ObjectiveTo establish the specific chromatogram and thin layer chromatography（TLC） of Qingxin Lianziyin（QXLZY） benchmark samples， in order to clarify the key quality attributes and provide a reference for the quality evaluation of QXLZY. MethodHigh performance liquid chromatography（HPLC） specific chromatogram of QXLZY benchmark samples was developed by using a YMC Hydrosphere C₁₈ column（4.6 mm×250 mm， 5 μm） with the mobile phase of acetonitrile（A）-0.2% formic acid aqueous solution（B） for gradient elution（0-10 min， 5%-20%A； 10-20 min， 20%A； 20-25 min， 20%-24%A； 25-40 min， 24%-30%A； 40-55 min， 30%-50%A； 55-65 min， 50%-100%A； 65-75 min， 100%A； 75-75.1 min， 100%-5%A； 75.1-90 min， 5%A）， and the detection wavelength was 360 nm. Ultra-high performance liquid chromatography-linear ion trap/orbitrap mass spectrometry（UHPLC-LTQ-Orbitrap MS） with electrospray ionization（ESI） was used to identify the components of QXLZY benchmark samples by accurate relative molecular weight and multilevel MS fragment ion information， the detection conditions were positive and negative ion modes and data dependency scanning mode. TLC identification methods for Ophiopogonis Radix， Lycii Cortex， Nelumbinis Semen， Poria， Astragali Radix and Ginseng Radix et Rhizoma in QXLZY were established. ResultA total of 15 characteristic peaks were identified from Glycyrrhizae Radix et Rhizoma， Plantaginis Semen and Scutellariae Radix， and the relative standard deviations of the retention times of 15 characteristic peaks in 15 batches of QXLZY benchmark samples were≤3% with peak 8（baicalin） as the reference peak. A total of 100 compounds， including flavonoids， organic acids， saponins， amino acids and others， were identified in the benchmark samples by UHPLC-LTQ-Orbitrap MS. The established TLC had good separation and was suitable for the identification of Ophiopogonis Radix， Lycii Cortex， Nelumbinis Semen， Poria， Astragali Radix and Ginseng Radix et Rhizoma in QXLZY. ConclusionThe material basis of QXLZY benchmark samples is basically determined by MS designation and source attribution. The established specific chromatogram and TLC of QXLZY are simple， stable and reproducible， which can provide a reference for the development and quality control of QXLZY.

Qingxin Lianzi Yin （QXLZY）， as an ancient classical formula for clearing the heart and nourishing the Yin， was composed of nine herbs （Scutellariae Radix， Ophiopogonis Radix， Lycii Cortex， Plantaginis Semen， Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle， Nelumbinis Semen， Poria， Astragali Radix and Ginseng Radix et Rhizoma）， coming from Prescriptions of the Bureau of Taiping People's Welfare Pharmacy. It could clear away the heart-fire， promote the interaction of the heart and kidney， replenish Qi and Yin， and stop strangury-turbidity. It was used to treat gonorrhea of urination， seminal emission， restlessness， wasting-thirst and so on. At present， the usage and dosage of QXLZY and its addition and subtraction are different in clinical practice. Most of the studies just focus on its clinical efficacy， and there is few review literature reflecting its historical evolution. Based on this， this paper systematically clarified the historical evolution， composition， preparation， interpretation， function， and modern clinical application of QXLZY. This work has been explained the historical evolution of QXLZY， and found that it was wildly used in modern clinical， especially suitable for the treatment of chronic urinary system diseases. At the same time， QXLZY also had significant therapeutic effects on neurasthenia， stomatitis， diabetic nephropathy and other aspects. Through the comprehensive analysis of ancient and modern literature， this work explores the true connotation of QXLZY from the perspective of traditional Chinese medicine theory， which can point out the direction of the clinical application and positioning of this famous classical formula after it comes into the market， and also can provide reference basis for its subsequent in-depth research and development.

To investigate the effects of ceramide pathway on the inhibition of artesunate (Art) to hepatic fibrosis. LX-2 cells were divided into control group, Art treated group with 350 μmol/L, fumonisin B1 (FB1) treated group with 6 μmol/L, and Co-administration group of artesunate 350 μmol/L and fumonisin B1 6 μmol/L. There were 7 compound holes in each group. After 24 hours of treatment, the cells and supernatant were collected and detected. The expressions of homo sapiens longevity assurance homologue 2 (LASS2), peroxisome proliferators-activated receptors-γ (PPAR-γ) and Caspase-3 were evaluated by Western blot, the content of ceramide was evaluated by HPLC-FLD method, MTT assay was adopted to measure the rate of proliferation of LX-2 cells. The content of hydroxyproline was determined by digestive method. Compared with the control group, the expression of ceramide synthase protein and the ceramide content were increased significantly, the proliferation of LX-2 cells was inhibited significantly, the expressions of PPAR-γ and Caspase-3 protein were up-regulated and the secretion of hydroxyproline was inhibited in Art treated group (P＜0.05). In FB1 treated group, the protein expression of ceramide synthase and the ceramide content were decreased significantly, the proliferation of LX-2 cells was increased significantly, the expressions of PPAR-γ and Caspase-3 protein were down-regulated, and the secretion of hydroxyproline was increased (P＜0.05). Compared with the Art alone group, the combination of the two drugs could significantly reduce the effects of Art on the expression of ceramide synthase protein and the increase of ceramide content, and attenuate the effects of Art on the cell proliferation , PPAR-γ, Caspase-3 protein expression and hydroxyproline level of LX-2 cells (P＜0.05). Artesunate could inhibit hepatic fibrosis by increasing the content of ceramide through the ceramide synthase-ceramide pathway.

Objective To compare and analyze the lower limb muscle activity and knee joint force during the stance periods of gait cycle in patients with osteoarthritis before and after total knee arthroplasty (TKA). Methods Based on the OpenSim platform, lower extremity musculoskeletal models of one healthy subject and three patients with osteoarthritis before and after TKA were established. A three-dimensional （3D） motion capture system and a force platform were used to collect the lower limb kinematic data and the ground reaction force during walking, which were used as input parameters to simulate the lower limb muscle activation and knee joint forces. Results The results from the musculoskeletal model were consistent with the results by inverse dynamics based on the 3D motion capture system. The patient's rectus femoris was activated in the loading response and mid stance phases, which was different from that of healthy subject. The activation timing and amplitude of the quadriceps muscle in 3 patients were significantly different before and after TKA. The peak joint forces of 3 patients before TKA were 2.95, 3.15 and 3.43 times of body weight (BW) with the constant load of more than 2 times of BW during stand phase. The peak joint force after TKA were 2.09, 2.48 and 3.96 times of BW respectively. The joint force was not improved and the knee function did not reach the normal level six months after TKA. Conclusions The results of the established musculoskeletal model have certain reliability, and this model can provide a biomechanical auxiliary method for TKA surgery in the future.

Objective: To determine whether intrauterine infection with hepatitis B virus (HBV) occurs in early pregnancy and to characterize associated virulence factors. Methods: Villi tissues and blood samples of 45 HBV surface antigen (HBsAg)-positive pregnant women were collected during the first trimester and HBV DNA loads were quantified by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The expression of GCM1, HBsAg and hepatitis B core antigen (HBcAg) in villi tissues were detected by immunohistochemical method. Results: Data from qRT-PCR showed that HBV DNA was detected in 14 of 45 villi tissues (positive rate of 31.11%), and 24 of 45 blood samples (positive rate of 53.33%), further statistical analysis showed that the positive rates of HBV DNA between blood samples and villi tissues were not significantly different (χ²=4.555, P=0.054). Among them, 12 samples were consistently positive between the villi and blood specimens, and HBsAg, HBeAg, HBeAb, HBV DNA from peripheral blood in these pregnant women were significantly higher than those of the other women (P value was 0.007, 0.004, 0.000, and 0.000 respectively). The multivariate logistic regression analysis showed that blood HBV DNA greater than 106 IU/ml was independently associated with HBV DNA positive in villi, and the HBsAg, HBeAg, villi tissues HBV DNA positive rates of these pregnant women were significantly higher than those of the other pregnant women (all P value were 0.000). Immunohistochemistry results showed that all 45 cases were positive for GCM1 expression in the cell nucleus. Nine cases also had HBsAg expression in the cytoplasm. Only one case was found to express HBV core antigen (HBcAg) in the nucleus. Conclusions HBV DNA and HBsAg can be detected from villi tissues harvested during the first trimester in HBsAg-positive pregnant women, and the results suggest an early occurrence of intrauterine infection of fetuses with high HBV levels.

Medical ethics has a long history and rich connotations.It has developed from the simple "medical morality" of ancient times to the modem medical ethics.The basic principles of medical ethics include autonomy,non-maleficence,beneficence,justice,and so on.Researchers often conduct clinical researches in the balance between achievements and ethical norms.Clinical researchers of surgery should have a deep understanding of medical ethical principles and strictly abide by medical ethics.Ethics committee should strictly perform their duties and play the role of inspection and supervision.Modem medical knowledges should be popularized throughout the society to make clinical research correctly understood.Adhering principles of ethics first,people orientation and cooperation practice,with patients' benefit as evaluation criteria,balance of surgical "Dao" and "Shu" can be achieved.

Aims To study the effects of clenbuterol on anoxia/reoxygenation( A/R) injury in neonatal Wistar rat cardiomyocytes and to explore whether its mecha-nism is related to reperfusion injury salvage kinase ( RISK) or not. Methods The cultured primary neo-natal cardiomyocytes were randomly divided into eight groups: ①normal culture group; ②anoxia/reoxygen-ation( A/R) group;③ clenbuterol ( 1 μmol · L-1 ) +A/R;④ICI118,551(10 μmol·L-1) + clenbuterol ( 1 μmol · L-1 ) + A/R; ⑤Metoprolol ( 10μmol · L-1 ) + clenbuterol(1μmol·L-1 ) + A/R group;⑥Metoprolol ( 10 μmol · L-1 ) + A/R group; ⑦PD98059 ( 20 μmol · L-1 ) + clenbuterol ( 1 μmol · L-1 ) + A/R group;⑧ LY294002(10 μmol·L-1 ) +clenbuterol(1 μmol · L-1 ) + A/R group. Cell via-bility was determined by the conventional MTT reduc-tion assay. The content of LDH in cultured medium was measured with colorimetry. Cardiomyocyte apopto-sis was determined by Hoechst33342 . Intracellular re-active species( ROS) were monitored by the fluorescent DCFH-DA. Total ERK2 and phosphorylated ERK were detected by western blot. Results Compared with A/R group, clenbuterol significantly increased vaibility of cells, reduced LDH release, lowered the rate of apop-tosis and ROS production. When addedβ2 receptor an-tagonist ICI118 , 551 , PI3 K inhibitor LY294002 and ERK inhibitor PD98059 , the effects of clenbuterol a-bove were inhibited; but β1 receptor antagonist Meto-prolol protected the cardiomyocytes from A/R injury, as evidenced by decreased LDH release and increased cell viability. There were no synergistic effects in the combined use of clenbuterol and Metoprolol. Conclu-sion clenbuterol exerts cardioprotective effects against A/R injury by inhibiting oxidative stress and apopto-sis. The protection of clenbuterol is inhibited by ICI118 , 551 , LY294002 and PD98059 . clenbuterol protects cardiomyocytes against A/R injury via RISK pathway by activation of β2 receptor.

OBJECTIVE: To investigate the expression and clinical significance of BUB1 protein in laryngeal squa mous cell carcinoma (LSCC) tissues. METHOD: The expression of BUB1 protein was measured by immunohistochemistry (EliVision two steps) in 55 cases of LSCC tissues,30 cases of adjacent normal laryngeal mucosa tissues, the relationship of BUB1 expression with clinical pathologic factors in LSCC was also analyzed. RESULT: The positive rate of BUB1 protein in LSCC tissues (50.9%, 28/55) was significantly lower than that in adjacent normal laryngeal mucosa tissues (83.3%, 25/30), P < 0.01. The positive rate of BUB1 protein was correlated with differentiation grading and lymph node metastasis of LSCC (P < 0.05, respectively), but not with patient's sex, age, smoking, tumor site, T-stage and clinical stage(P > 0.05, respectively). CONCLUSION The low expresssion of BUB1 protein is closely related to the tumorigenesis and development of LSCC, and can be a molecular marker for prognosticating metastasis and prognosis of LSCC.
Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Laryngeal Mucosa ; metabolism ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; Prognosis ; Protein-Serine-Threonine Kinases ; metabolism

Objective To measure the pressure-volume changes and uroflow rate,and assess the function of upper urinary tract in sigmoidorcctal pouch patients.Methods Between 2007 and 2011,a to-tal of 10 patients who had a radical cystectomy because of muscle-invasive bladder tumor underwent a Mainz pouch Ⅱ procedure were evaluated.The ureters were implanted into the post walls of the detubularized sigmoid segment at least 30 cm in length.In all cases,sigmoidoscopy was done and the anal pressure was measured preoperatively,and video urodynamic study of sigmoidorectal pouch as well as the intravenous pyelography was conducted after 3 to 6 months postoperatively.Results Video urodynamic study of sigmoidorectal pouch revealed that no reflux up to an average volume of 360 ml (270-532 ml) of the descending colon.The sigmoidal colon pressure was 26 cm H2O during the 3 to 6 months post-operative follow-up.The anal sphincter pressure was 90 cm H2O on average preoperatively and did not change postoperatively.The renal function and upper urinary tract were preserved well.The Qmax was 30 ml/s and the average uroflow rate was 8 ml/s under abdominal strain.All the patients suffered slight incontinence in the first two months and became continent since the 3 month postoperatively with nocturnal voiding one to four times.Conclusions The sigmoidorectal pouch provides a reservoir with a higher capacity,lower pressure without reflux to the upper urinary tract and descending colon and lower metabolic acidosis problem.It is also a good alternative diversion procedure that would preserve upper urinary tract and good quality of life.

The different effects of capsaicin on I(A) and I(K) currents in pain-conduct neurons of trigeminal ganglia (TG) were investigated. In cultured TG neurons of rats, whole-cell patch clamp techniques were used to record the I(A) and I(K) before and after capsaicin perfused. Results revealed that 1 micromol/L capsaicin could inhibit the amplitude of I(A) by 48.2% (n = 10, P < 0.05), but had no inhibitory effect on I(K) (n = 7, P > 0.05). Ten micromol/L capsaicin could significantly inhibit the amplitude of I(A) by 93.2% (n = 8, P < 0.01), but only slightly inhibit the amplitude of I(K) by 13.2% (n = 7, P < 0.05). Neither 1 micromol/L nor 10 micromol/L capsaicin had effects on the active curve of I(A) and I(K). It was concluded that capsaicin could selectively inhibit the I(A) current, and this effect might involve in the analgesic mechanisms of capsaicin.