This study is to establish the methods for determination of iridoid glycosides and triterpenic acids in Corni Fructus and provide technical support for the quality control of Corni Fructus. Morroniside, loganin and sweroside were determined by HPLC-UV method with acetonitrile and 0.1% formic acid as the mobile phase, and the detective wavelength was set at 240 nm. Oleanolic acid and ursolic acid were determined by HPLC-ELSD method with methanol-0.5% ammonium acetate (87:13) as the mobile phase. The results showed that the linear ranges of morroniside, loganin and sweroside were 5.335-213.4 mg · L(-1) (r = 0.9999), 5.515-220.6 mg · L(-1) (r = 1.0000), 1.992-79.68 mg · L(-1) (r = 1.0000), respectively. The average recoveries of the above three iridoid glycosides were 98.49%-99.28% with RSDs of recoveries being less than 2%. The linear ranges of oleanolic acid and ursolic acid were 7.74-154.8 mg · L(-1) (r = 0.9964), 10.82-216.4 mg · L(-1) (r = 0.9996), respectively. The average recoveries of the above two triterpenic acids were 98.11%-99.27% with RSDs of recoveries being less than 3%. The method established in this research is simple, rapid and reliable, and can be used for quality control of Corni Fructus. Furthermore, the research provided experimental data for the improvement of present quality standard of Corni Fructus, which has important significance to guarantee its quality and clinical curative effect.
Cornus ; chemistry ; Drug Stability ; Oleanolic Acid ; analysis ; Quality Control ; Triterpenes ; analysis

To establish the online two-dimensional liquid chromatography by using double gradient liquid chromatography system and UV detector, in order to simultaneously determine the content of paeoniflorin, paenol, amygdaloside and cinnamic acid. A pump of the two-dimensional liquid chromatography was adopted as the one-dimensional separation pump. C18 (3.0 mm x 150 mm, 3 microm) was used as the analytical column, with acetonitrile as the organic phase and 0.08% phosphoric acid + 0.08% triethylamine as the aqueous phase for gradient elution at the flow rate of 0.5 mL x min(-1). Another pump of the two-dimensional liquid chromatography was adopted as the two-dimensional separation pump. PAII C18 was used as the analytical column, with acetonitrile as the organic phase and 20 mmol, pH 3.0 monopotassium phosphate as the aqueous phase for gradient elution at the flow rate of 0.8 mL x min(-1). The detection wavelengths were set at 218, 230, 275 nm by using wavelength time-switching program. The linearity range of paeoniflorin, amygdaloside, paeonol and cinnamic acid were 5.55-222 (r = 0.999 7), 6.6-264 (r = 0.999 8), 3.3-132 (r = 0.999 5) and 0.315-12.6 mg x L(-1) (r = 0.999 7), respectively. The average recoveries of the four components were between 96.12% and 103.9%. The experiment proved that this method was so rapid and accurate in determination results that it could be used for evaluating drug quality.
Capsules ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Online Systems ; Time Factors

Objective To investigate the ionic mechanism of remifentanil-induced vaso-relaxation by recording calcium-activated K~+-currents(IKCa)in human arterial smooth muscle cells(MASMCs).Methods Human MASMCs were obtained freshly by the method of enzymolysis.IKCa were separated with 4-AP 5 mmol?L~(-1) and recorded by the whole-cell voltage-clamped technique.The effects of remifentanil(1.2,4.8,19.4 mmol?L~(-1))on amplitude and threshold of activation for IKCa were assessed.Results Remifentanil significantly increased IKCa and had no effect on the threshold of activation for IKCa as compared with the control group(P＜0.05 or 0.01),and the IKCa returned to the baseline level after remifentanil was washed out.Conclusion Remifentanil produces vasodilation by activating IKCa in MASMCs.

The gene complex of Human Leukocyte Antigen (HLA) is located of chromosome 6p21, which is the most complicated dominant polymorphic genetic system. The HLA system has 108 genotypes. It is the best human genetic marker. It has been applied to forensic paternity test and individual identification. This article discusses the research development of HLA polymorphism and its application in forensic medicine.
Forensic Medicine ; Gene Frequency ; HLA Antigens/genetics* ; HLA-A3 Antigen/genetics* ; Humans ; Polymorphism, Genetic

Objective To investigate the clinical diagnosis,treatment and prevention of fallopian tube prolapse(FTP)after hysterectomy.Methods A total of 7949 patients received hysterectomy from 1983 to Aug 2005 in Peking Union Medical College Hospital,including 6229 cases of trans-abdominal hysterectomy(TAH),780 cases of transvaginal hysterectomy(TVH),and 940 cases of laparoscopic assisted vaginal hysterectomy(LAVH).Nine cases(including 1 case from other hospital)of FTP after hysterectomy were analyzed retrospectively for their symptoms,diagnosis and treatment.All of them were diagnosed according to the results of histology and follow-up.Results The overall incidence of FTP after hysterectomy was 0.11%(9/7949).Incidence of FTP after trans-abdominal hysterectomy was 0.08% (5/6229),after vaginal hysterectomy 0.51%(4/780),and after laparoscopic assisted vaginal hysterectomy 0(0/940).There were no symptoms in 3 cases,but the other 6 cases had symptoms.The pelvic examination revealed the typical prolapsed fimbrial end of a fallopian tube in 3 cases and red granulation tissue in the other 6 cases.All of them were excised vaginally and cauterized.The results were confirmed by histological examination.No recurrent cases were reported in follow up.Conclusions FTP is a rare complication after hysterectomy.The prognosis is well after proper diagnosis and treatment.Salpingectomy or fixation of accessories into the pelvic wall and complete peritonealisation at the time of hysterectomy are important methods to prevent FTP after hysterectomy.

Seven acylated triterpene saponins were isolated from the roots of Securidaca inappendiculata by means of various chromatographic techniques such as silica gel, MPLC, preparative HPLC, and semi-preparative HPLC. Their chemical structures were identified as securioside A(1), securioside B(2), 3-O-β-D-glucopyranosyl presenegenin 28-O-β-D-xylopyranosyl-(1-->4)-α-L-rhamnopyranosyl-(1-->2)-[β-D-glucopyranosyl-(1-->3)]-4-O-[(E)-3,4-dimethoxycinnamoyl]-β-D-fucopyranosyl ester(3), 3-O-β-D-glucopyranosyl presenegenin 28-O-β-D-xylopyranosyl-(1-->4)-α-L-rhamnopyranosyl-(1-->2)-[β-D-glucopyranosyl-(1-->3) ] -4-O-[(E/Z)-3, 4-dimethoxycinnamoyl]-β-D-fucopyranosyl ester(3/4), 3-O-β-D-glucopyranosyl presenegenin 28-O-α-L-arabinopyranosyl-(1-->3)-β-D-xylopyranosyl-(1-->4)-α-L-rhamnopyranosyl-(1-->2)-4-O-[(E)-3,4-dimethoxycinnamoyl]-β-D-fucopyranosyl ester(5), polygalasa- ponin XLV(6), and polygalasaponin XLVI (7) on the basis of spectroscopic data analysis and physicochemical properties. Among them, compounds 5-7 were isolated from the plants in genus Securidaca for the first time and compounds 3, 3/4 were isolated from the species for the first time. The cytotoxicity assay showed that compounds 2, 3/4, 5 have moderate cytotoxic activities against Lewis lung carcinoma LLC cells with IC50 values of 41.10, 38.17, and 48.92 µmol · L(-1), respectively; compound 2 exhibited moderate cytotoxic activities against human breast cancer MCF-7 cells with an IC50 value of 47.93 µmol · L(-1).
Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Humans ; MCF-7 Cells ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification ; pharmacology ; Securidaca ; chemistry

OBJECTIVETo observe the effect of electro-acupuncture (EA) on alteration of immune function of patients undergoing open-heart surgery with cardiopulmonary bypass (CPB), and to appraise the value of acupuncture-drug compound anesthesia in the operation.

METHODSThirty patients undergoing atrial septal defect repairing operation were selected and divided into three groups, Group A was the general anesthesia group; Group B, the acupuncture anesthesia group and Group C, the general anesthesia plus EA group. Peripheral venous blood of patients was collected at different time points, i.e. before anesthesia, before CPB, 30 min and 24 hrs after CPB, to determine natural killer cells activity (NKCA), and the levels of interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) in supernatant of cell culture were also tested.

RESULTSNKCA was significantly lowered in Group A before CPB but increased in Group B, while no evident change was found in Group C, so the level of NKCA in Group B was significantly higher than in the other two groups. It lowered in all the three groups after CPB, especially evidently in Group B, so as to cause the NKCA level in Group B lower than that in Group A. The lowering further progressed, 24 hrs after CPB, NKCA in Group B was more reduced than that in Group C. Levels of IFN-gamma and IL-2 lowered in all the three groups after CPB, and further lowered at time point of 24 hrs after CPB, but the parameters in Group C were significantly higher than those in Group B.

CONCLUSIONEA could enhance NKCA, but acupuncture anesthesia couldn't inhibit the suppressive effect of CPB on NKCA, IL-2 and IFN-gamma, suggesting that the immunosuppression induced by stress has a prior effect. General anesthesia plus EA yielded better effect than general anesthesia and acupuncture anesthesia, but it could't improve the immunosuppression completely, indicating that the compound anesthesia could partially improve the immunosuppression induced by CPB.

Acupuncture Analgesia ; Adult ; Anesthesia, General ; Cardiopulmonary Bypass ; Electroacupuncture ; Female ; Heart Septal Defects, Atrial ; immunology ; surgery ; Humans ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Killer Cells, Natural ; immunology ; Male ; Middle Aged

This study was aimed to compare the proportion of endothelial cells (EC) in human bone marrow mesenchymal stem cell (BM-MSC) and human umbilical cord mesenchymal stem cells (UC-MSC), and to investigate the influence of vascular endothelial growth factor (VEGF) on proportion of EC in MSC. Flow cytometry was used to detect the proportion of CD34(+)CD133(+) and vWF(+)CD31(+) double positive cells in MSC. Wright's staining was employed to observe the influence of VEGF on morphology of MSC. The expressions of CD34, CD133, CD31, vWF were detected by immunofluorescence. qRT-PCR was performed to detect the influence of VEGF on EC marker genes' expression of MSC. The results showed that there were a small amount of EC and endothelial progenitor cells (EPC) in obtained BM-MSC and UC-MSC. After exposed to VEGF 10 ng/ml for 24 h, aspect ratio of MSC and the proportion of EC increased, while proportion of EPC decreased. Expression of EC related marker genes such as Tie-2 and ecNOS up-regulated, especially in UC-MSC. It is concluded that small amount of EC and EPC exists in cultured BM-MSC and UC-MSC, VEGF can enhance the proportion and function of EC in MSC.
Antigens, CD ; metabolism ; Bone Marrow Cells ; cytology ; Cell Separation ; Cells, Cultured ; Endothelial Cells ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Umbilical Cord ; cytology ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo study the effects of VAC on starting the process of wound healing and decreasing apoptosis.

METHODSTo examine the variations in expression of proto-oncogenes c-myc, c-jun and Bcl-2 in pig wound model with acute full-thickness skin defect and human chronic wounds by immunohistochemistry, calculate the numbers of expressive positive cells and the labelling index (LI), and observe the process of wound healing.

RESULTS(1) In pig experiment, the wound in experimental group was very clean and without obvious exudates, many neoepiderm and granulation tissue rapidly appeared or formed after 6 days, and healed completely by the 25th day. On the contrary, in the wound of control group, more exudates and blood crust could be seen and fewer neoepiderm and granulation tissue appeared after 6 days and was healed by 30th day. Immediately after the wound was created, the expression of c-myc, c-jun and Bcl-2 was lower and mainly situated in nucleus or cytoplasma of the basilar cells. After the wound was created in control group, or after starting the VAC treatment in experimental group, their expression rapidly and obviously increased, the distribution of the positive cells also became enlarged, but the amount of expression decreased rapidly after the expressive peak have reached. In the successive 12 days following the wound was created, the expression of c-myc, c-jun and Bcl-2 in the experimental group was constantly higher than that of the control group. (2) In human chronic wounds, there wasn't obvious secretions and more healthy granulation tissue was rapidly formed after VAC treatment. The expression of c-jun was mainly located in cytoplasma of basilar cells of epithelium, dermal fibroblasts and inflammatory cells, and the positive cell and labelling index obviously decreased. The expression of c-myc and Bcl-2 was mainly in cytoplasma of basilar cells, but the amount of expression and the labelling index became obviously increased after VAC treatment.

CONCLUSIONSVAC could rapidly start the healing course of the pig' s acute skin wound and human chronic wound, decrease apoptosis of the reparative cells, so as to accelerate wound healing.

Adult ; Animals ; Apoptosis ; Female ; Humans ; Male ; Middle Aged ; Negative-Pressure Wound Therapy ; Proto-Oncogene Proteins c-jun ; metabolism ; Proto-Oncogenes ; Swine ; Wound Healing

OBJECTIVETo investigate the use of anterior cervical discectomy and fusion with self-locking cages to treat multi-segmental cervical myelopathy.

METHODSFrom April 2008 to March 2010, anterior cervical discectomy and fusion with self-locking cages were performed on 45 patients who suffered from multi-segmental cervical myelopathy, among of them there were 23 male and 22 female, aged from 32 to 67 years (average 53 years). Recording the Japanese Orthopedic Association (JOA) scores and SF-36 scores in the protocol time point, in order to investigate the clinical outcome, meanwhile, accumulating the pre-operation and postoperation X-ray films of cervical spine for measuring the height of intervertebral space, whole curvature of cervical spine and the rate of fusion by repeated measures analysis of variance.

RESULTSThe mean follow-up time was 28.4 months (24 - 35 months). JOA scores ascended from preoperative 6.5 ± 3.1 to postoperative 13.4 ± 1.7 (F = 17.84, P = 0.001), the 7 scores of SF-36 improved significantly after operation (t = 1.151 - 12.207, P < 0.05), but mental health not. The fineness rate was 91.1%. Height of disc space ascended from preoperative (5.5 ± 1.8) mm to postoperative (8.3 ± 0.8) mm (F = 11.71, P = 0.043), globle curvature of cervical spine ascended from preoperative 5° ± 7° to postoperative 10° ± 14° (F = 234.53, P = 0.000), the change of the two index was significantly, respectively. Fat necrosis in one case and hematoma in another case at the bone donor-site were found, both of the two cases were cured by physiotherapy. All of the 45 cases (111 segments) achieved bone fusion.

CONCLUSIONThe use of anterior cervical discectomy and fusion with self-locking cages to treat multi-segmental cervical myelopathy possess many advantages as follows: satisfactory clinical outcome, minimally invasive, higher fusion rate, higher orthopaedic ability.

Adult ; Aged ; Cervical Vertebrae ; surgery ; Decompression, Surgical ; methods ; Diskectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Internal Fixators ; Male ; Middle Aged ; Spinal Cord Diseases ; surgery ; Spinal Fusion ; instrumentation ; methods ; Treatment Outcome