Content determination of four kinds of alkaloids in Uncaria by HPLC-MS-AIS / 四川精神卫生

ObjectiveTo determine the contents of four kinds of indole alkaloids （rhynchophylline， isorhynchophylline， corynoxeine， isocorynoxeine） in Uncaria by high-performance liquid chromatography-mass spectrometry-automatic internal standard （HPLC-MS-AIS）. MethodsChromatographic separation was performed using C18 column （3.0 mm×50 mm， 3.3 μm）， and the mobile phase， comprising 0.1% formic acid aqueous solution-acetonitrile （82∶18， V/V）， was eluted at a flow rate of 0.5 mL/min and column temperature of 30℃. Mass spectrometric detection was performed using an electrospray ionization source and positive multiple-reaction monitoring mode at a voltage capillary of 4 000 V. The mass-to-charge ratio （m/z） transition was 385.25/160.10 for rhynchophylline， 385.30/160.10 for isorhynchophylline， 383.25/160.15 for corynoxeine and 383.25/160.15 for isocorynoxeine， respectively. The injection volume was kept constant at 2 μL. ResultsThe linear concentration ranges of rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine were 2.30~600.00 ng/mL （r=0.999 3）， 2.30~600.00 ng/mL （r=0.999 2）， 2.47~650.00 ng/mL （r=0.999 4） and 2.47~650.00 ng/mL （r=0.999 2）， respectively. The relative standard deviation （RSD） of precision and stability were all lower than 5.00%， the accuracy ranged from 92.40% to 104.10%， and the average recovery was 95.90%~104.60%. ConclusionHPLC-MS-AIS method is simple and accurate for the determination of four kinds of alkaloids in Uncaria， and can be used as a new method for quality control of Uncaria.

Development of a 2D-HPLC for simultaneous determination of rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine / 四川精神卫生

ObjectiveTo develop a two-dimensional high-performance liquid chromatography （2D-HPLC） for simultaneous determination of the contents of four kinds of Uncaria alkaloids： rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine. MethodsThe 2D-HPLC apparatus was comprised of a first chromatographic column in version Aston SC2 （3.5 mm×25 mm， 5 μm）， an intermediate column in version Aston SH C18 （3.5 mm×10 mm， 5 μm）， and an analytical column in version Aston SCB （4.6 mm×125 mm， 5 μm）. The mobile phase of the first and second liquid chromatography system were CAA-1 and mixed mobile phase （V BPI-1 basic mobile phase ∶ V MPI-1 mobile phase ∶ V OPI-1 organic mobile phase = 45∶14∶41）. The chromatographic parameters included a flow rate of 1.0 mL/min， a column temperature of 40℃， a wavelength of 254 nm， an injection volume of 500 μL and a detection time of 9.5 min. ResultsThe linear ranges of rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine were 9.77~10 000.00 ng/mL （r=0.999 6）， 10.74~11 000.00 ng/mL （r=0.999 7）， 10.74~11 000.00 ng/mL （r=0.999 7）， 10.74~11 000.00 ng/mL（r=0.999 6）， respectively. The relative standard deviation （RSD） of precision， stability and repeatability were all less than 5.00%. The accuracy was 95.20%~104.01%， and the recovery rate was 93.63%~101.38%. ConclusionThe 2D-HPLC developed for simultaneous determination of four kinds of alkaloids in Uncaria is simple and accurate， which can be used as a new method for quality control of Uncaria.

Academic thoughts and related research progress of Guarding field theory in traditional Chinese medical surgery / 国际中医中药杂志

The Guarding field theory is a unique theoretical system of traditional Chinese medicine for the treatment of surgical diseases such as scabies, sores and gangrene. The hooping in place is the best performance of Guarding field theory. This paperreviews the research progress of Guarding field theory from the aspects of theoretical research, basic research, clinical research in recent years, which shows the connotation of this theory has been expanded. and the generalized concept of Guarding field theory refers to human's healthy Qi, and the field theory has been widely used in treating diseases likemalignant tumor, postoperative anorectal infection, refractory wound to guide clinical treatment.

Metformin improving anxiety-like behaviors through regulation of autophagy following focal cerebral ischemia in rats / 中国生化药物杂志

Objective To investigate probable effect of metformin on anxiety and autophagy pathway in rats with focal cerebral ischemia . Methods The rats were divided into seven groups:shame group, ischemia group, metformin group, compound C group, compound C +ischemia group, metformin+ischemia group, and metformin+compound C+ischemia group.The rats were administered with metformin 14 days and compound C 30 min pretreatment.The blood glucose level, body mass, neurological score and indicators of elevate plus maze test were recorded of two groups after 72h ischemia-perfusion.The autophagy-related protein of Atg-7, Atg-5, beclin-1 were analysed by Western blot.ResuIts Metformin had no significant effect of body mass and blood glucose level.The prior administration of metformin significantly elevated neurological score in ischemia group (P<0.01).The elevate plus maze test results showed that prior administration of metformin significantly elevated percentage of open-arms detention time in ischemia group (P<0.01), but there was no significant difference in open-arms times.Western blot result showed that prior administration of metformin significantly elevated Atg-7, Atg-5, beclin-1 protein expression in ischemia group (P<0.001).Compound C resist the above effects of metformin (P<0.01).ConcIusion The metformin improves psychological disorders and movement disability through regulation of autophagy following focal cerebral ischemia in rats.

Comparison of neonatal tolerance to thoracoscopic and open repair of esophageal atresia with tracheoesophageal fistula / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Advances in minimally invasive surgical techniques and neonatal intensive care for neonates have allowed for repair of the neonatal esophageal atresia with tracheoesophageal fistula (EA/TEF) to be approached endoscopically. However, thoracoscopic surgery in children is still performed in only a few centers throughout the world. The aim of this study was to compare the neonatal tolerance to the thoracoscopic repair (TR) and the open repair (OR) and also to discuss anesthetic management in thoracoscopic procedure.</p><p><b>METHODS</b>We performed a prospective study enrolling newborns diagnosed with EA with distal TEF (type C) receiving the repair surgery between June 2009 and January 2012 in our institution. Data collected included the newborns' gestational age and weight at the time of the operation, operative time, parameters of intraoperative mechanical ventilation, oxygenation, end-tidal carbon dioxide (ETCO2), and analysis of blood gases. Time to extubation and length of stay were also recorded.</p><p><b>RESULTS</b>Intravenous induction with muscle paralysis followed by pressure-control ventilation and tracheal intubation regardless of the position of the fistula can be performed uneventfully in EA/TEF newborns with no additional airway anomalies and large, pericarinal fistulas in our experiences. The thoracoscopic approach appeared to take longer than the open approach. During the procedure of repair, hypercarbia and acidosis developed immediately 1 hour after pneumothorax in both groups. CO2 insufflation did have additional influence on the respiratory function of the newborns in the TR group; values of PaCO2 and ETCO2 were higher in the TR group but the difference did not reach statistical significance. By the end of the procedure, values of PaCO2 and ETCO2 returned to the baseline levels while pH did not, but all parameters made no difference in the two groups. Besides, time to extubation was shorter in the TR group.</p><p><b>CONCLUSIONS</b>Thoracoscopic repair of EA/TEF is comparable to the open repair, and is believed to be safe and tolerable in selected patients. A wider range of neonates may be acceptable for thoracoscopic EA/TEF repair with increasing surgical experience.</p>

Construction of deltaNp63 specific small hairpin RNA expressing plasmid and its role in bladder cancer--a preliminary study / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To construct deltaNp63 specific small hairpin RNA (shRNA) expressing plasmid,to examine its inhibitory effect to the expression of deltaNp63 protein and mRNA in transitional cell carcinoma of the bladder (TCCB) , its effect on TCCB cells cycle and proliferation.</p><p><b>METHODS</b>DeltaNp63 specific oligonucleotides were designed and synthesized. These oligonucleotides were annealed to form double strand DNA fragments and this fragment was cloned into Pgenesil-1 plasmid. The recombinant deltaNp63-shRNA expression construct was confirmed by using Pst I + Sal I double digestion and by sequencing. Fluorescence staining was used to confirm the success of transfection in TCCB cells under the fluorescence microscope. The inhibitory effect of deltaNp63-shRNA construct was examined with semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical staining assay. The cell cycle of TCCB cells was assayed by flow cytometry (FCM). The cellular proliferation of TCCB cells was assayed by tetrazolium bromide (MTT) colorimetry.</p><p><b>RESULTS</b>The deltaNp63-shRNA expression plasmid was successfully constructed and transfected into TCCB cells. It can effectively reduce the expression of deltaNp63 protein and mRNA. The reduction rate of deltaNp63 mRNA was 63.0%, and the G0/G1 ratio was increased and S phase was decreased in transfected TCCB cells. The cellular proliferation was also lower in transfected 5637 cells in comparrison with that of non-transfected TCCB cells.</p><p><b>CONCLUSION</b>A deltaNp63-shRNA expression plasmid, constructed from Pgenesil-1 plasmid, can successfully be transfected into TCCB cells and can effectively inhibit the expression of deltaNp63 protein and mRNA. It also can take part in regulation of the cell cycling and inhibit the cellular proliferation of TCCB cells.</p>

Effect of saffor (Carthamus tinctorius) injection on renal ischemia/reperfusion injury in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect and mechanism of saffor injection on renal ischemia/reperfusion (I/R) injury in rats.</p><p><b>METHOD</b>Seventy-five SD rats were randomly divided into five groups (n = 15, in each), normal control groups, I/R control groups, low-dose treatment groups, middle-dose treatment groups and high-dose treatment groups. After rat's I/R injury model was established, renal function was assessed by measuring serum creatinine, blood urea nitrogen, urine osmotic pressure and urine osmotic pressure/blood osmotic pressure, the apoptosis rate in I/R renal tissure was measured by TUNEL method and caspase-3 concentration was measured by immunohistochemistry.</p><p><b>RESULT</b>Reperfusion of the ischemic kidney induced marked renal dysfunction. Saffor injection significantly inhibited the reperfusion-associated increase in apoptosis rate and caspase-3 protein absorbance value. Moreover, the renal dysfunction at all treatment groups was markedly ameliorated by Saffor injection. (P < 0.01).</p><p><b>CONCLUSION</b>The results show that saffor injection significantly reduces the renal dysfunction and injury caused by I/R of the kidney, And the protective effect of Saffor injection may be related to the inhibition of cell apoptosis and caspase-3 gene expression following renal I/R.</p>

Regulation of Tiangui and viscera function / 中华中医药杂志

Tiangui,a specific terms in TCM,is a special material that can regulate human reproduction and development. It stems from congenital kidney essence and is nourished by acquired nutrients with the nature of time-limit,rhythmicity and status. Tiangui promotes abundant CHONG meridian with unobstructed REN meridian,participates in the production and regulation of menstruation,and prepares for conception of women. According to the holism of TCM,person is a unified whole,Tiangui's generation and functional role must be completed under the condition of normal five viscera' function,especially the sufficient kidney essence,although the 'to'and 'exhaustion'of Tiangui is directly affected and regulated by congenital kidney essence. Therefore the synergistic effect of the five viscera can not be ignored on Tiangui,although the congenital kidney essence occupies an important position.