Objective To explore the effect of different packing contents of solid phase extraction column, repeated use times and solid phase extraction column under pressure on the determination of morphine content in compound liquorice oral solution. Methods The component was performed by high performance liquid chromatography （HPLC） equipped with the C₈ （4.6 mm × 150 mm, 5 μm） chromatographic column. The mobile phase consisted of acetonitrile-2.5 mmol/L sodium heptanesulfonate solution-50 mmol/L potassium dihydrogen phosphate solution （5∶18∶18）. The flow rate was 1.0 ml/min with the 220 nm detection wavelength, 20 μl injection volume and 30℃ column temperature. Results It was found that the SPE column produced of 500 mg had a higher recovery rate of morphine, and the RSD value was small. It was found that SPE column could be reused for up to 3 times under the condition of guaranteeing recovery rate.In the pressurized experiment,at the flow rate of 0.5 ml/min, the measured morphine content was basically consistent with that measured at atmospheric pressure. Conclusion The determination of morphine in compound liquorice oral solution could be carried out by C₁₈ solid phase extraction column of 500 mg. Under pressure, the flow rate was about 0.5 ml/min for morphine recovery, which could achieve high recovery of morphine in compound liquorice oral solution and improve the detection efficiency of morphine.

Objective:To investigate the efficacy of venetoclax-based combined regimen in treatment of adult patients with acute myeloid leukemia (AML).Methods:The data of 50 adult AML (non-acute promyelocytic leukemia) who received venetoclax-based combined regimen in the First Affiliated Hospital of Xiamen University, Dongguan People's Hospital, the First Hospital of Longyan City, Jieyang People's Hospital from December 2018 to May 2021 were retrospectively analyzed. Different doses venetoclax combined with demethylation drugs or low-dose chemotherapy regimen were used to analyze the therapeutic efficacy. The related factors influencing efficacy were analyzed by using logistic regression.Results:The composite complete remission (CR) rate of 50 AML patients was 62.0% (31/50), the overall response rate (ORR) was 76.0% (38/50); 28 patients achieved effectiveness [CR and partial remission (PR)] after the first cycle and could achieve effectiveness by 3 courses of treatment at the latest. Among 50 patients, 28 cases were newly diagnosed AML, the composite CR rate was 60.8% (17/28), ORR was 78.6% (22/28); 22 cases were recurrent and relapsed, the composite CR rate was 63.6% (14/22), ORR was 72.7% (16/22); and there was no statistically significant difference of ORR between the both groups ( χ2 = 0.23, P = 0.743). Logistic regression multivariate analysis showed age was the only independent influencing factor for the treatment effectiveness ( OR = 8.451, 95% CI 1.306-54.697, P = 0.025). The median duration time of patients receiving venetoclax treatment regimen was 4.5 months (1.1-15.0 months); 16 cases who had treatment effectiveness finally relapsed, the median time of maintaining effectiveness was 5 months (1.1-11.0 months). Additionally, the common treatment-related adverse reactions included bone marrow suppression after treatment, followed by some gastrointestinal reactions like nausea, vomiting and stomachache. In addition, no patient stopped medication for more than 1 week due to bone marrow suppression related complications. Conclusion:Venetoclax-based combined regimen shows a good short-term efficacy in treatment of AML. It is also effective and tolerable for elderly patients receiving reduced dose therapy.

Objective:To explore the influencing factors of the secondary sex ratio (SSR) of singleton babies born following assisted reproductive technology.Methods:A total of 7020 single babies delivered by assisted reproductive technology at the Reproductive Medicine Center of the First Affiliated Hospital of USTC (Anhui Provincial Hospital) from January 2000 to December 2018 were collected in a retrospective cohort study. The effect of influencing factors, such as maternal age, fertilization method, embryo transfer type and embryo transfer period on the sex of singleton babies were analyzed.Results:Among 7020 babies, 3730 were boys and 3290 were girls. The total SSR was 113∶100. The SSR of in vitro fertilization (IVF) was significantly higher than that of intracytoplasmic sperm injection (ICSI) fertilization (121∶100 vs. 95∶100, P<0.001), while other factors had no significant effect on the SSR of infants born. In the stratified analysis, there was no difference in SSR between IVF and ICSI in the blastocyst embryo transfer group with mother's age 35 years or older ( P<0.05), while the SSR of IVF in the other groups was higher than that of ICSI (<35 years old group: P<0.001; fresh embryo group: P=0.001; frozen-thawed embryo group: P=0.003; cleavage embryo transfer group: P<0.001). Univariate logistic regression analysis showed that the fertilization method in singleton pregnancy had a statistically significant effect on baby SSR ( OR=0.792, 95% CI=0.712-0.881, P<0.001). After removing confounding factors, logistic regression analysis showed that the fertilization method in singleton pregnancy had a statistically significant effect on neonatal SSR (a OR=0.793, 95% CI=0.713-0.883, P<0.001). Conclusion:The fertilization method of assisted reproductive technology can affect the SSR of singleton newborns. The SSR of IVF is significantly higher than that of ICSI and the difference was significant.

Objective:To explore the influencing factors of the secondary sex ratio (SSR) of singleton babies born following assisted reproductive technology.Methods:A total of 7020 single babies delivered by assisted reproductive technology at the Reproductive Medicine Center of the First Affiliated Hospital of USTC (Anhui Provincial Hospital) from January 2000 to December 2018 were collected in a retrospective cohort study. The effect of influencing factors, such as maternal age, fertilization method, embryo transfer type and embryo transfer period on the sex of singleton babies were analyzed.Results:Among 7020 babies, 3730 were boys and 3290 were girls. The total SSR was 113∶100. The SSR of in vitro fertilization (IVF) was significantly higher than that of intracytoplasmic sperm injection (ICSI) fertilization (121∶100 vs. 95∶100, P<0.001), while other factors had no significant effect on the SSR of infants born. In the stratified analysis, there was no difference in SSR between IVF and ICSI in the blastocyst embryo transfer group with mother's age 35 years or older ( P<0.05), while the SSR of IVF in the other groups was higher than that of ICSI (<35 years old group: P<0.001; fresh embryo group: P=0.001; frozen-thawed embryo group: P=0.003; cleavage embryo transfer group: P<0.001). Univariate logistic regression analysis showed that the fertilization method in singleton pregnancy had a statistically significant effect on baby SSR ( OR=0.792, 95% CI=0.712-0.881, P<0.001). After removing confounding factors, logistic regression analysis showed that the fertilization method in singleton pregnancy had a statistically significant effect on neonatal SSR (a OR=0.793, 95% CI=0.713-0.883, P<0.001). Conclusion:The fertilization method of assisted reproductive technology can affect the SSR of singleton newborns. The SSR of IVF is significantly higher than that of ICSI and the difference was significant.

Quantitative evaluation of analgesic efficacy improves understanding of the antinociceptive mechanisms of new analgesics and provides important guidance for their development. Lappaconitine (LA), a potent analgesic drug extracted from the root of natural Aconitum species, has been clinically used for years because of its effective analgesic and non-addictive properties. However, being limited to ethological experiments, previous studies have mainly investigated the analgesic effect of LA at the behavioral level, and the associated antinociceptive mechanisms are still unclear. In this study, electrocorticogram (ECoG) technology was used to investigate the analgesic effects of two homologous derivatives of LA, Lappaconitine hydrobromide (LAH) and Lappaconitine trifluoroacetate (LAF), on Sprague-Dawley rats subjected to nociceptive laser stimuli, and to further explore their antinociceptive mechanisms. We found that both LAH and LAF were effective in reducing pain, as manifested in the remarkable reduction of nocifensive behaviors and laser-evoked potentials (LEPs) amplitudes (N2 and P2 waves, and gamma-band oscillations), and significantly prolonged latencies of the LEP-N2/P2. These changes in LEPs reflect the similar antinociceptive mechanism of LAF and LAH, i.e., inhibition of the fast signaling pathways. In addition, there were no changes in the auditory-evoked potential (AEP-N1 component) before and after LAF or LAH treatment, suggesting that neither drug had a central anesthetic effect. Importantly, compared with LAH, LAF was superior in its effects on the magnitudes of gamma-band oscillations and the resting-state spectra, which may be associated with their differences in the octanol/water partition coefficient, degree of dissociation, toxicity, and glycine receptor regulation. Altogether, jointly applying nociceptive laser stimuli and ECoG recordings in rats, we provide solid neural evidence for the analgesic efficacy and antinociceptive mechanisms of derivatives of LA.
Aconitine/pharmacology* ; Analgesics/pharmacology* ; Animals ; Pharmaceutical Preparations ; Rats ; Rats, Sprague-Dawley

Quantitative evaluation of analgesic efficacy improves understanding of the antinociceptive mechanisms of new analgesics and provides important guidance for their development. Lappaconitine (LA), a potent analgesic drug extracted from the root of natural Aconitum species, has been clinically used for years because of its effective analgesic and non-addictive properties. However, being limited to ethological experiments, previous studies have mainly investigated the analgesic effect of LA at the behavioral level, and the associated antinociceptive mechanisms are still unclear. In this study, electrocorticogram (ECoG) technology was used to investigate the analgesic effects of two homologous derivatives of LA, Lappaconitine hydrobromide (LAH) and Lappaconitine trifluoroacetate (LAF), on Sprague-Dawley rats subjected to nociceptive laser stimuli, and to further explore their antinociceptive mechanisms. We found that both LAH and LAF were effective in reducing pain, as manifested in the remarkable reduction of nocifensive behaviors and laser-evoked potentials (LEPs) amplitudes (N2 and P2 waves, and gamma-band oscillations), and significantly prolonged latencies of the LEP-N2/P2. These changes in LEPs reflect the similar antinociceptive mechanism of LAF and LAH, i.e., inhibition of the fast signaling pathways. In addition, there were no changes in the auditory-evoked potential (AEP-N1 component) before and after LAF or LAH treatment, suggesting that neither drug had a central anesthetic effect. Importantly, compared with LAH, LAF was superior in its effects on the magnitudes of gamma-band oscillations and the resting-state spectra, which may be associated with their differences in the octanol/water partition coefficient, degree of dissociation, toxicity, and glycine receptor regulation. Altogether, jointly applying nociceptive laser stimuli and ECoG recordings in rats, we provide solid neural evidence for the analgesic efficacy and antinociceptive mechanisms of derivatives of LA.

Objective To observe the changes of short-chain acyl-CoA dehydrogenase (SCAD) expression on human umbilical vein endothelial cell (HUVEC) apoptosis and investigate its relationship with apoptosis. Methods The HUVEC was cultured normally for 2-3 days. The apoptotic model of HUVEC was established by tert-butyl hydrogen peroxide (tBHP). The HUVEC was treated by different concentrations of tBHP (0, 10, 20, 30, 40, 50 μmol/L) for 12 hours and different time (0, 3, 6, 9, 12 hours) with 50 μmol/L tBHP to establish the apoptotic model of HUVEC. The cell viability was detected by methyl thiazolyl tetrazolium (MTT), the mRNA expression of SCAD was determined by real-time polymerase chain reaction (PCR), the protein expression of SCAD was achieved by Western Blot. The best concentrate and time were determined to interfere the HUVEC to achieve the apoptotic model of HUVEC. The SCAD gene of HUVEC was knocked down by RNA interference sequence (siRNA274, siRNA414, siRNA679). The mRNA expression of SCAD, the protein expression of SCAD and the activity of SCAD enzyme were detected to achieve the best RNA interference sequence. The HUVEC was intervened by the best RNA interference sequence and tBHP. The cell activity and apoptosis rate, the enzyme activity of SCAD, the mRNA and protein expression of SCAD, the contents of reactive oxygen species (ROS), aderosine triphosphate (ATP) and free fatty acid (FFA) were detected to observe the effect of SCAD on apoptosis of HUVEC. Results ① The cell viability, the mRNA expression and the protein expression of SCAD were decreased gradually in a concentration and time dependent manner with the increase of tBHP concentration and the prolongation of intervention time. The decline was most significant in the group of the 50 μmol/L tBHP to interfere HUVEC for 12 hours. ② The siRNA679 transfection was the most significant in reducing SCAD mRNA and protein expressions among the three interference sequences (siRNA274, siRNA414, siRNA679). ③ Compare with blank control group, the cell viability was significantly decreased in the siRNA679 group (A value: 0.48±0.09 vs. 1.00±0.09, P < 0.01), the apoptotic rate of HUVEC was significantly increased [(29.96±2.09)% vs. (2.90±1.90)%, P < 0.01], the expression of SCAD mRNA and SCAD protein, the activity of SCAD enzyme and the content of ATP were significantly decreased [SCAD mRNA (2-ΔΔCt): 0.50±0.16 vs. 1.34±0.12, SCAD/α-Tubulin: 0.67±0.11 vs. 1.00±0.06, the activity of SCAD enzyme (kU/g): 0.38±0.04 vs. 0.53±0.04, the content of ATP (μmol/g): 0.14±0.02 vs. 0.19±0.01, all P < 0.05], the contents of FFA and ROS were significantly increased [FFA (nmol/g): 0.84±0.07 vs. 0.47±0.04, ROS (average fluorescence intensity): 647.5±23.7 vs. 434.2±46.5, both P < 0.01]. Meanwhile, SCAD siRNA treatment triggered the same apoptosis as HUVEC treated with tBHP. Conclusions Down-regulation of SCAD may play an important role in HUVEC apoptosis. Increase in the expression of SCAD may become an important part in intervening HUVEC apoptosis.

Objective To establish a method that could detect 5 components of Fufang Heishen oral liquid simultaneously. Methods The component was performed by high performance liquid chromatography ( HPLC) equipped with Agilent Hypersil ODS (4.6 mm×250 mm,5 μm).The mobile phase consisted of acetonitrile-0.1% Phosphoric acid with gradient elution.The flow rate was 1.0 mL·min-1with the 210 nm and 270 nm detection wavelength,20 μL injection volume and 30 ℃column temperature. Results A good linear relationship was observed with the range of 7.12-85.44 mg·L-1for Harpagide (r＝0.999 9),2.50-30.00 mg·L-1for Harpagoside(r＝0.999 8),25.35-304.20 mg·L-1for Cinnamic acid(r＝0.999 7),0.73-8.70 mg·L-1for Tectoridin(r＝0.999 7)and 1.20-14.40 mg·L-1for Irisflorentin(r＝0.999 8).The average recovery of each detected component of Fufang Heishen Oral Liquid was 98.8%,102.7%,98.8%,99.3%,99.9% the RSD were 1.23%,2.89%, 2.60%,1.44%,2.84%(n＝6). Conclusion The method is simple,rapid and accurate and can be used to detect the content of Harpagide,Harpagoside,Cinnamic acid,Tectoridin and Irisflorentin of Fufang Heishen Oral Liquid.

Objective To establish a quality standard for compound Heishen granules .Methods Scrophulariae Radix and Belamcandae Rhizoma were identified by TLC .HPLC was used to determine the content of cinnamic acid ,tectoridin and irisflo-rentin .The HPLC was performed on a column of Kromasil-C18 (150 mm × 4 .6 mm ,5 μm)with a mobile phase of acetonitrile (A)-0 .1％ hydrochloric acid (B)at a temperature 30 ℃ .The detection wavelength was set at 270 nm and the flow rate at 1 ml/min .Results The TLC method had good specificity without interference from negative control .The calibration curve showed a good linear relationship within the range of 16 .22-113 .57μg/ml for cinnamic acid(r=0 .9998) ,48 .19-337 .34μg /ml for tectoridin(r= 0.9998)and 16.40-114.80 μg/ml for irisflorentin(r= 0.9999) .The average recoveries were 99.23％ , 98.82％ ,99.17％ .Conclusion The established method is rapid ,accurate and reproducible .It can be used in the quality control of compound Heishen granules .

AIM:To observe the effect of metformin on the expression of GSTM2 in spontaneously hypertensive rats,and to investigate the mechanism of the reversal of myocardial hypertrophy by metformin.METHODS:Sixteen weeks old WistarKyoto (WKY) and spontaneously hypertensive rats (SHR) were used as the experimental control.Eight weeks old WKY and SHR were administered to metformin (Met) continuously for 8 weeks as the experimental group.Systolic pressure of rats was regularly determined by NIBP instrument of heart rate and blood pressure.The hemodynamic parameters were tested by BL-420 biological experimental system and 4 track physiological recorders.The weights of left ventricle and rats were collected to calculate the left ventricular mass index.The activity of GSTM2 enzyme in left ventricle was detected by Enzyme-linked immunosorbent assay (ELISA).The protein expression of GSTM2 and p47phox and Nox4 in left ventricle were investigated by Western blot.The O2·-levels were measured by staining with dihydroethidium (DHE).RESULTS:Compared with the experimental control,the blood pressure and left ventricular mass index and left ventricular end-diastolic pressure (LVEDP) increased in SHR;the maximal rate of increase and decrease of left-ventricle pressure development (± dp/dt max) decreased significantly in SHR;the protein expression and enzyme activity of left ventricular GSTM2 were significantly decreased;the expression of p47phox,Nox4 and the generation of O2·-were significantly increased in the left ventricles of SHR;the value of P was less than 0.01 and the difference was statistically significant.Compared with the SHR group,the SHR were administered with metformin (Met) continuously for 8 weeks.The left ventricular mass index and left ventricular end-diastolic pressure(LVEDP) decreased observably in SHR administration group;the maximal rate of increase and decrease of left-ventricle pressure development(± dp/dt max) increased in SHR administration group;the protein expression and enzyme activity of left ventricular GSTM2 were significantly increased in SHR administration group.the expression of p47phox,Nox4 and the generation of O2·-was significantly decreased in the left ventricles of SHR administration group;the value of P was less than 0.01 and the difference was statistically significant.CONCLUSION:Metformin can significantly reverse the myocardial hypertrophy in SHR,which might be associated with the up-regulation of GSTM2 expression,decrease the expression of p47phox,Nox4 and O2·-generation,elimination of oxidative stress.