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Background: Circulating tumor cells (CTC) play an important role in the screening and prognosis of lung cancer, but the low efficiency and specificity of CTC isolation obviously restrict its clinical application. The purpose of this study is to explore a new and efficient isolation method of CTC in patients with non-small cell lung cancer (NSCLC) in order to achieve the purpose of early diagnosis of NSCLC., Methods: Three kinds of immunolipid magnetic spheres of epidermal growth factor receptor (EGFR), vimentin and folic acid (FA) were prepared by thin film method. After characterization, the sorting scheme of cell line was explored, the optimal sorting scheme of NSCLC CTC was constructed, and its clinical application value was studied., Results: The average capture efficiency of EGFR, Vimentin and FA magnetic spheres used alone and in combination to lung cancer cell lines was 78%, 79%, 82% and 91%, respectively. In 60 patients with lung cancer, using 2 CTC per 7.5 mL blood as cutoff value, the positive rates of EGFR, Vimentin and FA magnets used alone and in combination were 65.0%, 33.3%, 93.3% and 100%, respectively. It was also found that the number of CTC detected by combined use of the three magnetic spheres was correlated with clinical stages (P<0.05)., Conclusions: The combination of three kinds of magnetic spheres can separate EGFR+, Vimentin+, FA+ expressed CTC, which is beneficial to the downstream analysis of CTC correlation. This study provides a new method to improve the efficiency of NSCLC CTC capture, and verifies that the captured CTC counting method can be used in the auxiliary diagnosis of lung cancer.

Objective: To investigate the correlation of seminal plasma homocysteine (Hcy) and its metabolic factors folate (FA) and cobalamin (VB12) with semen quality in obese men., Methods: We randomly selected 83 male patients with idiopathic infertility for this study and, according to the body mass index (BMI), divided them into a normal BMI (n = 28), an overweight (n = 28) and an obesity group (n = 27). We determined the levels of Hcy, FA and VB12 in the seminal plasma by ELISA and analyzed their correlation with the semen parameters of the patients in different groups., Results: Compared with the normal BMI group, the obese males showed significant decreases in the semen volume (［3.23 ± 0.86］ vs ［2.58 ± 0.77］ ml, P < 0.05), total sperm count (［191.35 ± 103.00］ vs ［121.55 ± 88.08］ ×106, P < 0.05), percentage of progressively motile sperm (［52.88 ± 15.58］ % vs ［38.97 ± 16.52］ %, P < 0.05), seminal plasma Hcy (［7.41 ± 1.29］ vs ［6.62 ± 0.85］ μmol/L, P < 0.05), and VB12 (［282.41 ± 30.38］ vs ［230.07 ± 37.75］ pmol/L, P < 0.05). There were no statistically significant differences in the semen parameters between the overweight group and the normal BMI or the obese group (P > 0.05), or in sperm concentration or the FA level among the three groups of patients (P > 0.05). The levels of seminal plasma Hcy and VB12 were correlated positively with the semen volume (r = 0.281 and 0.242, P < 0.05) and total sperm count (r = 0.229 and 0.258, P < 0.05) but negatively with BMI (r = －0.293 and －0.238, P < 0.05). No correlation, however, was found either between sperm concentration and the percentage of progressively motile sperm (P > 0.05) or between the FA level and BMI or semen parameters (P > 0.05)., Conclusions: The levels of seminal plasma Hcy and VB12 are correlated with BMI, semen volume and total sperm count, which suggests that the concentrations of seminal plasma Hcy and VB12 may be associated with the fertility of obese men.

A novel screening method for dihydrofolate reductase inhibitors (DHFRI) using high performance capillary electrophoresis (HPCE) was developed. The separation was performed in a fused silica capillary column using disodium tetraborate (50 mmol/L, pH 9. 18) buffer solution with 0. 002% Brij-35. The separation temperature was controlled at 25 degrees C and a voltage of 25 kV was applied. The detection wavelength was 280 nm. Five microL of various concentrations of inhibitor was added to the enzyme reaction system consisting of 25 micrpL of 50 mmol/L pH 6. 0 potassium phosphate buffer, 10 microL of 0. 025 mg/mL dihydrofolate (FH2), 10 microL of dihydrofolate reductase (DHFR) (0. 25 unit) and 5 microL of 0. 5 mg/mL NADPH in a 300-miL sample tube, then mixed for 1 min and incubated for 30 min at room temperature. The reaction mixture injections were performed by pressure at 3.45 kPa (0. 5 psi) for 10 s. With the developed CE method the substrate and product were baseline resolved within 19 min. The time course of the reaction was studied to show excellent correlation. Quantitative analysis of DHFR inhibition was performed by determining its dynamic parameters. The difference of peak areas between FH2 and FH4 was used to calculate the inhibitory rate. Three inhibitors, amethopterin, trimethoprim and folic acid, were used in the enzyme reaction system to test this method and their IC50 values determined were close to the literature values. It was demonstrated that the developed method is suitable for screening DHFRIs.

Double divisor-ratio spectra derivative method has been developed and applied to determining two ternary mixtures simultaneously by E. Dinc. In fact, E. Dinc's double divisor-ratio spectra derivative method is completely wrong. It can not be applied to determining ternary mixtures simultaneously. This paper proves the Mistake of E. Dinc's method in both theory and practice.

Vitamin B12(VB12), folate and reactive oxygen species(ROS) in human semen from both fertile and infertile males, and their relationships with other parameters were observed. Semen samples from 44 infertile and 176 fentile control subjects were collected and measured based on the guidelines issued by WHO. The results showed that no significant differences on semen VB12 and folate were observed between fertile and infertile groups(P > 0.05). However, the levels of ROS in infertile group were significantly higher than those in fertile group(P < 0.01). The morphological defects and low motility of sperm in infertile group was significantly higher than those in fertile group(P < 0.01). A positive relation was observed between the levels of ROS production and the morphological defect of sperm(P < 0.01). There was a significant negative correlation between the levels of VB12, folate and ROS(P < 0.01) in semen. It is interesting to note that the values of ROS also show a significantly positive correlation with sperm motility (P < 0.01). A significant differences of ROS levels between fertile and infertile group with negative white blood cells in sperm was also observed. It is concluded that the increase of ROS on the morphological defect of sperm is one of the most important factors related to poor sperm quality and human infertility.

In order to measure the content of folate in foods accurately, to hydrolyze and extract folate in foods with suitable pretreatment is necessary because folate in foods is always in polyglutamate structure. The pretreatment of samples including hydrolysis with phosphate buffer(0.05 mol/L, pH 6.8) once or twice, with chick pancreatin, protease and/or amylase, and a combining of trienzyme methods was compared and discussed. A microbiological method with lactobacillus casei was used. A high result of folate in foods could be obtained when extracting folate enriched samples or vegetables with phosphate buffer once or twice respectively, or hydrolyzing "extracted oral liquids" with chick pancreatin, or when extracting samples rich in protein and/or starch with trienzyme. The RSD of interassay and intrassay were 6.5%-9.9% and 3.5%-14.1% respectively, and the recovery rate was 86.8%-106.8%.