[Efficacy comparison of zero-profile intervertebral fusion and stand-alone interbody cage combined with cage-titanium plate construct in treatment of two-segment skip cervical spondylosis].

Objective: To investigate the difference in clinical efficacy between zero-profile interbody fusion (ROI-C) and stand-alone interbody cage combined with cage-titanium plate construct for patient with two-segment skipped cervical spondylosis who received the anterior cervical discectomy and fusion (ACDF) surgical strategies. Methods: The clinical data of 62 patients with two-segment skipped cervical spondylosis who underwent surgical treatment in the First Affiliated Hospital of Zhengzhou University from June 2017 to June 2020 were retrospectively analyzed, included 38 males and 24 females, aged (53.3±8.5) years. Thirty-three cases were treated with ROI-C (ROI-C group), and 29 cases with stand-alone interbody cage combined with cage-titanium plate construct (cage+titanium plate group). The following parameters, including operation time, intraoperative blood loss, dysphagia Bazaz grade, Japanese Orthopaedic Association (JOA) score, visual analogue scale (VAS) of pain, neck disability index (NDI), average intervertebral height of operated segments, C2-7 Cobb angle, and related complications, were compared between the two groups. Results: The patients were followed up for (28±5) months (16-34 months). The operation time of ROI-C group was (127.6±34.2) min, which was shorter than that in cage+titanium plate group [(157.1±43.9) min, P=0.004]. The scores of JOA and VAS in both ROI-C group and cage+titanium plate group were significantly improved 3 months after operation when compared with those before operation, and there was no significant difference between the two groups (all P>0.05). The average intervertebral space height of fusion segment in ROI-C group was (6.02±1.03) mm before operation, it was improved to (8.38±1.47) mm at 3 months after operation, (8.16±1.40) mm at 12 months after operation, and it was (6.24±1.05) mm, (8.58±1.18) mm and (7.87±0.73) mm in cage+titanium plate group, respectively, and there was no significant difference between the two groups at each time point (all P>0.05). The Cobb angle of cervical vertebrae in ROI-C group was 10.5°±6.8° before operation, improved to 19.2°±9.0° at 3 months after operation, 18.2°±5.8° at 12 months after operation, and it was 10.9°±4.6°, 18.5°±7.8°, 17.1°±5.2° in cage+titanium plate group, respectively, and there was no significant difference between the two groups at each time point (all P>0.05). The incidence of postoperative dysphagia was 9.1%(3/33) in the ROI-C group and 37.9%(11/29) in the cage+titanium plate group, and the difference was statistically significant (P=0.007). Conclusions: Both ROI-C and stand-alone interbody cage combined with cage-titanium plate construct can achieve good results for two-segment skipped cervical spondylosis. However, ROI-C is more advantageous in shortening the operation time and reducing early postoperative dysphagia.

An odorant receptor mediates the attractiveness of cis-jasmone to Campoletis chlorideae, the endoparasitoid of Helicoverpa armigera.

Parasitic wasps rely on olfaction to locate their hosts in complex chemical environments. Odorant receptors (ORs) function together with well-conserved odorant coreceptors (ORcos) to determine the sensitivity and specificity of olfactory reception. Campoletis chlorideae (Hymenoptera: Ichneunmonidae) is a solitary larval endoparasitoid of the cotton bollworm, Helicoverpa armigera, and some other noctuid species. To understand the molecular basis of C. chlorideae's olfactory reception, we sequenced the transcriptome of adult male and female heads (including antennae) and identified 211 OR transcripts, with 95 being putatively full length. The tissue expression profiles, as assessed by reverse-transcription PCR, showed that seven ORs were expressed only or more highly in female antennae. Their functions were analysed using the Xenopu slaevis oocyte expression system and two-electrode voltage-clamp recordings. CchlOR62 was tuned to cis-jasmone, which was attractive to female C. chlorideae adults and H. armigera larvae in the subsequent behavioural assays. Further bioassays using caged plants showed that the parasitism rate of H. armigera larvae by C. chlorideae on cis-jasmone-treated tobacco plants was higher than on the control plants. Thus, cis-jasmone appears to be an important infochemical involved in the interactions of plants, H. armigera and C. chlorideae, and CchlOR62 mediates the attractiveness of cis-jasmone to C. chlorideae.

Combination of mRNA of Repair-related Genes in Rat Skeletal Muscles for Wound Age Estimation.

ABSTRACT: Objective To investigate the estimation of early and mid-term wound age by a combination of four mRNAs, the DNA polymerase delta-interacting protein 3 （POLDIP3） mRNA, regulator of chromosome condensation 1 like （RCC1L） mRNA, proline-rich 5 （PRR5） mRNA, and ribonucleic acid export 1 （RAE1） mRNA in rats skeletal muscles. Methods The model of rat skeletal muscle contusion was established, and then contusion area muscle tissue was extracted 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44 and 48 h after injury. Histomorphological changes during the repair process after rat skeletal muscle contusion were observed. The relative expressions of Poldip3, Rcc1l, Prr5 and Rae1 mRNAs were detected by reverse transcription real-time quantitative polymerase chain reaction （RT-qPCR）. Different stages of wound age were classified by using the expression patterns of four genes at various time points after injury. The accuracy of the results was verified by Fisher discriminant analysis. Results Histomorphological results showed that the repair process after skeletal muscle contusion occurred with the prolonging of time. Through combination of the expression trends of the four kinds of mRNAs, the 48 h after injury could be divided into three periods, 4-12 h, 16-28 h and 32-48 h. The Fisher discrimination method showed that the classification accuracy rates of the three periods were 83.3%, 75.0% and 73.3%, respectively. Conclusion The classification discrimination based on the relative expression of every gene has a higher accuracy, and the accuracy of wound age estimation with combination of mRNA relative expressions is higher than that with a single indicator. By combining with Fisher discrimination method, this method can be used for early and mid-term wound age estimation.

Resonant structures for infrared detection.

We are developing resonator-quantum well infrared photodetectors (R-QWIPs) for long-wavelength applications. Detector pixels with 25 µm pitch were hybridized to fan-out circuits for radiometric measurements. With a moderate doping of 0.5×1018 cm-3, we achieved a quantum efficiency (QE) of 37% and conversion efficiency (CE) of 15% in a 1.3 µm thick active material and 35% QE and 21% CE in a 0.6 µm thick active material. Both detectors are cutoff at 10.5 µm with a 2 µm bandwidth. The temperature at which photocurrent equals dark current is about 65 K under F/2 optics. The thicker detector shows a large QE polarity asymmetry due to nonlinear potential drop in the QWIP material layers.

Intestinal absorption mechanisms of ginsenoside Rh2: stereoselectivity and involvement of ABC transporters.

This study investigated the absorption mechanism of ginsenoside Rh2 to clarify the reasons for its poor absorption. Transepithelial transport across Caco-2 cell monolayers, cellular uptake, and in situ rat intestinal perfusion were examined. Cellular uptake of Rh2 was linear from 1 to 50 µM at 4°C, whereas it was saturated when the concentration exceeded 10 µM at 37°C. At 37°C, the uptake at 10 µM was linear in 60 min. Intracellular exposure in 240 min was 2173.70 and 979.38 ng·min/µg for S and R isomers, respectively. Transepithelial permeability of Rh2 was about 10⁻8 to 10⁻7 cm/s. Efflux ratios were above 1.5. Sodium dodecyl sulfate, sodium citrate, and sodium deoxycholate had no effect on Rh2 permeability. After intestinal perfusion for 3 h, 9.1% of 20(R)-Rh2 and 15.7% of 20(S)-Rh2 were absorbed. Cyclosporine, quercetin, and probenecid could improve the cellular uptake, absorptive permeability, and intestinal absorption. Carrier-mediated transport was the major absorption mechanism. Rh2 was a substrate of ABC transporters. The ABC-transporter-mediated efflux and the poor permeability were the major reasons for Rh2 poor absorption. The stereoselective absorption was significant. R isomer exhibited lower absorption profiles in all the experiments, possibly due to more potent efflux.

[Short-term efficacy evaluation of laparoscopic spleen-preserving splenic hilus lymphadenectomy and left epigastrium mesogastric excision for advanced proximal gastric cancer based on mesangial anatomy].

Objective: To investigate short-term efficacy of laparoscopic spleen-preserving splenic hilus lymphadenectomy and left epigastrium mesogastric excision for advanced proximal gastric cancer based on mesangial anatomy. Methods: A case series study was carried out. Case inclusion criteria: (1) patient was confirmed as gastric adenocarcinoma by gastroscopic biopsy before operation; (2) locally advanced gastric cancer was confirmed by abdominal CT before operation; (3) no distant metastases such as liver, lung, and posterior peritoneal lymph nodes, and no tumor directly invading the pancreas, spleen, liver, and colon were verified by superficial lymph node ultrasound, chest and abdominal CT before operation;(4) total gastrectomy or proximal gastrectomy plus D2 lymphadenectomy were performed, and R0 resection was confirmed by postoperative pathology. Exclusion criteria: (1) intraperitoneal dissemination or distant metastasis was found during laparoscopic exploration; (2) No.10 lymph nodes were significantly enlarged or fused into clusters; (3) pathological diagnostic data were incomplete. According to above criteria, the clinicopathological data of 36 patients who underwent laparoscopic spleen-preserving No.10 lymphadenectomy and left epigastrium mesogastric excision based on interspace anatomy for advanced proximal gastric cancer in The First Affiliated Hospital of Zhengzhou University from June 2017 to March 2018 were retrospectively collected and analyzed. The intraoperative conditions, postoperative recovery and complications of patients were analyzed. Results: In 36 patients, the mean age was (59.8±8.0) years, the mean BMI was (23.9±3.5) kg/m(2), and 8 cases (22.2%) received preoperative chemotherapy. All the patients underwent successfully the laparoscopic spleen-preserving splenic hilus lymphadenectomy and left epigastrium mesogastric excision. In the examination of postoperative resected specimens, it was found that the mesangial boundary of the upper and posterior part of the stomach was smooth, indicating the efficiency of complete mesangial resection. No case was converted to open operation. The mean time of lymph node dissection and mesangial resection was (34.2±11.4) minutes. The mean blood loss during operation was (44.8±21.3) ml. The mean number of lymph node dissection per patient was 45.6±17.6. The mean number of No. 11p+11d lymph node dissection was 3.1± 2.8 per patient, and 7 patients were pathologically positive with metastasis rate of 19.4% (7/36). The mean number of No.10 lymph node dissection was 2.9±2.5 per patient, and 2 patients were pathologically positive with metastasis rate of 5.6% (2/36). The time to postoperative flatus was (3.8±0.6) days, time to removal of nasogastric was (1.9±0.7) days, time to the first intake of fluid was (3.0±0.4) days, time to removal of drainage tube was (6.0±1.2) days. Postoperative mean hospital stay was (12.8±4.0) days. One case (2.7%) developed pulmonary embolism and 1 case (2.7%) developed gastroplegia after operation. The morbidity of postoperative complication was 5.6% (2/36). No operative site infection, postoperative bleeding and death within postoperative 30-day were observed. All the 36 patients were followed up and the median follow-up was 18 months (12-28 months). Seven patients died of tumor relapse and metastasis (3 cases died within postoperative 1 year) and another 1 case developed colonic cancer 17 months after operation. Conclusion: Laparoscopic spleen-preserving splenic hilus lymphadenectomy and left epigastrium mesogastric excision for advanced proximal gastric cancer based on mesangial anatomy is safe and feasible.

[Clinical application effect of modified nasopharyngeal swab sampling for 2019 novel coronavirus nucleic acid detection].

Objective: To study the clinical application effect of modified nasopharyngeal swab sampling for 2019 novel coronavirus nucleic acid detection. Methods: This study covered the period from January 14 to March 1, 2020.The supine position method and the protective face screen were used to collect nasopharyngeal swabs from February 24 onwards, before which, the nasopharyngeal swabs were collected by sitting position method. All the patients who were diagnosed with suspected/confirmed 2019 novel coronavirus infection were admitted from February 19 with the nasopharyngeal swabs collected outside the hospital before admission. (1) Thirty-four swabbing operators meeting the inclusion criteria of the study were recruited in this retrospective cohort study. They were grouped according to the collection method of nasopharyngeal swabs. Sixteen operators of Wuhan Taikang Tongji Hospital who applied the supine position method and the protective face screen were included in supine position method+ protective face screen group (15 males and 1 female, aged 34-49 years); 18 operators (12 from the First Affiliated Hospital of Army Medical University (the Third Military Medical University), 1 from Wuhan Jiangxia Mobile Cabin Hospital, 5 from the East District of People's Hospital of Wuhan University) who applied the traditional sitting position method were included in sitting position method group (2 males and 16 females, aged 25-49 years). In supine position method+ protective face screen group, when collecting sample, the patient lay flat and wore a special protective face screen for nasopharyngeal swab sampling, with neck slightly extending and face turning to the opposite side of the operator about 10°. The self-designed questionnaire was used to investigate the cooperation, the incidence of nausea, coughing, sneezing, and struggling of patients evaluated by the operators, the operation time for a single swab sample, the fear of operation and the perceived exposure risk of operators in the two groups. (2) Sixty-five patients (22 males and 43 females, aged 25-91 years) admitted to Wuhan Taikang Tongji Hospital who successively received the sitting position method and supine position method+ protective face screen for nasopharyngeal swabs sampling and with complete nucleic acid detection results were included. The positive rates of nucleic acid detection by the two sampling methods of nasopharyngeal swabs of the patients were statistically analyzed. (3) Forty-one patients who could express their feelings accurately were selected out of those 65 patients (12 males and 29 females, aged 27-83 years). The comfort of patients in the process of sampling by the two methods was investigated. (4) Thirty-four patients (10 males and 24 females, aged 25-83 years) with two or more consecutive negative results of nucleic acid detection of nasopharyngeal swabs by sitting position method were selected from the above 65 patients. The positive rate of nucleic acid detection of nasopharyngeal swab of patients by supine position method+ protective face screen, i. e. negative to positive rate was statistically analyzed. Data were statistically analyzed with t test, Wilcoxon signed rank test, and chi-square test. Results: (1) Compared with those of sitting position method group, the cooperation score of patients evaluated by the operators in supine position method+ protective face screen group was significantly higher (Z=-4.928, P<0.01), the incidence of nausea, choking cough, sneezing, and struggling of patients evaluated by the operators, and the fear of operation score and the perceived exposure risk score of operators in supine position method+ protective face screen group were significantly lower (Z=-5.071, -5.046, -4.095, -4.397, -4.174, -5.049, P<0.01), and the operation time for a single swab sample in supine position method+ protective face screen group was significantly longer (t=223.17, P<0.01). (2) The positive rate of nucleic acid detection of nasopharyngeal swabs by supine position method+ protective face screen was 60.00% (39/65), which was obviously higher than 41.54% (27/65) by sitting position method (χ(2)=4.432, P<0.05). (3) The comfort score of the 41 patients during nasopharyngeal swabs sampling by supine position method+ protective face screen was significantly higher than that by sitting position method (Z=-5.319, P<0.01). (4) Of the 34 patients with two or more consecutive negative results of nucleic acid detection of nasopharyngeal swabs by sitting position method, the rate of negative to positive of nucleic acid detection was 26.47% (9/34) after sampling by supine position method+ protective face screen. Conclusions: Compared with the traditional sitting position method, detection of 2019 novel coronavirus nucleic acids of nasopharyngeal swabs collected by supine method combined with protective face screen is worth promoting, because of its better comfort of patients, low exposure risk for operators, in addition to reducing in the false negative result to some extent, which may help reduce false recurrence of discharged patients.

Thickness measurement by two-sided step-heating thermal imaging.

Infrared thermal imaging is a promising nondestructive technique for thickness prediction. However, it is usually thought to be only appropriate for testing the thickness of thin objects or near-surface structures. In this study, we present a new two-sided step-heating thermal imaging method which employed a low-cost portable halogen lamp as the heating source and verified it with two stainless steel step wedges with thicknesses ranging from 5 mm to 24 mm. We first derived the one-dimensional step-heating thermography theory with the consideration of warm-up time of the lamp, and then applied the nonlinear regression method to fit the experimental data by the derived function to determine the thickness. After evaluating the reliability and accuracy of the experimental results, we concluded that this method is capable of testing thick objects. In addition, we provided the criterions for both the required data length and the applicable thickness range of the testing material. It is evident that this method will broaden the thermal imaging application for thickness measurement.

Simultaneous measurement of thermal conductivity and heat capacity by flash thermal imaging methods.

Thermal properties are important for material applications involved with temperature. Although many measurement methods are available, they may not be convenient to use or have not been demonstrated suitable for testing of a wide range of materials. To address this issue, we developed a new method for the nondestructive measurement of the thermal effusivity of bulk materials with uniform property. This method is based on the pulsed thermal imaging-multilayer analysis (PTI-MLA) method that has been commonly used for testing of coating materials. Because the test sample for PTI-MLA has to be in a two-layer configuration, we have found a commonly used commercial tape to construct such test samples with the tape as the first-layer material and the bulk material as the substrate. This method was evaluated for testing of six selected solid materials with a wide range of thermal properties covering most engineering materials. To determine both thermal conductivity and heat capacity, we also measured the thermal diffusivity of these six materials by the well-established flash method using the same experimental instruments with a different system setup. This paper provides a description of these methods, presents detailed experimental tests and data analyses, and discusses measurement results and their comparison with literature values.

Absorption enhancement study of astragaloside IV based on its transport mechanism in caco-2 cells.

The purpose of this study was to investigate the transport characteristics and mechanisms for discovering the possible causes of the low bioavailability of astragaloside IV and to develop an absorption enhancement strategy. Caco-2 cells used as the in vitro model. Results showed a low permeability coefficient (3.7 x 10(-8)cm/s for transport from the AP to BL direction), which remained unchanged throughout the concentration range studied, indicating that the transport of astragaloside IV was predominantly via a passive route. The AP to BL transport of astragaloside IV was found to be highly sensitive to the extracellular Ca2+ concentration, which suggested that its transport may be via a paracellular route. Both chitosan and sodium deoxycholate can increase the permeation efficiency of astragaloside IV. This study indicated that astragaloside IV having a low fraction dose absorbed in humans mainly due to its poor intestinal permeability, high molecular weight, low lipophilicity as well as its paracelluar transport may directly result in the low permeability through its passive transport. Meanwhile, chitosan and sodium deoxycholate can be used as absorption enhancers based on its transport mechanism.

Differential mercury volatilization by tobacco organs expressing a modified bacterial merA gene.

Mercury pollution is a major environmental problem accompanying industrial activities. Most of the mercury released ends up and retained in the soil as complexes of the toxic ionic mercury (Hg2+), which then can be converted by microbes into the even more toxic methylmercury which tends to bioaccumulate. Mercury detoxification of the soil can also occur by microbes converting the ionic mercury into the least toxic metallic mercury (Hg0) form, which then evaporates. The remediation potential of transgenic plants carrying the MerA gene from E. coli encoding mercuric ion reductase could be evaluated. A modified version of the gene, optimized for plant codon preferences (merApe9, Rugh et al. 1996), was introduced into tobacco by Agrobacterium-mediated leaf disk transformation. Transgenic seeds were resistant to HgCl2 at 50 microM, and some of them (10-20% ) could germinate on media containing as much as 350 microM HgCl2, while the control plants were fully inhibited or died on 50 microM HgCl2. The rate of elemental mercury evolution from Hg2+ (added as HgCl2) was 5-8 times higher for transgenic plants than the control. Mercury volatilization by isolated organs standardized for fresh weight was higher (up to 5 times) in the roots than in shoots or the leaves. The data suggest that it is the root system of the transgenic plants that volatilizes most of the reduced mercury (Hg0). It also suggests that much of the mercury need not enter the vascular system to be transported to the leaves for volatilization. Transgenic plants with the merApe9 gene may be used to mercury detoxification for environmental improvement in mercury-contaminated regions more efficiently than it had been predicted based on data on volatilization of whole plants via the upper parts only (Rugh et al. 1996).

Sperm deoxyribonucleic acid fragmentation is increased in poor-quality semen samples and correlates with failed fertilization in intracytoplasmic sperm injection.

OBJECTIVE: To determine the incidence of DNA fragmentation in human sperm used for intracytoplasmic sperm injection (ICSI) and to correlate any detected DNA damage with semen analysis parameters and fertilization rates in ICSI. DESIGN: Descriptive and correlational clinical study. SETTING: Tertiary care fertility clinic. PATIENT(S): A total of 150 semen samples was collected from men in the ICSI program. INTERVENTION(S): For each sample, sperm wash and swim-up were performed, and the percentage of recovered sperm with DNA fragmentation was determined with the use of terminal transferase-mediated deoxyuridine triphosphate-biotin end labeling. MAIN OUTCOME MEASURE(S): The percentage of sperm with DNA fragmentation was correlated with semen analysis parameters and ICSI fertilization rates. RESULTS(S): The mean (+/- SD) percentage of sperm with fragmented DNA was 14.5% +/- 1.5% and ranged from 0.5% to 75%. A significant negative association was found between the percentage of sperm with DNA fragmentation and the ICSI fertilization rate. We also observed that the motility and morphology of the ejaculated sperm were correlated negatively with the percentage of DNA fragmentation in the washed sperm recovered by the swim-up technique. CONCLUSION(S): Our results suggest that when poor-quality semen samples are used for ICSI, there is a greater likelihood that some sperm selected for injection, despite appearing normal, contain fragmented DNA. Whether sperm DNA damage may contribute to failure of pronuclear formation and embryo development in some apparently unfertilized ICSI oocytes is unclear.

The effects of trinitrotoluene toxicity on zinc and copper metabolism.

Wistar rats were exposed to trinitrotoluene (TNT, 200 mg/kg/d x 6 d/wk) for 6 weeks, and a 2-week recovery period was continued after treatment. After initiation of TNT exposure, zinc and copper concentrations in rat testes, liver and serum, and serum caeruloplasmin activity were assayed every 2 weeks. Testicular size and zinc concentrations were decreased throughout the exposure and recovery periods. After 6 weeks of TNT exposure, rat testicular copper and serum zinc concentrations were significantly decreased and increased, respectively. The activities of caeruloplasmin were decreased after 4 and 6 weeks of exposure. Liver zinc and copper concentrations and serum copper concentrations had not changed significantly in rats throughout the exposure and recovery periods. Copper concentration in hair samples from TNT-exposed workers was lower than that in control workers, but zinc content was significantly increased compared with that in control workers.

Traditional Chinese medicine (tongue acupuncture) in children with drooling problems.

Tongue acupuncture is an innovative technique in traditional Chinese medicine. We have demonstrated that specific tongue acupoints are related to various functional domains. This study aimed to assess the efficacy of tongue acupuncture in children with neurologic disability who had severe drooling problems. We conducted an intent-to-treat study in a cohort of 10 children. A continuous course of tongue acupuncture was performed daily to five acupoints in the tongue for a total of 30 sessions. Standardized outcome measures of drooling were evaluated by a blinded assessor to study the efficacy at baseline and after a course of treatment. Statistically significant improvement was noted in the following outcome measures: (1) mean visual analog scale (VAS) decreased from 6.6 (pre-TAC) to 4.67 (post-TAC) (P = 0.002); (2) mean drooling quotient (DQ) decreased from 14.016% (pre-TAC) to 8.335% (post-TAC) (P = 0.0078); and (3) mean drooling score (DS) decreased from 7.4 (pre-TAC) to 4.4 (post-TAC) (P = 0.002). This study demonstrated the efficacy of tongue acupuncture as an adjunctive or alternative treatment for patients with drooling problems and can be integrated as part of the oromotor stimulation program, drooling program, and behavioral modification program before subjecting the patient to invasive surgical procedures on the salivary glands.

Pharmacokinetics of recombinant human basic fibroblast growth factor in mice using a radioiodination method combined with SDS-PAGE and a sandwich enzyme-linked immunosorbent assay.

Pharmacokinetics of recombinant human basic fibroblast growth factor (rhbFGF) in mice were studied by using a radioiodination method combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and a sandwich enzyme-Linked immunosorbent assay (ELISA). RhbFGF concentration in serum were dertermined after iv administration of 2.5, 5 and 10 microg/kg of 125I-rhbFGF or rhbFGF. The AUCs were linearly correlated to doses. Concentrations in tissues, recoveries of 125I-rhbFGF in urine and feces were also analyzed following iv dose of 5 microg/kg. Pharmacokinetic profiles of radiolabelled and unlabelled rhbFGF were consistent in trend but exit certain differences. Mice cleared unlabeled rhbFGF markedly faster than their radiolabeled counterparts.

Identification of phase I and phase II metabolites of Guanfu base A hydrochloride in human urine.

Guanfu base A is a novel arrhythmic drug candidate isolated from the tuber of a traditional Chinese herb. Phase I and Phase II metabolites of Guanfu base A (GFA) Hydrochloride were studied in human urine by means of liquid chromatography mass spectrometry (LC/MSD) and tandem mass spectrometry (MS/MS). For phase I metabolites, Guanfu base I (GFI) was separated by HPLC and identified by comparison with authentic reference for their retention times, molecular ion peaks, fragment ions, and UV spectra. GFA oxide was also indicated to exist in human urine. For phase II metabolites, after human urine was treated either with glucuronidase or sulfatase, GFA occured in the chromatograms. It was suggested that there were GFA glucuronide and GFA sulfate in human urine. Further more, positive molecular ions, m/z 606 and m/z 510, of the two conjugates were detected in human urine by LC/MSD. In addition, characteristic ion of m/z 606 was identified as the precursor ion of m/z 177 [Glucuronic acid+H]+ by using MS/MS. Characteristic ion of m/z 430 [GFA+H]+ was also identified as a product ion of m/z 606 [GFA glucuronide+H]+. It was concluded that there were GFI. GFA oxide, GFA glucuronide and GFA sulfate in human urine.

[Involvement of the nerve fiber elements in the anterior pituitary in the regulation of ACTH secretion].

While peptidergic nerve fibers have been immunoreactively identified in the adenohypophysis of mammals, the function of these fibers in the regulation of hormone secretion remain unknown. The present work was undertaken to investigate possible modulating effects of these fibers on the secretion of ACTH in the anterior pituitary. Electrical field stimulation (EFS) was applied to excite the nerve terminals in anterior pituitary quarters of male rats under static incubation, and the ACTH level was determined by radioimmunoassay. It was found that the ACTH release from the anterior pituitary was significantly enhanced by EFS (30 mA, 0.5 ms and 10 Hz), which could be significantly decreased by adding TTX or veratridine into the incubation medium. However, TTX failed to inhibit AVP-induced ACTH secretion. Since the dispersed cultured anterior pituitary cells did not respond to the same EFS, it was suggested that there might exist dual modulatory mechanisms on the anterior pituitary hormone secretion.

Investigation of characteristics of feather follicle stem cells and their regeneration potential.

Feather follicles have the extraordinary ability to regenerate and undergo molting cycles. Being tissue-specific stem cells, feather follicle stem cells (FFSCs) have a strong capacity for proliferation and are presumed to be progenitor cells for various epidermal organs. In order to characterize FFSCs and to understand how the feather epidermis and FFSCs produce such a reliable differentiation program resulting in the formation of complex feathers, We developed a culture scheme to select and expand FFSCs from chick feather follicles. FFSCs were examined with cell profiles, mutilpotential differentiation and immunocytochemical staining. FFSCs from a single clone were capable of self-renewal and proliferation. These cells expressed integrin ß1, CD49c, cytokeratin 15 (K15), cytokeratin 19 (K19) and a neural-genic cell marker, nestin, but not a teminal differentiation-related keratinocyte marker, cytokeratin 10 (K10). FFSCs could trans-differentiate into adipocytes, neurocytes and keratinocytes. The formation of micro-feather like structures ex-vivo also revealed the potential of regeneration. These results demonstrate that FFSCs possess the properties of stem/progenitor cells and may therefore serve as a useful model for studying mechanisms of stem cell differentiation and their involvement in organ regeneration.

Detection of deoxyribonucleic acid fragmentation in human sperm: correlation with fertilization in vitro.

The objective of this study was to determine the incidence of DNA fragmentation in human sperm, and to correlate any detected DNA damage with semen analysis parameters and fertilization rates in in vitro fertilization (IVF). A total of 298 semen samples were collected from men in the infertility program at The Toronto Hospital. For each sample, the percentage of sperm with DNA fragmentation was determined using the method of terminal deoxynucleotidyl transferase-mediated dUTP-biotin end-labeling (TUNEL) and fluorescence-activated cell sorting. The percentage of sperm with fragmented DNA was less than 4% in the majority of samples but ranged from 5% to 40% in approximately 27% of the samples. A negative correlation was found between the percentage of DNA fragmentation and the motility, morphology, and concentration of the ejaculated sperm. In 143 IVF samples, a significant negative association was also found between the percentage of sperm with DNA fragmentation and fertilization rate (p = 0.008) and embryo cleavage rate (p = 0.01). In addition, 35 men who smoked demonstrated an increased percentage of sperm with fragmented DNA (4.7 +/- 1.2%) as compared to 78 nonsmokers (1.1 +/- 0.2%; p = 0.01). These results demonstrate a negative association between semen analysis parameters and sperm with fragmented DNA. Since extremely poor semen samples are the indication for intracytoplasmic sperm injection, there is a high likelihood that sperm with fragmented DNA may be selected by chance and used for oocyte injection, resulting in poor fertilization and/or cleavage rates.

Immunoreactive inhibin levels during ovarian stimulation may predict granulosa cell maturity.

OBJECTIVE: The aim of the study was to assess whether the immunoreactive inhibin response to ovarian stimulation in polycystic ovarian syndrome was of predictive value for the outcome of ovulation induction. DESIGN: Daily injections of purified FSH (Metrodin, Serono Laboratories, UK) were administered for the purpose of inducing development and ovulation of a single follicle. PATIENTS: All patients had anovulatory infertility secondary to polycystic ovarian syndrome and were resistant to clomiphene citrate. MEASUREMENTS: Alternate day serum samples were obtained for measurement of gonadotrophins, sex steroids and inhibin by radioimmunoassay. Alternate day ovarian ultrasound scans were carried out to monitor follicular development. RESULTS: There was a high incidence of multiple follicular development (MFD) (10.2 +/- 7.1 (standard deviation) follicles). There was a close relationship between the number of follicles on the day of the ovulatory trigger and serum oestradiol (E2) levels (R = 0.97, P < 0.001). This relationship was not seen with serum immunoreactive inhibin levels (R = 0.31). The cycles were divided into two groups depending whether there was or was not MFD (more than five follicles > 7 mm in diameter on day of hCG administration). The MFD group had 15.6 +/- 8.5 follicles and the non-MFD group 3.2 +/- 1.6 follicles. There was no difference in the circulating E2 concentration (pmol/l) per follicle between the two groups (non-MFD cycles, n = 6,345 +/- 28.9; MFD cycles, n = 8, 308 +/- 40.29). However, the immunoreactive inhibin concentration (U/ml) per follicle was lower, P < 0.001, in the MFD group (4.3 +/- 1.6 vs 6.9 +/- 0.5). The maximum and mean follicle diameter were lower, P < 0.01, in the MFD group (maximum follicle size 16.5 +/- 3.9 vs 21.2 +/- 1.5 mm, mean follicle size 11.5 +/- 2 vs 14.9 +/- 2.3). When individual cycles were examined E2 and immunoreactive inhibin secretion rose in parallel in cycles with less than five follicles with a rapid rise occurring when the follicles reached about 12 mm in diameter. In cycles where there was MFD there was a disparity between E2 and immunoreactive inhibin secretion with E2 levels rising 4.3 +/- 1.4 days before immunoreactive inhibin levels. CONCLUSION: These data suggest that in cycles where there are multiple small follicles, E2 secretion is maintained whereas immunoreactive inhibin secretion is substantially lower. Thus, in view of the disparity between E2 and immunoreactive inhibin secretion evident in the MFD group, measurement of immunoreactive serum inhibin concentration may be a better indicator of granulosa cell maturity. Immunoreactive inhibin secretion may occur only from healthy mature follicles.