, by 13.9% and 11.1%. The photosynthate accumulation rate existed the altitude effects, there was no significant difference between high-altitude (46.7%) and middle-altitude (43.7%), but they were even significantly higher than that of low-altitude (33.1%). The root distribution proportion (> 30%) existed the difference, that of high-altitude was 10 days earlier than the other two altitudes, and later that of high-altitude (about 54%) was higher than the other two altitudes (49.8% - 50.9%), it laid the foundation for yield formation. Yield of Angelic sinensis was as follows: middle-altitude (28.4 g/ plant), high-altitude (26.6 g/plant) and low-altitude (21.8 g/plant). Yield of Angelic sinensis middle-altitude and high-altitude were higher than that of low-altitude, respectively, by 30.2% and 22.2%, and it had a significant difference (P < 0.05), this result was consistent with the photosynthate accumulation rate. CONCLUSION: Altitudinal gradients affect yield formation of Angelic sinensis by changing the photosynthate distribution pattern and dry matter accumulation rate. So by appropriately increasing altitude, the root distribution proportion and yield are improved, this provides theoretical reference for expanding Angelic sinensis planting ecological region.

Copper-Catalyzed Si-H Bond Insertion Reaction of N-Propargyl Ynamides with Hydrosilanes.

Transition-metal-catalyzed Si-H bond insertion reactions are generally limited to stabilized diazo compounds. An efficient copper-catalyzed Si-H bond insertion reaction of N-propargyl ynamides with hydrosilanes is described, allowing practical and atom-economic construction of valuable organosilanes in generally moderate to excellent yields under mild reaction conditions. Notably, this reaction constitutes a new method of Si-H bond insertion reaction involving vinyl cations as key intermediates.

Copper-catalyzed tandem cis-carbometallation/cyclization of imine-ynamides with arylboronic acids.

An efficient copper-catalyzed tandem regioselective cis-carbometallation/cyclization of imine-ynamides with arylboronic acids has been developed. This method leads to a facile and practical synthesis of valuable 2,3-disubstituted indolines in moderate to excellent yields and features a broad substrate scope and wide functional group tolerance. Other significant features of this protocol include the use of readily available starting materials, high flexibility, simple procedure and mild reaction conditions.

[Survey of feeding patterns in 437 infants from Huining County of Gansu Province].

OBJECTIVE: To investigate the feeding patterns of infants in Huining County, Gansu Province, an economically undeveloped county in China, so as to provide scientific evidence for nutritional intervention. METHODS: By cluster sampling based on the area distribution and the economic state, 437 children from 7 different areas in Huining County, Gansu Province were enrolled. A questionnaire on the feeding patterns within 24 months of age was completed by the caregivers. The addition of nutrient supplements (calcium, zinc, ferri, vitamin AD, etc.) during the last year was investigated using the food frequency questionnaires. RESULTS: The average percentages of breast feeding, mixed feeding and artificial feeding were 45.3%, 40.5% and 14.2% respectively. Delayed introduction of complementary food was the most common. Formula powder was added earliest, at age of 6.4 months. The meats were added latest, at age of 17.2 months. Only 66.8% of infants received an addition of complementary food at age of 4-6 months. Grains, vegetables, fruits and eggs were common complementary additional diets, but fishes and shrimps were seldom provided. The proportion of nutrient supplement use was low. The choice of nutrient supplements in urban districts mainly depended on the physicians' suggestions, but in rural area it depended on the caregivers' experience and media advertisements. CONCLUSIONS: The feeding patterns for the infants are not reasonable in Huining County, Gansu Province. It is important to provide nutritional education and reasonable feeding program for caregivers from economically undeveloped areas.

miR-596 suppresses the expression of Survivin and enhances the sensitivity of osteosarcoma cells to the molecular targeting agent anlotinib.

BACKGROUND: Osteosarcoma (OSA), the most common primary bone malignancy, is characterized by a wide spectrum of complicated pathologies and frequent distal metastasis and causes death in adolescents and young adults worldwide. Antitumor drug treatment strategies include various cytotoxic chemotherapy drugs, while molecular targeted therapy for OSA is currently less used. The present work revealed the role played by the miR-596/Survivin axis in affecting the sensitivity of OSA cells to anlotinib, a novel molecular targeting agent. METHODS: By virtual screening, we found that miR-596 might target Survivin by using an online tool (miRDB). RNA levels of miR-596 and Survivin in clinical specimens were examined with qPCR. The effect of miR-596 on anlotinib's antitumor effect was examined with MTT experiments, the subcutaneous tumor model, or the intramuscular tumor model. RESULTS: Overexpression of miR-596 via lentiviral particles repressed the protein level of Survivin in U2OS cells. Transfection of miR-596 enhanced the antitumor effect of anlotinib on U2OS cells or five cell lines derived from OSA patients. CONCLUSION: miR-596 targets Survivin and enhances the antitumor effect of anlotinib on OSA cells.

Differential proteomic analysis to identify proteins associated with beak deformity in chickens.

The beak is the dominant avian facial feature, and beak deformity occurs in 0.5 to 2.5% of some indigenous chicken breeds, resulting in difficulties when eating, drinking, and performing natural behaviors. Previous studies on beak deformity focused largely on candidate molecules associated with skeletogenic development, providing insight into the molecular and genetic underpinnings of beak deformity. The present study was performed to identify candidate proteins related to this malformation in chickens. Three 12-day-old Beijing-You roosters with deformed beaks (D1, D2, and D3) and 3 with normal beaks (N1, N2, and N3) were used, and total beak proteins were isolated and subjected to standard iTRAQ labeling, strong cation-exchange chromatography, and liquid chromatography-tandem mass spectrometry. Mascot 2.3.02 was used to identify and quantitatively analyze proteins. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were used to identify functions and metabolic pathways of differentially expressed proteins, and key proteins were further validated using western blot. A total of 2,370, 2,401, and 2,378 proteins were reliably quantified in 3 biological replicates, among which, 2,345 were common to all, and 92 were differentially expressed between the 2 groups. These included 37 upregulated and 55 downregulated proteins in deformed beaks. Pentraxin-related protein 3, hemopexin, lipoprotein lipase, retinoid-binding protein 7, and biliverdin reductase A were downregulated in all 3 sets, while parvalbumin, peptidyl-prolyl cis-trans isomerase, and ubiquitin-fold modifier 1 were upregulated. Pathway analysis returned no enriched pathways, and western blot validated the iTRAQ results. Parvalbumin and lipoprotein lipase could be firstly selected as key proteins in view of their known functions in regulating the buffering of intracellular free Ca2+ in both cartilage and bone cells and bone mass, respectively. Their potential roles in beak deformity, however, deserve further studies. In summary, the onset of beak deformity could be very complex, and this study will be helpful for future investigation of mechanistic explanation for beak deformity.

[Effects of Fulvic Acid on Absorption and Form Distribution of Heavy Metals on Sediments].

Based on the extracted fulvic acid (FA) from Lake Wuliangsuhai sediments by sequential alkali extraction, this work studied the effects of FA on the adsorption and fraction distribution of heavy metals (HM) on sediments using original sediments and sediments treated with 30% H2O2 as adsorbents. The results showed both organic matter and FA had effects on the HM adsorption onto sediments; The treatments of FA-free conditions and the sediments treated by H2O2 showed relatively strong influence on Cu²âº adsorption, which decreased the Cu²âº adsorption by 17.85%. With the increasing FA addition, the adsorption percentage of HM on both types of sediments showed gradually decreasing trends, with the order of Cu²âº >> Cd²âº > Zn²âº > Pb²âº; when the FA content was more than 5% , FA became the governing factor on the decreasing adsorption percentage of HM. With increasing FA addition, forms distribution of HM showed significant changes in both types of sediments; i. e. FA additions showed significant negative and positive correlations with percentages of metals bound to carbonates and organic matter, respectively, since the FA addition increased the H⁺ concentration of the system, in which H⁺ could activate the metals bound to carbonate from the sediments. As an organophilic weak element, the fraction percentage of Cd bound to organic matter was the lowest with the minimal changes.

[Effects of stubble-standing mode on the grain yield and water use efficiency of wheat and maize in wheat/maize intercropping system].

Wheat/maize intercropping is the main intercropping pattern in the irrigation region of Hexi Oasis, Northwest China, but the traditional intercropping needs much water, making the regional water resource lacked increasingly. In 2010, a field experiment was conducted in the irrigation region of Shiyang River basin oasis, Gansu Province of Northwest China, aimed to study the effects of traditional stubble-burning, stubble-returning, and stubble-standing on the grain yield, water use efficiency (WUE), and economical benefits of wheat and maize in wheat/maize inter-cropping system. Compared with stubble-burning and stubble-returning, stubble-standing increased the grain yield of mono- and intercropped wheat by 7.2% and 5.1% , and 6.2%, 5.1%, and that of mono- and intercropped maize by 4.7% and 2.5%, and 7.2% and 3.3%, and increased the WUE of mono- and intercropped wheat by 20.4% and 16.2%, and 17.9% and 14.6%, and that of mono- and intercropping maize by 16.7% and 10.9%, and 11. 8% and 17.0%, respectively. As for the mono- and intercropped wheat and maize, their average net economical benefits under stubble-burning, stubble- returning, and stubble-standing were 10946, 11471, and 13454 RMB.hm-2, respectively. In considering the grain yield, WUE, and economic benefits, stubble- standing would be the optimal mode of wheat/maize intercropping in the oasis of Hexi irrigation region, Northwest China.

[Effects of different water harvesting modes on alfalfa planting in semi-arid areas of Northwest China].

A field experiment with complete random design was conducted to investigate the effects of different mulching materials [common plastic film (CMR), biodegradable mulch film (BMR), and soil crust (SR)] and different ratios of furrow to ridge (60 cm:30 cm, 60 cm:45 cm, and 60 cm:60 cm) on the runoff efficiency, soil water storage, soil water content, and hay yield and water use efficiency of alfalfa in semiarid areas of the Loess Plateau. The runoff efficiency in treatments SR, BMR, and CMR was 32.0%, 90.7%, and 96.4%, respectively. In the early growth period of alfalfa (from April to June) , the soil water storage between the treatments had no significant difference, but in the late growth period (from July to September), the soil water storage in CMR and BMR was significantly higher than that in SR. The soil water storage in SR was significantly higher than that in traditional planting (TP). At budding stage, the soil water storage in TP, SR, BMR, and CMR was 223.27, 248.56, and 277. 81, and 284.16 mm, respectively. In the whole growth period, the hay yield of alfalfa in TP, SR, BMR, and CMR was 4112.1, 3397.5, 4317.8, and 4523.8 kg x hm(-2), and the water use efficiency was 11.08, 10.48, 14.56, and 14.95 kg x mm(-1) x hm(-2), respectively. The ratio of furrow to ridge had no significant effects on the water use efficiency in the same treatments. When the ratio of furrow to ridge was 60 cm:44 cm, the hay yield in CMR and BMR reached the maximum.

[Window partial laryngectomy for the treatment of stage T2-3 glottic laryngeal carcinoma].

OBJECTIVE: To investigate the operative effect and value of window partial laryngectomy for the treatment of stage T2-3 glottic laryngeal carcinoma. METHODS: From Oct. 2000 to Dec. 2006, the stage T2-3 glottic laryngeal carcinoma of 48 patients by properly selected were performed with window partial laryngectomy and laryngeal reconstruction. Twenty nine males and 19 females were included. Their ages ranged from 43 to 78 years (median 57.0 years). Before operation and in the 6 months after operation, these targets including auditory mental evaluation of hoarse degree, active degree and symmetry of vocal cord, glottic width in the time of quiet breathing, glottic closing degree in the time of phonating, respiratory function and swallowing function were surveyed. In addition, the things of recurrence and metastasis of tumors together with survival time of patients were following investigated. RESULTS: All patients were decannulated successfully and incisions were healed smoothly. No operative complication occurred. Except active degree of vocal cord (P = 0.343), there were respectively significant difference between two group targets of auditory mental evaluation of hoarse degree (all P <0.01), symmetry of vocal cord (P = 0.000), glottic width in the time of quiet breathing (P = 0. 001), glottic closing degree in the time of phonating (P = 0.001) and respiratory function (P=0.001) those were investigated before operation and after operation. The swallowing function wasn't influenced (P= 0.310). There were laryngostenosis in 1 case, recurrence in 1 case (2.1%), cervical lymph node metastasis in 1 case and hepatic metastasis in 1 case. Two cases died. 3-year and 5-year overall survival rate were respectively 96.9% and 88.9%. CONCLUSIONS: The study showed that window partial laryngectomy was successful for treating stage T2-3 glottic laryngeal carcinoma by properly selected. This operation was effective for reducing surgical invasion and beneficial to resume respiratory and vocal function.

[Diagnosis and treatment of serious pediatric airway obstruction].

OBJECTIVE: To investigate the diagnosis and treatment of serious pediatric airway obstruction. METHODS: Seventy three pediatric cases with serious airway obstruction, admitted to Shantou hospital from 1995 to 2005, were analyzed retrospectively. RESULTS: Among the 73 cases, 28 cases (38.4%) with inflammatory disease and 33 cases (45.2%) with foreign body. In addition, 8 cases of laryngeal papilloma, 3 cases of laryngotracheobronchial spasm and 1 case of pharyngeal dysembryoma were also included in this study. The patients who had endotracheal intubation or tracheotomy were 39 cases (53.4%) and 27 cases (36.9%) respectively. Ten cases received non-surgery treatment. Seventy two cases diagnosis was confirmed and the related serious airway obstruction condition got stable within 12 hours. While serious complications occurred in 23 cases (31.5%) and operative complications occurred in 3 cases (4.1%). Four cases ( 5.5%) died and 68 cases (93.2%) were cured. One case refused to be treated. CONCLUSIONS: The condition of serious pediatric airway obstruction patients is usually critical, the related causes could be complicated, quick diagnosis and timely removal of the obstruction factors are imperative.