Huperzine A is a potent,reversible,and blood-brain barrier permeable acetylcholinesterase irhibitor.The aim of this study was to compare the pharmacokinetics,tolerability,and bioavailability of two formulations with the established reference formulation of huperzine A in a fasting,healthy Chinese male population.This was a randomized,single-dose,3-period,6-sequence crossover study.The plasma concentrations of huperzine A were determined by liquid chromatography tandem mass spectrometry.Tolerability was assessed based on subject interview,vital sign monitoring,physical examination,and routine blood and urine tests.The mean (SD) pharmacokinetic parameters of the reference drug were Cmax,1.550 (0.528) ng/mL;t1/2,12.092 (1.898) h;AUC0-72h,17.550 (3.794) ng.h/mL.Those of the test formulation A and test formulation B were Cmax,1.412 (0.467),1.521 (0.608) ng/mL;t1/2,12.073 (2.068),12.271 (1.678) h;AUC0-72h,15.286 (3.434) ng.h/mL,15.673 (3.586) ng.h/mL.The 90％ confidence intervals for the AUC0-72h and Cmax were between 0.80 and 1.25.No adverse events were reported by the subjects or found with results of clinical laboratory test.The test and reference products met the regulatory criteria for bioequivalence in these fasting,healthy Chinese male volunteers.All three formulations appeared to be well tolerated.

Objective To investigate the impact of degree of hearing loss on auditory brainstem response predictions of behavioral thresholds. Methods A total of 179 patients with different hearing loss degree ( a total of 311 ears) patients from January 2012 to January 2016 in the affiliated hospital of Yan’an university were treated with auditory brainstem response (ABR), performed threshold behavior test, analyed the auditory brainstem electric response and behavioral threshold correlation, the factors that affect the ABR thresholds and behavioral threshold value and threshold of ABR and correction factor as well as linear regression equation for different frequency stimulation. Results ABR threshold was positively related to threshold behavior for the different frequencies stimulation, ABR threshold may predicte threshold behavior( P＜0.05) ; ABR threshold value can predicte behavioral threshold differ-ence error and the degree of hearing loss( P＜0.05) , through the relevant correction factors can reduce the predic-tion error. Conclusions Auditory brainstem responses may effectively predicte the hearing loss in children with be-havioural thresholds, differences in degree of hearing loss influence auditory brainstem response and behavioral threshold value, constant correction coefficient is conducive to auditory brainstem electric response and facilitates accuracy of the behavior threshold.

The present study was aimed to analyze the immunogenicity of recombinant human zona pellucida-3 peptides (r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348)) expressed in E. coli through immunizing rabbits, and to evaluate the efficacy of their polyclonal antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding respectively. Male New Zealand rabbits were immunized using r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) as antigen respectively, which was purified through an improved method of preparative gel polyacryulamide gel electrophoresis. The antibody response level of r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) immunogen in rabbits was determined by ELISA using mouse ZP3-5 (amino acid sequence(137 approximately 150) being completely conserved with huZP3(138 approximately 151) sequence) and specific huZP3-14 (amino acid sequence(327 approximately 340)) synthetic peptides as coating antigens respectively. The immunoreactivity and specificity of the anti-r-huZP3a(22 approximately 176) and anti-r-huZP3b(177 approximately 348) antisera with each r-huZP3 peptides, were tested by immunoblot and immunohistochemistry (using native huZP and human ovary section) respectively. A competitive hemizona assay (HZA) was used to evaluate the efficacy of the antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding. Both r-huZP3 peptides were able to induce higher antibody titers in rabbits. Each antiserum could specifically recognize or bind to each target r-huZP3 peptide expressed in E. coli and native human ZP in vitro. The antisera also inhibited sperm-egg binding in the HZA. These results show that r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) are of strong immunogenicity. They can be used to develop a kit for detecting whether there are autoantibodies to zona pellucida in unexplained infertile women, and their antisera might be useful tools for determining minimal B-cell epitope sequences of several known huZP3 epitope peptides.
Animals ; Egg Proteins ; biosynthesis ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Immune Sera ; immunology ; Immunization ; Male ; Membrane Glycoproteins ; biosynthesis ; genetics ; immunology ; Rabbits ; Receptors, Cell Surface ; biosynthesis ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Sperm-Ovum Interactions ; immunology ; Zona Pellucida Glycoproteins

OBJECTIVETo investigate the effects of smoking on sperm apoptosis and semen quality of healthy adult males in the main urban area of Chongqing.

METHODSAccording to the smoking habit, we divided 235 healthy adult males into a non-smoking group (n = 89) and a smoking group (n = 146). Then we detected the routine semen parameters by the computer-assisted semen analysis system and obtained the parameters of sperm apoptosis (the ratios of AN-/PI-, AN+/PI-, AN+/PI+ and AN-/PI+ sperm) by flow cytometry combined with Annexin V-FITC/PI fluorescence staining.

RESULTSThe rate of early apoptotic sperm (AN+/PI-) was higher in the smoking than in the non-smoking group ([8.1 +/- 5.1]% vs [6.8 +/- 3.8]%; P = 0.039), but there were no significant differences between the two groups in the rate of late apoptotic sperm (AN+/PI+) ([5.6 +/- 5.2]% vs [5.5 +/- 5.1]%; P = 0.87), as well as in such routine semen indexes as semen volume, sperm density, sperm motility, sperm vitality and normal sperm morphology (P = 0.30, 0.82, 0.37, 0.81 and 0.84, respectively).

CONCLUSIONThe rate of early apoptotic sperm is higher in smokers than in non-smokers, suggesting that smoking may induce early damage to sperm cells. Compared with routine semen parameters, sperm apoptosis is a more sensitive biomarker to reflect smoking-induced damage to sperm.

Apoptosis ; China ; Humans ; Male ; Semen ; Semen Analysis ; Smoking ; Sperm Count ; Sperm Motility ; Spermatozoa ; cytology

OBJECTIVETo investigate the viral etiology in hospitalized children with acute lower respiratory tract infections (ALRTI) plus platelet disorders.

METHODSA total of 255 children with ALRTI plus platelet disorders and 442 children with ALRTI and normal platelets, all of whom were hospitalized between March 2010 and February 2011, were included in the study. Their nasopharyngeal aspirate samples were collected, and RT-PCR or PCR was performed to detect 14 viruses.

RESULTSOf 255 ALRTI patients with platelet disorders, thrombocytosis was found in 253 cases (99.2%) and thrombocytopenia in 2 cases (0.8%). Among ALRTI patients with platelet disorders, 173 (67.8%) were infected with at least one virus, with human rhinovirus as the most common one, followed by parainfluenza virus type 3 (PIV3) and respiratory syncytial virus (RSV). The detection rate of PIV3 in the abnormal platelet group was significantly higher than in the normal platelet group (P<0.05). In contrast, the detection rate of influenza virus B (IFVB) in the abonormal platelet group was significantly lower than in the normal platelet group (P<0.05). The age distribution showed significant difference between the abnormal and normal platelet groups (P<0.01). Platelet disorders were mainly found in children under one year of age (P<0.01).

CONCLUSIONSThrombocytosis is often found in children with ALRTI caused by viruses, especially PIV3, but infection with IFVB seldom causes platelet disorders. Hospitalized children with ALRTI under one year tend to develop platelet disorders.

Acute Disease ; Adolescent ; Age Factors ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Respiratory Tract Infections ; blood ; complications ; virology ; Thrombocytopenia ; etiology ; Thrombocytosis ; etiology

OBJECTIVETo discuss the reliability of SARS-CoV antibody detection for SARS diagnosis.

METHODSUsing SARS-CoV ELISA kit to detect relevant antibody in fresh serum of healthy, fever, probable, and suspect cases.

RESULTSThe positive rate is 0%, 40%, and 95% respectively in healthy, probable, and suspect cases.

CONCLUSIONSIt is reliable to detect SARS-CoV antibody in late suspect patients, but there will be high false-positive result in ordinary fever cases.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Viral ; blood ; Diagnosis, Differential ; Enzyme-Linked Immunosorbent Assay ; Female ; Fever ; diagnosis ; Humans ; Male ; Middle Aged ; SARS Virus ; immunology ; isolation & purification ; Severe Acute Respiratory Syndrome ; diagnosis ; immunology ; Time Factors

Objective To summarize the treatment principle of acupuncture-moxibustion in treating Bi-impediment syndrome from the application rules of meridians and acupoints in Ming-Qing Dynasties by sorting out and analyzing the Chinese medicine literatures about acupuncture-moxibustion for Bi-impediment syndrome in Ming-Qing Dynasties, for providing literature evidence for basic and clinical research of Bi-impediment syndrome.Method Via electronic retrieval ofZhong Hua Yi Dian (Zhen Jiu Tui Na Lei) (Chinese Medical Encyclopedia,Chapter of Acupuncture-Moxibustion and Tuina), the data related to Bi-impediment syndrome in Ming-Qing Dynasties were extracted to establish a database categorized by meridians and acupoint features in Excel for analysis.Result There were 267 items of records about acupuncture-moxibustion in treating Bi-impediment syndrome in Ming-Qing Dynasties, involving the fourteen ordinary meridians, and 131 acupoints including 5 extra points; the frequency of using the Gallbladder Meridian ranked the top, followed by the Large Intestine Meridian; points from the Bladder Meridian were predominant, followed by the Gallbladder Meridian; there were 28 commonly-used acupoints (frequency>5), which were Quchi (LI 11, 26 times), Huantiao (GB 30, 23 times), Hegu (LI 4, 22 times), Chize (LU 5, 16 times),Yanglingquan (GB 34, 15 times), and Weizhong (BL 40, 14 times). Of the specific acupoint, the five Shu points were most frequently used, with a frequency of 217.Conclusion In the treatment of Bi-impediment syndrome with acupuncture-moxibustion, doctors in Ming and Qing Dynasties selected yang meridians more often than yin meridians, and Gallbladder, Large Intestine and Bladder Meridians had comparatively higher frequencies; regarding the application of acupoints, the specific acupoints were often used, especially the five Shu acupoints. The study results provide reference for acupoint selection in the treatment of Bi-impediment syndrome with acupuncture-moxibustion.

Purpose To investigate the clinical pathology significance of epithelial cellular adhesion molecule (EpCAM, CD326) expression in colorectal carcinoma cells. Methods Flow cytometry and immunofluorescence assay were used to detect EpCAM expression in 55 cases of fresh colorectal cancer tissues and adjacent normal mucosa. The percentage of positive cells (PPC) and mean fluorescence index (MFI) were calculated. Correlation of EpCAM expression with DNA ploidy change and its value were investigated in the early diagnosis of colorectal cancer. Results The values of PPC and MFI were significantly higher in colorectal carcinoma tissue than that in the normal colorectal tissue [(PPC: (83.48 ± 7.07)％ vs (43.56±5.29)％, t =39.22, P＜0.001. MFI: 28.90(19.60-45.89) vs4.89(3.79-6.28), Z=-6.45, P＜0.001) ]. There were also significant differences (P＜0.01) in the values of PPC and MFI between invasive type and ulcer types, between well-and moderately-differentiated and poorly-differentiated cancers, between Dukes stages A + B and C + D, (between pTNM stages I+ II and III + IV, between pTl + T2 and pT3 + T4, and between pNO and pNl. DNA content analysis showed that DNA polyploid was detected in 39 of 55 colorectal carcinoma (70.90％ ). The DNA index (DI) and ploid were correlated with differentiation degree and Dukes stages, but uncorrelated with lymph node metastasis. At the same time, in the EpCAM positive cases, DI was increased with increased expression of EpCAM (r = 0.668, P =0.000) and the proliferation index of cells in S phase ratio(Sphase fraction, SPF) (r1 =0.664, P1 =0.000, r2 =0.651, P2 = 0.000 ). Conclusion EpCAM expression is obviously upregulated in colorectal carcinoma, and it is closely correlated with the invasion, metastasis and proliferation of tumor cells. The combination of EpCAM expression and DNA content analysis provides references to the early diagnosis and prognosis evaluation in colorectal carcinoma.

Objective To detect the uncontrolled new psychoactive tryptamines involved in drug-related cases with high resolution mass spectrometry and nuclear magnetic resonance spectroscopy. Methods White and brown powder obtained in actual cases were extracted and analyzed by gas chromatography-quadrupole time-of-flight mass spectrometry （GC-QTOF-MS）, ultra-high performance liquid chromatography-linear ion trap quadrupole-orbitrap mass spectrometry （UPLC-LTQ-Orbitrap MS） and ¹H-nuclear magnetic resonance spectroscopy （1H-NMR）. Results After detection by GC-QTOF-MS, the components of white powder showed main characteristic fragment ion peaks at m/z 218.141 0 （molecular ion peak）, 72.080 6 （base peak）, etc. After detection by UPLC-LTQ-Orbitrap MS, its protonated molecular ion was m/z 219.149 4. The main ions in the secondary mass spectrum under the collision-induced dissociation （CID） mode were m/z 160.076 3 and 72.080 8. After detection by GC-QTOF-MS, the components of brown powder showed main characteristic fragment ion peaks at m/z 246.135 7 （molecular ion peak）, 58.065 1 （base peak）, etc. After detection by UPLC-LTQ-Orbitrap MS, its protonated molecular ion was m/z 247.145 0. The main ions in the secondary mass spectrum under CID mode were m/z 202.087 1, 160.076 3 and 134.060 5. NIST 17 library retrieval and ¹H-NMR confirmed that the white powder and brown powder contained new psychoactive tryptamines 4-OH-MET and 4-AcO-DMT, respectively. Conclusion GC-QTOF-MS, UPLC-LTQ-Orbitrap MS and ¹H-NMR can be used together to identify unknown new psychoactive substances.
Chromatography, High Pressure Liquid ; Gas Chromatography-Mass Spectrometry ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Tryptamines

To screen the anti-inflammatory active fraction of unripe Forsythiae Fructus, and elucidate the action mechanism, water decoction, ethyl acetate portion, n-butanol portion and residue water extracts of unripe Forsythiae Fructus were administered into rats for continuously 15 days. The acute lung injury inflammatory model was established to observe the section structure of lung tissues. Levels of IL-6, TNF-α, IL-1β and IL-10 in bronchoalveolar lavage fluid were determined by ELISA kits, and changes in endogenous metabolites in serum were analyzed based on 1H-NMR metabolomics. The results showed that ethyl acetate portion of unripe Forsythiae Fructus had a better anti-inflammatory activity against acute lung injury, and could suppress the release of inflammatory factors of IL-6, TNF-α, IL-1β, significantly reduce contents of creatine, β-OH-butyrate, succinate, lysine, valine, isoleucine and glutamine, and elevate the content of GPC in serum. Ethyl acetate portion was proved to be the main fraction of anti-inflammatory activity from the perspective of endogenous metabolites in serum, and played an anti-inflammatory role by regulating creatine metabolism, choline metabolism, branched-chain amino acid metabolism and TCA cycles. This study could lay a foundation for studying pharmacodynamic material basis of unripe Forsythiae Fructus.