OBJECTIVE: To extract sperm DNA from mixed stain by the modified differential lysis method combined with silicon bead method and to evaluate its application value. METHODS: Fifty-two mixed stains containing female STR genotypes detected by differential lysis method were collected. The sperm DNA was extracted by the modified method combined with silicon bead method, then genotyped with the Identifiler Kit, and compared with the results of genotyping by the conventional differential lysis method as control. RESULTS: Of the 52 samples, 38 samples with sole male STR genotypes in all loci were detected. The detection rate of male STR genotypes was 98.08% through the modified method combined with silicon bead method. CONCLUSION The modified differential lysis method combined with silicon bead method can be used in extraction of sperm DNA from mixed stain.
DNA/isolation & purification* ; DNA Fingerprinting ; Female ; Genotype ; Humans ; Male ; Polymerase Chain Reaction ; Silicon ; Spermatozoa

The protoplasts of original Aspergillus niger strain Uco-3 were treated with the cooperation of UV and ?-ray to obtain the high-yielding strain producing the thermostable ?-galactosidase. Under the optimum conditions of formation and regeneration protoplasts were prepared. According to the interaction of positive mutation rate and radiation dose,the optimum condition was determined. The optimum dose of UV was 4 minutes and the optimum dose of ?-ray was 500 Gy. After mutagenetic treatment of protoplasts and selection from a lot of mutants,a mutant DL116 producing the thermostable ?-galactosidase was obtained. The ?-galactosidase activity of DL116 was increased from 16.27 U/mL to 44.37 U/mL,which was higher than that of strain Uco-3.

A novel Aspergillus terreus strain M11 was isolated from the compost containing cellulose and identified. The isolate grow best at 45℃ and pH2.0. It was found that the activity of the CMCase was up to 3.680IU/mL with high heat stability and the optimal reaction conditions of the CMCase were at 60℃ and pH2.0.

A novel drug delivery system combining oral fast dissolving film with sodium deoxycholate/phospholipid mixed micelles was prepared to increase the absorption of cucurbitacin B that is a poor aqueous solubility substance. Encapsulation efficiency, particle size, zeta potential, polydispersity coefficient, investigated the morphology, disintegration time of oral fast dissolving film and the pharmacodynamic properties of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles before and after solidified in mice were evaluated and compared. The oral fast dissolving film prepared in this study showed a homogeneous pale yellow and could completely disintegrated in the 30 s. It could meet the requirements of rapidly disintegrating fully. The encapsulation efficiency, particle size, zeta potential, polydispersity coefficient of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles loaded in oral fast dissolving film were (43.36 +/- 2.12)%, (108.82 +/- 5.2) nm, (-34.18 +/- 1.07) mV, 0.088 +/- 0.012, respectively. The encapsulation efficiency, particle size, zeta potential, polydispersity coefficient of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles in solution were (41.26 +/- 2.22)%, (181.82 +/- 4.48) nm, (-30.67 +/- 0.81) mV, 0.092 +/- 0.012, respectively. The difference of pharmacodynamics among film of cucurbitacin B-loaded micelles, cucurbitacin B-loaded micelles and free cucurbitacin B in vivo was compared. Solubility of cucurbitacin B loaded in sodium deoxycholate/phospholipid-mixed micelles has also been greatly improved. The tumor inhibition rate of cucurbitacin B loaded in sodium deoxycholate/phospholipid-mixed micelles was significantly improved and did not change significantly before and after solidified. These showed that the sodium deoxycholate/phospholipid-mixed micelles could enhance the antitumor activities of cucurbitacin B and the stability of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles was improved significantly after solidified by oral fast dissolving film technology without pharmacodynamic properties changed significantly.
Animals ; Antineoplastic Agents ; administration & dosage ; chemistry ; Cell Line, Tumor ; Deoxycholic Acid ; chemistry ; Drug Carriers ; chemistry ; Humans ; Male ; Mice ; Neoplasms ; drug therapy ; Phospholipids ; chemistry ; Solubility ; Triterpenes ; administration & dosage ; chemistry

Objective To assessed the expression of inducible costimulator(ICOS)on peripheral blood and joint fluid CD4,CDS,CD45RO T cells and B cells in rheumatoid arthritis(RA).Methods Expression of ICOS and ICOS/CD45RO on peripheral blood and joint fluid CD4~+CD8~+T cells and ICOS ligand(ICOSL)on CD19 B cells from RA patients and healthy volunteers were determind by three-color flow cytometry.Compar- ision with active and inactive RA,initial and relapsed RA had been done.Results Joint fluid CD4 and CD8 T cells expressing ICOS,ICOS/CD45RO were significantly increased than peripheral blood in RA patients and healthy subjects.Joint fluid B cells expressing ICOSL were significantly reduced than peripheral blood in RA patients.Meanwhile,peripheral blood B cells expressing ICOSL were significantly reduced in active RA than inactive RA patients.Conclusion Hyperexpression of ICOS and ICOS/CD45RO on joint fluid CD4 and CD8 T cells and lowexpression of ICOSL in B cells from RA patients,expecially in active RA may contribute to the local immunopathological roles and joint destructions in the pathogenesis of RA.

The influence of low tube voltage in dual source CT (DSCT) coronary artery imaging on image quality and radiation dose and its application value in clinical practice were investigated. Totally, 300 cases of chest pain with low body mass index (BMI <18.5 kg/m(2)) subjected to DSCT coronary artery imaging were prospectively enrolled. The heart rate in all patients were greater than 65/min. The retrospective ECG gated scanning mode and simple random sampling method were used to assign the patients into groups A, B and C (n=100 each). The patients in groups A, B and C experienced 120-, 100-, and 80-kV tube voltage imaging respectively, and the image quality was evaluated. The CT volume dose index (CTDIvol) and dose length product (DLP) were recorded, and the effective dose (ED) was calculated in each group. The image quality scores and radiation doses in groups were compared, and the influence of tube voltage on image quality and radiation dose was analyzed. The results showed that the excellent rate of image quality in groups A, B and C was 95.69%, 94.72% and 96.33% respectively with the difference being not statistically significant among the three groups (P>0.05). The CTDIvol values in groups A, B and C were 51.35±12.21, 21.28±7.13 and 6.34±3.34 mGy, respectively, with the difference being statistically significant (P<0.05). The ED values in groups A, B and C were 9.27±1.63, 4.56±2.29 and 2.29±1.69 mSv, respectively, with the difference being statistically significant (P<0.05). It was suggested that for the patients with low BMI, the application of DSCT coronary artery imaging with low tube voltage can obtain satisfactory image quality, and simultaneously, significantly reduce the radiation dose.

Objective: To summarize the influence patterns of related time factors on acupuncture effectiveness in different disease model rats/mice, and to provide reference for acupuncture clinical practice. Methods: Retrieved the relevant literatures on time-effect experimental studies of acupuncture in rats/mice in the recent 10 years. The correlations between the key time factors (such as different intervention timings of acupuncture, acupuncture moments, operation durations, needle-retaining times, intervals and treatment courses) and the acupuncture effect were analyzed and summarized. Results: From the mainstream perspective of quantification, the earlier the acupuncture intervention, the better. The proper time to implement acupuncture varied depending on disease models and points. The best operation time varied widely between different diseases. The most frequently needle-retaining time was 20-30 min. The frequency of acupuncture was usually 1 time/day; the length of the treatment course was determined according to practitioners' experience. Conclusion: Throughout the time-effect studies of acupuncture intervention in experimental rats/mice, conclusions are inconsistent, especially the lack of quantitative research on acupuncture operation time, acupuncture frequency, acupuncture treatment duration, and optimal stimulation amount. Future research should explore and determine the best time-quantity parameters that affect the effectiveness of acupuncture intervention, which is the key and goal of the acupuncture time-effect research. Independent intervention-time experiments throughout the entire course of a single disease (dominant disease) need to be done to guide clinical and disciplinary development.

OBJECTIVETo compare the effects and complications of subinguinal microscopic ligation, laparoscopic transperitoneal varicocelectomy, laparoscopic retroperitoneal varicocelectomy, open retroperitoneal high ligation, and interventional embolotherapy in the treatment of varicocele.

METHODSWe conducted a retrospective study that included 632 varicocele patients treated by subinguinal microscopic ligation (group A, n = 79), laparoscopic transperitoneal varicocelectomy (group B, n = 120), laparoscopic retroperitoneal varicocelectomy (group C, n =137), open retroperitoneal high ligation (group D, n = 283), and interventional embolotherapy (group E, n = 13). We compared the baseline and 3-month postoperative semen parameters, postoperative complications, and pregnancy rate among the five groups of patients.

RESULTSThe operation time was longer in groups A ([2.02 ± 1.25] h) and E ([2.17 ± 1.02] h) than in the other three groups, while the postoperative hospital stay was the shortest in group E ([1.1 ± 0.1] d). Intestinal injury or incision bleeding occurred intraoperatively in 2 cases in group B and 1 case in group E. Postoperative scrotal edema developed in 3.7, 17, 10, and 19% of the patients in groups A, B, C, and D, respectively, but not in group E. The rate of 1-year recurrence was the lowest in group A (1.6%) and highest in group E (22%). Sperm concentration and the percentages of progressively motile sperm and morphologically normal sperm were improved postoperatively in all the patients (P < 0. 05), but there were no statistically significant differences among the five groups either in the above three parameters or in the postoperative pregnancy rate (P > 0. 05).

CONCLUSIONIn the surgical treatment of varicocele, laparoscopic retroperitoneal approach involves short operation time and few complications, subinguinal microscopic ligation has the advantages of little injury, rapid recovery, and few complications but requires specialized microsurgical techniques, and interventional embolotherapy leaves no incision scar and needs only local anesthesia and 1-day postoperative hospital stay, which is uitable for those with a contraindication to anesthesia.

Embolization, Therapeutic ; adverse effects ; methods ; Female ; Humans ; Laparoscopy ; adverse effects ; methods ; Length of Stay ; Ligation ; adverse effects ; methods ; Male ; Operative Time ; Postoperative Complications ; Postoperative Hemorrhage ; Pregnancy ; Pregnancy Rate ; Recurrence ; Retroperitoneal Space ; Retrospective Studies ; Sperm Count ; Urogenital Surgical Procedures ; adverse effects ; Varicocele ; surgery ; Vascular Surgical Procedures ; adverse effects ; methods

OBJECTIVETo study the impact of left varicocele (VC) and varicocelectomy (VCT) on the apoptosis of spermatogenic cells and the levels of nitrogen monoxidum (NO) and interleukin 1 (IL-1) in the rat testis.

METHODSWe randomly divided 60 adolescent male SD rats into four groups of equal number: sham operation control, VC model 1 (VC1), VC model 2 (VC2), and VCT. We determined the semen quality and levels of NO and IL-1 in the testis tissue, detected the apoptosis of spermatogenic cells by TUNEL, and compared the indexes obtained among different groups.

RESULTSAn experimental VC model was successfully established by partially ligating the left renal vein of the rats. Sperm concentration and motility were significantly decreased in the VC1 ([1.54 ± 1.16] x 10⁶/ml and [44.23 ± 15.46]%) as compared with those in the sham operation group ([2.80 ± 1.62] x 10⁶/ml and [72.34 ± 12.62]%) (P < 0.05), but remarkably higher in the VCT ([1.82 ± 1.34] x 10⁶/mI and [51.21 ± 12.62]%) than in the VC2 group ([1.04 ± 1.21] x 10⁶/ml and [39.23 ± 13.21]%) (P < 0.05). The levels of NO and IL-1 in the left testes were markedly elevated in the VC1 ([0.172 ± 0.030] ng/ml and [1.468 ± 0.080 ] mg/ml) in comparison with those in the sham operation group ([0.134 ± 0.021] ng/ml and [0.782 ± 0.079 ] mg/ml) (P < 0.05), and significantly higher in the VC2 ([0.198 ± 0.020] ng/ml and [1.994 ± 0.090] mg/ml) than in the VCT group ([0.141 ± 0.010] ng/ml and [0.781 ± 0.036] mg/ml) (P < 0.05). However, the NO and IL-1 levels in the right testis showed no statistically significant differences between the two groups, and the two levels were positively correlated (r = 0.492, P < 0.01). The rats of the VC1 group exhibited remarkable apoptosis of spermatogenic cells in the bilateral testes, with significant differences in the apoptosis index ( AL) between the two sides (P < 0.05) as well as in the same side in comparison with the sham operation group (P < 0.01). The Als of spermatogenic cells in the bilateral testes showed statistically significant differences in the VCT (P < 0.05) but not in the VC2 group (P > 0.05), and those in the same side manifested dramatic differences between the VCT and VC2 groups (P < 0.01).

CONCLUSIONVaricocele induces changes of the NO and IL-1 levels in the testis tissue and increases the apoptosis of spermatogenic cells, which might be one of the causes of testis damage and spermatogenic dysfunction.

Animals ; Apoptosis ; Germ Cells ; pathology ; In Situ Nick-End Labeling ; Interleukin-1 ; analysis ; Ligation ; Male ; Nitrogen ; analysis ; Random Allocation ; Rats ; Semen Analysis ; Spermatogenesis ; Testis ; chemistry ; Varicocele ; complications ; surgery

Cinobufacino injection is purified from water extraction of the skin of Bufo bufo gargarizans, which has been widely used for various cancers in clinic with significant anti-tumor effects. Bufadienolides were regarded as the main active constituents of cinobufacino injection in previous reports. In present study, 6 bufadienolides were isolated and purified from Cinobufacino injection. Their structures were identified as 3-epi-ψ-bufarenogin (1), ψ-bufarenogin (2), 3-epi-arenobufagin (3), arenobufagin (4), 3-epi-gamabufotalin (5), and 3-oxo-arenobufagin (6), separately. Among them, 1 and 3 were new compounds, 5 and 6 were new natural products. Compounds 1, 2 and compounds 3, 4 were two pairs configuration isomers at C-3, separately.
Animals ; Bufanolides ; chemistry ; isolation & purification ; Bufo bufo ; Injections ; Skin ; chemistry