Dissection of the antitumor mechanism of tetrandrine based on metabolite profiling and network pharmacology.

RATIONALE: Tetrandrine, the Q-marker in Stephaniae Tetrandrae Radix, was proven to present an obvious antitumor effect. Until now, the metabolism and antitumor mechanism of tetrandrine have not been fully elucidated. METHODS: The metabolites of tetrandrine in rats were profiled using ultra-high-performance liquid chromatography coupled with time-of-flight mass spectrometry. The potential antitumor mechanism of tetrandrine in vivo was predicted using network pharmacology. RESULTS: A total of 30 metabolites were characterized in rats after ingestion of tetrandrine (10 mg/kg), including 0 in plasma, 7 in urine, 11 in feces, 9 in liver, 8 in spleen, 4 in lung, 5 in kidney, 5 in heart, and 4 in brain. This study was the first to show the metabolic processes demethylation, hydroxylation, and carbonylation in tetrandrine. The pharmacology network results showed that tetrandrine and its metabolites could regulate AKT1, TNF, MMP9, MMP2, PAK1, and so on by involving in proteoglycan tumor pathway, PI3K-Akt signaling pathway, tumor pathway, MAPK signaling pathway, and Rap1 signaling pathway. CONCLUSIONS: The metabolism features of tetrandrine and its potential antitumor mechanism were summarized, providing data for further pharmacological validation.

Investigation of the effects of high cervical spinal cord electrical stimulation on improving neurological dysfunction and its potential mechanism in rats with traumatic brain injury.

To explore the effects of high cervical spinal cord electrical stimulation (cSCS) on the recovery of neurological function and its possible mechanism in rats with traumatic brain injury (TBI). 72 rats were randomly divided into: (1) a sham group; (2) a traumatic brain injury (TBI) group; (3) a TBI+cSCS group; (4) a LY294002+TBI+cSCS group. The degree of neurological dysfunction was evaluated by modified Neurological severity score (mNSS). The pathological changes of the brain tissue in the injured area were observed by HE staining, and the apoptosis of neuron cells were observed by TUNEL staining. The expressions of BDNF and VEGFmRNA were detected by polymerase chain reaction (PCR), and the expressions of p-AKT, AKT, Bcl-2, Bax and caspase-3 proteins were detected by western blot. Compared with that of the TBI and LY294002+TBI+cSCS groups, the mNSS of the TBI+cSCS group were significantly lower on day 3 and 7 ( P <0.05). Compared with that in the TBI and LY294002+TBI+cSCS groups, the apoptosis of neuron cells in the TBI+cSCS group decreased significantly ( P < 0.05). Compared with the TBI and LY294002+TBI+cSCS group, the expression of Bcl-2 protein increased and the expressions of Bax and Caspase-3 proteins decreased in the TBI+cSCS group ( P < 0.05). Compared with that in the TBI and LY294002+TBI+cSCS groups, the intensity of p-Akt/Akt in the TBI+cSCS group increased ( P < 0.05). We found that cSCS had a protective effect on neuron cells after craniocerebral injury and could improve neurological dysfunction in rats, the mechanism of which might be that cSCS made the PI3K/Akt pathway more active after TBI.

Morphology and concentration of smoke from fluorinated ethylene propylene wire insulation in microgravity under forced airflow.

The production of smoke particles from the jet bursting flame caused by overheating fluorinated ethylene propylene (FEP) wire insulations was investigated. Experiments examining the morphology and volume fraction of the fractal smoke particle aggregates with forced airflow were conducted in a 3.5s drop tower. Gravity level and forced flow were shown to have significant hydrodynamic effects on the pathlines and fractal aggregation of the smoke particles, thus the residence time-dependent flame shape, particle size and concentration have obvious changes. For cases in still air, compared with normal gravity, the jet flame in microgravity has a spherical shape, the mean primary particle and aggregate gyration radius are bigger due to longer residence time, but the fractal dimension maintains at about 1.79, similar to that in the normal gravity level; the calculated smoke volume fraction is also bigger. For cases with force flow in microgravity, the mean primary particle diameter, the mean aggregate gyration radius, and soot volume fraction all decrease with increasing forced flow due to decreasing residence time.

[Study on chlorinated disinfection byproducts and the relevant health risk in tap water of J City].

J City lies in the downstream of Taihu Lake and its water source was micro-polluted by the well-developed industry and agriculture inside the city and in the upper stream. Tap water of J City is characterized as high concentrations of organics and ammonia nitrogen, and chlorinated disinfection byproducts (CDBPs), which has drawn many public concerns for the health risk. Tap water was sampled in May, August, October of 2012 and January of 2013. Four trihalomethanes (THMs) and five haloacetic acids (HAAs) were determined with the gas chromatography. Results revealed that THMs accounted for 88.1% of the sum of THMs and HAAs, with higher concentrations in May, August and January (39.34, 50.37 and 28.02 microg x L(-1), respectively) while obviously lower in October (19.19 microg x L(-1)), which were significantly higher than that of HAAs (2.58-4.02 microg x L(-1)). After boiled for three minutes, THMs were removed over 92.3% but HAAs were largely increased. The health risk of CDBPs was then calculated based on the health risk assessment model recommended by the USEPA. The health risk caused by carcinogenic CDBPs was within a range of 3.1 x 10(-6) - 7. 3 x 10(-6) in the tap water, all over the recommended level of 1 x 10(-6), but after boiled, the value significantly decreased to 7.9 x 10(-7), which is below the recommended level. The health risk caused by non-carcinogenic CDBPs absolutely increased from 2.1 x 10(-11) to 3.4 x 10(-9) after boiled, which is below the reference value of 10(-5).

[Normal values and influencing factors of fractional exhaled nitric oxide for Chinese healthy adults].

OBJECTIVE: To evaluate the normal values of fractional exhaled nitric oxide (FENO) for Chinese adults. METHODS: A total of 200 non-smoking and healthy Chinese adults were recruited according to the inclusion and exclusion criteria. They received lung function and allergen skin prick tests at the Outpatient Department of China-Japan Friendship Hospital from February to August 2011. There were 100 males and 100 females. FENO test was performed according to the standard procedures of American Thoracic Society/European Respiratory Society (ATS/ERS) recommendation. The FENO levels of healthy non-smokers were tested by the NIOXMINO analyzer. All variables, such as age, gender, height, weight, forced expiratory volume at 1 second (FEV(1))% predicted and FEV(1)/FVC, were included for multiple regression analyses. RESULTS: The FENO level of healthy non-smokers was (14.2 ± 5.6)×10(-9) mol/L with the 95% normal range of (3.2 - 25.2)×10(-9) mol/L. The value of FENO was significantly higher in males than females ((15.6 ± 5.5) vs (12.9 ± 5.4)×10(-9) mol/L, t = 3.48, P = 0.001). The FENO level of the subjects aged 60 - 80 years was (15.7 ± 5.4)×10(-9) mol/L and it was higher than those aged 18 - 60 years ((13.8 ± 5.6)×10(-9) mol/L, t = 2.04, P = 0.040). Multiple regression analysis showed that gender was an independent influencing factor of FENO level. CONCLUSIONS: The FENO level of non-smoking and healthy Chinese adults is similar with that of the westerners. Gender is an independent influencing factor of FENO level.

[Study of plasmid-mediated 16S rRNA methylase genes and drug-resistant transferability of Acinetobacter baumannii isolated from burn ward].

OBJECTIVE: To investigate the drug-resistance of Acinetobacter baumannii (Ab) isolated from patients in burn ward, and study the incidence of 16S rRNA methylase genes mediated high-level aminoglycoside drug-resistance and its mechanism of transfer. METHODS: A total of 40 Ab clinical isolates were collected from burn ward in Gansu Province People's Hospital from May 2006 to Dec. 2007. The sensitivity of Ab for 20 antibiotics were determinated by K-B agar diffusion. The minimal inhibitory concentrations (MIC) of amikacin, gentamicin, tobramycin, netilmicin, isepamicin and kanamycin against Ab strains were determinated by agar dilution. Five kinds of 16S rRNA methylase genes including armA, rmtA, rmtB, rmtC, rmtD were amplified by PCR, the positive PCR-products were purified and sequenced, and the plasmid were extracted by alkaline lysis. The transferability of drug-resistance were determinated by conjugation and plasmid transformation tests. RESULTS: The drug-resistance rates of Ab against six aminoglycosides antibiotics was 72.5%, 72.5%, 70.0%, 67.5%, 70.0%, 70.0%, respectively. Twenty five strains were resistant to six aminoglycosides antibiotics (62.5%), among which 10 isolates were armA-positive (40.0%); rmtA, rmtB, rmtC and rmtD-positive isolates were not found. Ten transformants and 10 conjugates showed high-level resistance against aminoglycosides antibiotics, all of which the value of MIC > or = 256 microg/mL carried armA gene. CONCLUSIONS: The drug-resistance of Ab clinical isolates have high drug-resistance. 16S rRNA methylases gene exists in Ab and locates in plasmid chromosome.

[Gas pipeline leak detection based on tunable diode laser absorption spectroscopy].

The principle of tunable diode laser absorption spectroscopy and harmonic detection technique was introduced. An experimental device was developed by point sampling through small multi-reflection gas cell. A specific line near 1 653. 7 nm was targeted for methane measurement using a distributed feedback diode laser as tunable light source. The linearity between the intensity of second harmonic signal and the concentration of methane was determined. The background content of methane in air was measured. The results show that gas sensors using tunable diode lasers provide a high sensitivity and high selectivity method for city gas pipeline leak detection.

Recombinant interferon-alpha2b poly(lactic-co-glycolic acid) microspheres: pharmacokinetics-pharmacodynamics study in rhesus monkeys following intramuscular administration.

AIM: Investigation into pharmacokinetic-pharmacodynamic properties of interferon- alpha (IFN-alpha)2b-loaded poly(lactic-co-glycolic acid) (PLGA) microspheres (MS) in rhesus monkey primates. METHOD: IFN-alpha2b was loaded with biodegradable PLGA with 3 inherent viscosities using a double emulsion and solvent evaporation method. The particle size, surface morphology, and in vitro release profiles were investigated. Two groups of rhesus monkeys (n=3) were injected intramuscularly with either 3 MIU/kg commercial IFN-alpha2b lyophilized powder or IFN-alpha2b-loaded PLGA microspheres (inherent viscosity of 0.89 dL/g). In vitro release was determined by Lowry protein assay. The serum IFN and neopterin levels were determined by the enzyme-linked immunosorbent assay (ELISA) method to evaluate biological activity of the microspheres in rhesus monkeys. RESULTS: The IFN-alpha2b microspheres with 3 inherent viscosities (0.39, 0.89, and 1.13 dL/g) were entirely spherical and had a smooth surface. The average diameter of each type was 45.55, 81.23, and 110.25 microm, respectively. The in vitro release was 30 d. The pharmacokinetic-pharmacodynamic properties between the IFN-alpha2b microspheres and IFN-alpha2b lyophilized powder were significantly different (P<0.05). CONCLUSION: The drug residence time for the IFN-alpha2b of the PLGA microsphere with an inherent viscosity of 0.89 dL/g in plasma significantly increased and had a longer time of biological effects in rhesus monkeys following intramuscular administration.

[Study on chemical constituents of volatile oil and trace elements from fruits of Clausena lansium].

OBJECTIVE: To study and determine the chemical constituents of the volatile oil and the trace elements in the fruits of the Clausena lansium. METHOD: The essential oils were extracted by supercritical fluid extraction (SFE) and separated on capillary columns with HP6890GC-5973MS. The components were quantitatively determined with normalization method, and were identified with GC-MS. And the trace elements were determined by ICP-MS and ICP-AES. RESULT: 36 Components constituting 95% of the total essential oil were separated and identified, and 11 trace elements were identified. CONCLUSION: 18 Compounds were found from the fruit of C. lansium for the first time.

Recurrence of lattice corneal dystrophy caused by incomplete removal of stroma after deep lamellar keratoplasty.

PURPOSE: To report clinical and histopathological characteristics of postoperative amyloidosis recurrence in a patient with lattice corneal dystrophy (LCD) type I. METHODS: The clinical manifestation of recurrent amyloidosis in the residual stroma was delineated in a patient with LCD type I after deep lamellar keratoplasty (DLKP) for 6.5 years. Complete removal of the residual recipient stroma and regrafting of a new cryopreserved donor button were accomplished by a secondary DLKP. The primary DLKP donor graft and the underlying residual stroma of the recipient obtained by the secondary DLKP were examined for analysis of histopathologic and ultrastructural changes. RESULTS: A tongue-shaped retained stroma with linear opacity was observed underneath the primary DLKP donor graft. The retained stromal layer was thoroughly detached from Descemet membrane, removed, and followed by grafting a new cryopreserved button. The primary donor button exhibited a normal epithelium, fewer keratocytes, an intact Descemet membrane, and mild positive Congo red staining in the middle layer of the stroma. The total retained recipient stroma removed by the secondary DLKP measured approximately 20 mum in thickness, showing thick and massive amyloid accumulation. The surface of the removed residual stroma toward Descemet membrane showed collagen fibers in an interwoven fashion without bundle structure under a scanning electron microscope. CONCLUSION: Incomplete removal of the recipient stroma by DLKP can lead to the recurrence of amyloidosis in the residual stroma in patients with LCD. Clinical and histologic findings in the primary graft and in the residual recipient stroma implicate stromal genesis of recurrence of LCD after DLKP.

Construction and expression of a single chain antibody mimicing human ovarian cancer antigen CA125.

One concept for immune therapy of cancer involves induction of antigen mimic antibodies to trigger the immune response against tumor cells. Anti-idiotypic antibodies directed against the antigen-binding site of antibodies specific for tumor antigen may functionally and even structurally mimic antigen and induce anti-anti-idiotypic immune response. Monoclonal antibody WJ02 is one of such anti-idiotypic antibodies, which contains internal image of CA125. In order to improve the immunospecificity of mAb WJ02, we constructed a single chain of mAb WJ02 in Vl-linker-Vh orientation. The scFv-WJ02 could be expressed and secreted in the recombinant Pichia pastoris system. The secreted scFv protein with a molecular weight of 30 kD retained the biological activity of mAb WJ02, which was proved by a direct binding assay and inhibition experiment. Our results indicated that the scFv-WJ02 could be used as a possible tool for idiotypic therapy against ovarian cancer, which might enhance the possibility of eliminating nonspecific responses induced by mAb WJ02.

Mitomycin C, amniotic membrane transplantation and limbal conjunctival autograft for treating multirecurrent pterygia with symblepharon and motility restriction.

BACKGROUND: Treatment of eyes with multirecurrent pterygia associated with severe symblepharon and motility restriction is challenging. A combined surgical procedure of intraoperative mitomycin C, amniotic membrane transplantation and conjunctival limbal autograft was applied for treating such eyes. METHODS: Seven eyes of seven patients who had previously undergone an average of four operations for pterygial removal and who manifested recurrent pterygia associated with severe symblepharon and motility restriction were involved in this retrospective study. The surgical procedures involved clearing fibrovascular membrane over the cornea, extensive excision of epibulbar fibrovascular tissue to the bare sclera, application of 0.02% mytomycin C onto the bare sclera for 5 min and transplantation of preserved human amniotic membrane and conjunctival limbal autograft. RESULTS: Postoperatively, all seven eyes showed rapid epithelialization on the corneal surface in 3-5 days and, on the conjunctival surface, in 10-18 days. For a mean follow-up period of 22.4+/-6.1 months, six eyes recovered deep fornices, smooth and stable ocular surface and full ocular motility without recurrence. One eye showed regrowth of fibrovascular tissue and motility restriction but less severe than before surgery. No complication was noted due to mitomycin C. CONCLUSIONS: Combined intraoperative mitomycin C, amniotic membrane graft and limbal conjunctival autograft are successful approaches for treating multirecurrent pterygia with severe symblepharon to restore the ocular surface integrity and prevent recurrence.