OBJECTIVE Cistanche deserticola Y. C. Ma is an official plant that grows in arid or semi-arid areas.Our early work demonstrated that Cistanche extracts protect against sperm damage in mice under bisphenol A induced reproductive damage. Echinacoside (ECH)is one of the major active components. The present study investigated the mechanisms behind the possible protective effects of ECH against oligoasthenospermia in rat and identified the interaction between ECH and AR. METH-ODS The distribution of ECH was assayed by HPLC,the quantity and quality of sperm was evaluated and hormone levels were determined by radioimmunosorbent assay. The levels of androgen receptor (AR)and key steroidogenic-related genes were reduced as determined by Western blotting and qPCR analysis.The interaction between ECH and AR were evaluated by fluorescence localization assay,indi-rect ELISA and molecular docking. RESULTS ECH significantly increased the quantity of sperm and secretions of luteinizing hormone and testosterone.ECH was distributed to the hypothalamus but not in the testes.ECH combined with hypothalamic AR in the pocket of Met-894 and Val-713 to inhibit transfer of AR from the cytoplasm to nuclei in the hypothalamus.While negative feedback of sex hormone regula-tion was inhibited, positive feedback was stimulated to increase the secretion of luteinizing hormone and testosterone subsequently enhancing the quantity of sperm. C. militaris significantly alleviated the BPA-induced reproductive damage by increasing testicular superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and glutathione (GSH); as well as by reducing serum malondialdehyde (MDA). C. militaris not only obviously enhanced the levels of serum LH and T, but it also improved the sperm count and motility compared to the BPA-treated group.CONCLUSION C.militaris and ECH protect the BPA induced reproductive damage.ECH blocks AR activity in the hypothalamus to increase the quantity of sperm and protect against oligoasthenospermia in rats.

Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.

Objective To explore the impact of early repetitive painful procedures on subsequent pain behaviors and physiological indicators in full-term infants.Methods Sixty-two full-term neonates were enrolled (male 36 cases,female 26 cases) from the medical center of neonatal care at Nanjing Children's Hospital Affiliated to Nanjing Medical University from March to May in 2009.The data of all painful procedures were performed on those neonates and their responses to them were collected by using digital video recording were collected.The Neonatal Facial Coding System C(NFCS) and Neonatal Infant Pain Scale(NIPS) were used prospectively to evaluate the pain response to painful procedures.Results The average experience of pain caused by operation on newborn at the hospital was 56.5 times (12-249 times),and the daily average was 5.9 times (4-26 times).After they were exposed to more than 10 repetitive pain procedures,the full-term neonates showed the declining pain response assessed by NFCS and NIPS during the subsequent painful procedures (NFCS:P10=0.012,P20=0.015,P30=0.041;NIPS:P10=0.006,P20=0.015,P30=0.049),and the temporarily enhanced pain response was observed in the preparation phase of the subsequent painful procedures (NFCS:P2＜0.001,P3＜0.001,P4=0.004,P5=0.009;NIPS:P2＜0.001,P3＜0.001,P4=0.045,P5 =0.031).Bnt there was no difference in preparation phase.There was no alternation in latency and crying time after repetitiing painful procedures,but the proportion of crying neonates in the preparation phase of the painful procedures was increased(P=0.032).Conclusions After exposed to repetitive pain,full-term neonates expressed hypoalgesia during subsequent painful procedures,but the temporary irritation was enhanced during the following phases exposed to stress.

Professor DU Xiao-shan 's academic thought and clinical experiences was introduced in this paper. He has noble medical ethics and is fair to the patients without distinction. He has great learning and rich practical experiences, and he specializes in all kinds of common diseases as well as many difficult and complicated cases. He never stops studying the ancients to make innovation and he creates the technique of the quick-puncture and slow-twist and DU's heat-reinforce method. He teaches to carry forward acupuncture and moxibustion and his achievements have become an important part of acupuncture and moxibustion science.
Acupuncture ; education ; history ; Acupuncture Therapy ; history ; methods ; China ; History, 20th Century ; History, 21st Century ; Humans

OBJECTIVETo observe the dynamic process of basement membrane remodeling after the combined grafting of xenogenic acellular dermal matrix with autoskin.

METHODSThe rat skin wounds were covered with xenogenic porcine acellular dermal matrix overlaid with razor thin autoskin. The skin samples were collected at 1, 2, 3, 4, 8, 12 and 16 post-grafting weeks. The changes in laminin expression in the basement membrane and the ultrastructure of the basement membrane at 12 post-grafting weeks were observed by immunohistochemistry and transmission electron microscopy. The results were compared with those in simple thin autoskin grafting as the control.

RESULTSThe laminin expression in the combined grafting was higher than that in control. At 12 post-grafting weeks, the basement membrane in combined grafting rats was clear and continuous and the hemidesmosome was relatively more in amount and distributed evenly. While in the autoskin group, the lamina densa in the basement membrane was blurred and discontinuous with a decrease in and uneven distribution of hemidesmosome.

CONCLUSIONThe increased expression of laminin in the basement membrane in the combined grafting rats might be beneficial to the remodeling of the basement membrane and to strengthening the connection of epithelium to the dermis, thus wound healing quality would be improved.

Animals ; Basement Membrane ; metabolism ; ultrastructure ; Burns ; surgery ; Dermis ; transplantation ; Immunohistochemistry ; Laminin ; biosynthesis ; Male ; Microscopy, Electron ; Rats ; Rats, Wistar ; Skin Transplantation ; methods ; Swine ; Time Factors ; Transplantation, Autologous ; Transplantation, Heterologous ; Wound Healing

OBJECTIVETo investigate the dynamic process of the inflammatory response and the profile of Th1/Th2 cytokines after xenogenic acellular dermal matrix (ADM) transplantation with thin split-thickness skin autograft overlay.

METHODSSD rats were used in the study. In the control group, thin split-thickness skin autograft (STSG) was transplanted in the full-thickness skin defect of the SD rats; in the experimental group, the xenogenic acellular dermal matrix combined with thin split-thickness skin autograft was transplanted. The inflammatory response was examined histologically and Th1/Th2 cytokine mRNA expression in skin grafts was determined by reverse transcription-polymerase chain reaction.

RESULTSInflammatory reaction was induced by ADM at the early stage of transplantation and decreased gradually. Th2 cytokine mRNA expression was higher in the ADM group than that of the control group whereas the Th1 cytokine mRNA expression was undetected in both groups.

CONCLUSIONXenogenic acellular dermal matrix is immunogenic. The increased expression of Th2 cytokines may be related to the humoral immune responses and the absence of ADM graft rejection.

Animals ; Cytokines ; genetics ; Dermis ; immunology ; transplantation ; Gene Expression ; Inflammation ; immunology ; Interferon-gamma ; genetics ; Interleukin-2 ; genetics ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Skin Transplantation ; methods ; Swine ; Transplantation, Autologous ; Transplantation, Heterologous

OBJECTIVETo investigate if WAVE1 is involved in mult drug-resistance (MDR) of human leukemia cell line K562/A02.

METHODSThe level of WAVE1 in K562 and K562/A02 cells was assayed by Western blot and RT-PCR; K562 cells and K562/A02 cells were transient transfected with pEFBOS-WAVE1 reconstructed plasmid or specifically siRNA to WAVE1. 50% inhibition concentration (IC50) of doxorubicin on K562/A02 was determined by WST-8 assay. Hoechst33258 staining was used to examine cell morphological changes and to calculate percentage of apoptotic nuclei. The mRNA level of mdrl was assayed by RT-PCR. The Bcl-2 protein was assayed by Western blot.

RESULTS1. The WAVE1 expression at mRNA and protein level in K562/A02 cells was increased by about 70% and 63% respectively as compared with that in K562 cells. 2. Overexpression of WAVE1 in K562 cells by transient transfection significantly increased the resistance to doxorubicin, and increased IC50 from (0.05 +/- 0.00) microg/ml to (2.99 +/- 0.12) microg/ml, and at 1 microg/ml or 5 microg/ml of doxorubicin treatment, cell apoptotic nuclei rate was decreased by 30% or 35% respectively. 3. Suppression of WAVE1 in K562/A02 cells by siRNA resulted in a reversal of MDR to doxorubicin, and decreased IC50 from (4.29 +/- 0.15) microg/ml to (1.85 +/- 0.07) microg/ml, and at 1 microg/ml or 5 microg/ml of doxorubicin treatment, cell apoptotic nuclei rate was increased by 24% or 21% respectively. 4. Overexpression of WAVE1 in K562 cells significantly increased the mdrl mRNA and the Bcl-2 protein, and suppression of WAVE1 in K562/A02 cells by siRNA decreased the mRNA and the protein.

CONCLUSIONWAVE1 involves in the MDR mechanisms in K562/A02 leukemia cells through regulation the level of mdrl and Bcl-2.

Apoptosis ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Genetic Vectors ; Humans ; K562 Cells ; Plasmids ; genetics ; RNA Interference ; RNA, Messenger ; genetics ; Transfection ; Wiskott-Aldrich Syndrome Protein Family ; genetics ; metabolism

Objective To investigate the treatment status of antiparkinsonism in Xi'an. Methods Six general hospitals were randomly chosen in Xi' an and all Parkinson' s disease (PD) patients were interviewed by questionnaire from Jan. 2007 to Apr. 2007. Results 92 PD outpatients were enrolled in, including 48 males and 44 females, from 43 to 86 years old (mean 65.6±17.1) with duration of the disease from 0.2 to 27.8 years (mean 4.4 ±9.4). The preference of the drug use from the patients were: 40 (43.5%) preferred taking levodopa, 25 (27.2%) with amantadine and/or trihexyphenidyl, 14(15.2%) with levodopa and others, 4(4.4%) with dopamine agonist and others, 2 (2.2%) with other drugs, 7 (7.6%) with no treatment. There were 69 (75.0%) patients onset with resting tremor, 15 (16.3%) with bradykinesia, 6 (6.5%) with rigidity, and 2 (2.2%) with unknown symptoms. There was no startically significant difference in anti-PD drugs among the patients onset with different symptoms (P>0.05). 45 patients appeared the onset of disease before 65 years old and with no dementia, 47 onset after 65 with or without dementia. There was no significant difference of anti-PD drugs between the two groups (P>0.05). Most patients initiated anti-PD treatment with levodopa but few of them chose dopamine agonist. According to the classification of Hoehn & Yahr, 25(27.2%) belonged to Grade Ⅰ, 53(57.6%) to Grade Ⅱ ,8(8.7%) to Grade Ⅲ ,3(3.3%) to Grade Ⅳ and 3 (3.3%) to Grade Ⅴ. There was no significant differences of anti-PD drugs between different grades of the disease (P>0.05). 55.3% of the patients changed their anti-PD drugs randomly during the therapy, but with no relation to their gender, age, educational level, dementia, the number of family members, course of diseases, or the degree of Hoehn & Yahr, frequency and categories of medicine. Conclusion Anti-PD treatment in Xi' an did not strictly follow the standardized protocol, with few patients using dopamine agonist and over 50% of the patients changed their drugs randomly.

AIM To study the chemical constituents of the secondary metabolites of endophytic fungi DL02 from Saposhnikovia divaricata (Turcz.) Schischk.and their biological activities.METHODS The ethyl acetate and methanol extract leavening from S.divaricata was isolated and purified by silica and Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The antifungal and antibacterial activities were determined by the minimum inhibitory concentration method.RESULTS Seven compounds were isolated and identified as β-sitosterol (1),kojic acid (2),stigmasta-7,22-diene-3β,5α,6α-triol (3),uracil (4),allantoin (5),erythritol (6) and adenosine (7).Compound 4 had strong inhibitory effects on pseudomonas aeruginosa with the MIC values of 31.3 μg/mL.CONCLUSION Compounds 2-5 and 7 are isolated from this fungus for the first time.Compounds 4-5 have strong biological activities.