Effect of clonal hematopoiesis in remission on hematopoiesis recovery in patients with NPM1 mutated acute myeloid leukemia after chemotherapy / 中华血液学杂志

Objective: To investigate the effect of clonal hematopoiesis (CH) in remission on hematopoiesis recovery in patients with NPM1 mutated acute myeloid leukemia (AML) after chemotherapy. Methods: Retrospective analysis was performed on 86 patients with NPM1(mut) AML newly diagnosed and treated in the First Affiliated Hospital of Soochow University between July 2016 and June 2019. Their clinical data and NGS test results at diagnosis were analyzed. Moreover, bone marrow samples in remission were tested using Sanger sequencing. The log-rank test was used to analyze the difference in hematopoietic recovery, and Cox proportional hazard models were used to analyze the prognostic factors affecting hematopoietic recovery. Results: The median age of the 86 NPM1(mut) AML patients was 50 years (15-69 years). There were 39 males and 47 females. Forty-one patients were induced with intensity chemotherapy ("7 + 3"), whereas 45 patients were treated with low-dose cytarabine-based induction chemotherapy. At diagnosis, The most common mutations in the patients were FLT3, DNMT3A, TET2, and IDH1/IDH2 mutations. CH-associated mutations persisted in 21 patients during remission, and the mutations were DNMT3A, TET2, ASXL1, and IDH1/IDH2. The recovery time of neutrophils in patients with CH-associated mutations in remission was consistent with that in patients without CH in remission (P=0.282) but the recovery time of platelets in patients with CH in remission was significantly longer[26 (95% CI 21-32) days vs 25 (95% CI 23-26) days, P=0.032]. Furthermore, univariate analysis indicated that age, induced chemotherapy program, and CH in remission were risk factors for platelet recovery, whereas multivariate analysis indicated that induced chemotherapy program and CH in remission were independent risk factors for platelet recovery (HR=0.454, P=0.001 and HR=0.520, P=0.027, respectively) . Conclusion: CH in remission delays the hematopoietic recovery of patients with NPM1(mut) AML after chemotherapy.

Analysis of clinical significance and prognostic impact of TET2 single nucleotide polymorphism I1762V in patients with acute myeloid leukemia / 中华血液学杂志

Objective: This study aimed to investigate the clinical and prognostic significance of TET2 single nucleotide polymorphism I1762V in patients with acute myeloid leukemia (AML) . Methods: The high-throughput sequencing method was used to sequence 58 hematological tumor-related genes in bone marrow samples from 413 patients with AML. TET2 I1762V and other somatic mutations were annotated and compared with patients' clinical information and prognosis. Results: I1762V was found in 154 patients with AML, which was significantly different from the general population in NyuWa Chinese Population Variant Database (χ(2)=72.4, P<0.001) . I1762V was not related to sex, age, and karyotype of patients with AML (P>0.05) . Patients with I1762V had a significantly higher proportion of NPM1 and KIT gene mutations than others (P<0.001) . NPM1 and KIT mutations were mutually exclusive. The survival analysis results revealed that the overall survival (OS) and progression-free survival (PFS) of patients with AML with I1762V were significantly greater than those of wild-type patients (HR=0.57, P=0.030; HR=0.55, P=0.020) , whereas the OS and PFS in patients with AML with DNMT3A mutation (with or without I1762V mutation) were lower than those of wild-type patients (HR=1.79, P=0.030; HR=1.74, P=0.040) . Conclusion: TET2 SNP I1762V has been linked to AML. I1762V is a prognostic factor of patients with AML, which can be used to guide the treatment and evaluate the prognosis of AML.

Inferring Postmortem Submersion Interval in Rats Found in Water Based on Vitreous Humor Metabolites / 法医学杂志

OBJECTIVES@#The metabolomics technique of LC-MS/MS combined with data analysis was used to detect changes and differences in metabolic profiles in the vitreous humor of early rat carcasses found in water, and to explore the feasibility of its use for early postmortem submersion interval (PMSI) estimation and the cause of death determination.@*METHODS@#The experimental model was established in natural lake water with 100 SD rats were randomly divided into a drowning group (n=50) and a postmortem (CO2 suffocation) immediately submersion group (n=50). Vitreous humor was extracted from 10 rats in each group at 0, 6, 12, 18 and 24 h postmortem for metabolomics analyses, of which 8 were used as the training set to build the model, and 2 were used as test set. PCA and PLS multivariate statistical analysis were performed to explore the differences in metabolic profiles among PMSI and causes of death in the training set samples. Then random forest (RF) algorithm was used to screen several biomarkers to establish a model.@*RESULTS@#PCA and PLS analysis showed that the metabolic profiles had time regularity, but no differences were found among different causes of death. Thirteen small molecule biomarkers with good temporal correlation were selected by RF algorithm. A simple PMSI estimation model was constructed based on this indicator set, and the data of the test samples showed the mean absolute error (MAE) of the model was 0.847 h.@*CONCLUSIONS@#The 13 metabolic markers screened in the vitreous humor of rat corpses in water had good correlations with the early PMSI. The simplified PMSI estimation model constructed by RF can be used to estimate the PMSI. Additionally, the metabolic profiles of vitreous humor cannot be used for early identification of cause of death in water carcasses.

Protopine inhibits the growth of hepatocellular carcinoma through a mitochondrially mediated apoptosis pathway / 药学学报

Hepatocellular carcinoma (HCC) is a serious threat for human health, the incidence of HCC in China accounts for more than 50% worldwide. There is an urgent need to develop novel anticancer agents for the treatment of HCC patients. Here we characterized the inhibitory effect and the molecular mechanism of protopine on HCC cancer cells. The results of a CCK-8 assay indicated that protopine displays anticancer activities on HCC cells. Flow cytometry and JC-1 staining confirmed that treatment with protopine decreased the mitochondrial membrane potential and induced apoptosis in HCC cells. Western blot analysis showed that protopine was able to increase protein expression in the mitochondrial apoptotic pathway; the level of cytochrome C, apoptotic protease activating factor-1 (Apaf-1), Bax, cleaved-poly ADP-ribose polymerase (cleaved-PARP), cleaved-caspase-3, and cleaved-caspase-9 were increased while the expression of Bcl-2 was suppressed significantly. An in vivo study revealed that protopine significantly suppressed the growth of tumors in nude mice bearing HepG2 cells. Administration of protopine intraperitoneally at a concentration of 50 mg·kg-1 inhibited tumor growth by 72.46%. Animal experiments were carried out according to the Regulation of the Animal Ethics Committee of Southwest Medical University. This study provides preliminary evidence that there is potential to develop protopine as a lead compound for the treatment of HCC.

Application Prospect of MicroRNA in Skin Wound Age Estimation / 法医学杂志

Wound age estimation is one of the major tasks in forensic practice. However, relatively accurate estimation of the wound age is still a conundrum and research spotlight world-widely. Studies show that microRNAs (miRNAs) are involved in the whole process of the skin wound repair, and miRNAs, as biomarkers, might be used to estimate the time of skin injury owing to their characteristic advantage. This paper summarizes the miRNA fundamental function, properties, current research progress in the estimation of wound age, and its limitations, and put forward prospect of potential application and research based on miRNAs in estimation of wound age.

Acupuncture-moxibustion treatment by stages based on the theory of "transformation of / 中国针灸

OBJECTIVE@#To compare the efficacy between acupuncture-moxibustion treatment by stages and femoston for premature ovarian insufficiency (POI).@*METHODS@#A total of 66 patients with POI were randomly divided into an observation group (33 cases, 3 cases dropped off) and a control group (33 cases, 2 cases dropped off). The patients in the observation group, based on the theory of "transformation of @*RESULTS@#Compared before treatment, the serum levels of FSH and LH were decreased (@*CONCLUSION@#Acupuncture- moxibustion treatment by stages based on the theory of "transformation of

Observation on therapeutic effect of electroacpuncture combined with penetrating moxibustion for postpartum pelvic organ prolapse / 中国针灸

OBJECTIVE@#To compare the clinical therapeutic effect of electroacupuncture (EA) combined with penetrating moxibustion and biofeedback electrical stimulation on postpartum pelvic organ prolapsed (POP).@*METHODS@#A total of 60 patients with POP who had delivery 6 weeks ago were randomized into an observation group and a control group, 30 cases in each one. In the observation group, EA was applied at Zigong (EX-CA 1), Ciliao (BL 32), Huiyang (BL 35), etc. while penetrating moxibustion was performed at acupoints of abdomen and lumbosacral region alternately every other day. In the control group, biofeedback electrical stimulation was provided. The treatment for 6 weeks was given once every other day, 3 times a week in both groups. Before treatment, after treatment and 6 months after delivery, pelvic floor muscle strength, pelvic organ prolapse quantification (POP-Q) evaluation and pelvic floor impact questionnaire short form-7 (PFIQ-7) were observed to assess the therapeutic effect.@*RESULTS@#Compared before treatment, the sustained contraction and rapid contraction force of pelvic floor muscle after treatment and 6 months after delivery were increased in both of the two groups (<0.05), and the changes in the observation group were larger than those in the control group (<0.05). After treatment and 6 months after delivery, the POP degree in the observation group was alleviated to the control group (<0.05). Compared before treatment, the scores of PFIQ-7 after treatment and 6 months after delivery were reduced in the two groups (<0.05), and the changes in the observation group were larger than those in the control group (<0.05).@*CONCLUSION@#Electroacupuncture combined with penetrating moxibustion can strengthen the pelvic floor muscle contractility of patients with postpartum pelvic organ prolapse, and are superior to biofeedback electrical stimulation in improving the pelvic organ prolapse status and life quality.

Research Progress on Estimation of Postmortem Submersion Interval / 法医学杂志

Postmortem interval （PMI） estimation is one of the most important and difficult academic tasks in forensic sciences. Due to the influence of the corpse itself and the water environment, corpses in water have unique corruption phenomenon and laws. Based on the experience of traditional PMI studies of corpses on land, forensic practitioners across the world have proposed a variety of practical methods for estimating postmortem submersion interval （PMSI）. This paper summarizes the literatures related to PMSI in recent years, and introduces methods to infer PMSI according to the phenomenon of corpses, the development of insects, the succession pattern of aquatic organisms, and the changes of other physical and chemical indexes of corpses, in order to provide some reference for the study of PMSI of corpses in water.

RGFP966 inactivation of the YAP pathway attenuates cardiac dysfunction induced by prolonged hypothermic preservation / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Oxidative stress and apoptosis are the key factors that limit the hypothermic preservation time of donor hearts to within 4-6 h. The aim of this study was to investigate whether the histone deacetylase 3 (HDAC3) inhibitor RGFP966 could protect against cardiac injury induced by prolonged hypothermic preservation. Rat hearts were hypothermically preserved in Celsior solution with or without RGFP966 for 12 h followed by 60 min of reperfusion. Hemodynamic parameters during reperfusion were evaluated. The expression and phosphorylation levels of mammalian STE20-like kinase-1 (Mst1) and Yes-associated protein (YAP) were determined by western blotting. Cell apoptosis was measured by the terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) method. Addition of RGFP966 in Celsior solution significantly inhibited cardiac dysfunction induced by hypothermic preservation. RGFP966 inhibited the hypothermic preservation-induced increase of the phosphorylated (p)-Mst1/Mst1 and p-YAP/YAP ratios, prevented a reduction in total YAP protein expression, and increased the nuclear YAP protein level. Verteporfin (VP), a small molecular inhibitor of YAP-transcriptional enhanced associate domain (TEAD) interaction, partially abolished the protective effect of RGFP966 on cardiac function, and reduced lactate dehydrogenase activity and malondialdehyde content. RGFP966 increased superoxide dismutase, catalase, and glutathione peroxidase gene and protein expression, which was abolished by VP. RGFP966 inhibited hypothermic preservation-induced overexpression of B-cell lymphoma protein 2 (Bcl-2)-associated X (Bax) and cleaved caspase-3, increased Bcl-2 mRNA and protein expression, and reduced cardiomyocyte apoptosis. The antioxidant and anti-apoptotic effects of RGFP966 were cancelled by VP. The results suggest that supplementation of Celsior solution with RGFP966 attenuated prolonged hypothermic preservation-induced cardiac dysfunction. The mechanism may involve inhibition of oxidative stress and apoptosis via inactivation of the YAP pathway.

[Role of nitric oxide in iron-induced toxicity in rat hearts].

The aim of the present study was to explore the effect of nitric oxide (NO) on iron-induced toxicity in rat hearts. Langendorff perfused rat heart and enzymatically isolated cardiomyocytes were used. It was shown that lipophilic Fe-HQ reduced the contractile amplitude, velocity and end-diastolic cell length in the cardiomyocyte, while the left ventricular developed pressure (LVDP), +/-dp/dt(max), heart rate and coronary flow showed biphasic alterations, which increased in the first 2 min and then was followed by a decline in isolated perfused rat heart; the contents of lactate dehydrogenase (LDH) and creatine kinase (CK) in the coronary effluent and the malondialdehyde (MDA) in the myocardium were increased. L-arginine (L-Arg), an NO precursor, reduced the contractile amplitude and end-diastolic cell length in the cardiomyocyte; but reversibly increased LVDP, +/-dp/dt(max), and coronary flow in isolated perfused rat heart. Pretreatment with L-Arg aggravated the Fe-HQ-induced decrease in contractile amplitude, velocity and end-diastolic cell length in the cardiomyocyte; LVDP, +/-dp/dt(max), heart rate and coronary flow were significantly reduced in the perfused heart, and the levels of LDH and CK increased in the coronary effluent. In contrast, the NOS inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) blocked the Fe-HQ induced change in contractile amplitude, velocity and end-diastolic cell length in the cardio- myocyte; it inhibited the decrease in LVDP, LVEDP and +/-dp/dt(max), and reduced the LDH and CK. Removing endothelial cells in coronary vessels attenuated the increase in LVDP and +/-dp/dt(max) at the beginning of Fe-HQ perfusion. It is suggested that L-Arg aggravates the iron-induced cardiac dysfunction, NO can mediate the iron-induced toxicity in heart, and endothelial cells in coronary vessels play an important role in the early stage of the effect of iron.

Efficacy of electroacupuncture combined with penetrating moxibustion for postpartum stress urinary incontinence / 中国针灸

OBJECTIVE@#To compare the clinical effect differences between electroacupuncture (EA) combined with penetrating moxibustion and the biological feedback training of pelvic floor muscle for postpartum stress urinary incontinence (SUI).@*METHODS@#Sixty patients of SUI who had delivery 42 days ago were randomly divided into an observation group and a control group, 30 cases in each one. The observation group was treated with EA and penetrating moxibustion. EA was applied at Ciliao (BL 32) and Huiyang (BL 35), combined with acupuncture at Qihai (CV 6), Zhongji (CV 3), Zigong (EX-CA 1), Zusanli (ST 36) and Sanyinjiao (SP 6); penetrating moxibustion was performed on abdomen and lumbosacral area. The control group was treated with biological feedback training of pelvic floor muscle. Both the groups were treated once every other day, 3 times per week for continuous 6 weeks. The International Consultation on Incontinence Questionnaire-Short Form (ICI-Q-SF), 1 h urinal pad test and pelvic floor muscle strength were tested before and after treatment; the efficacy was evaluated after treatment and at 6-month follow-up visit.@*RESULTS@#Compared before treatment, the ICI-Q-SF score and 1 h urine leakage were significantly reduced after treatment in the two groups (0.05). After treatment, the cured rate and total effective rate were 70.0% (21/30) and 96.7% (29/30) in the observation group, which were superior to 33.3% (10/30) and 70.0% (21/30) in the control group (<0.01); in the 6-month postpartum period, the cured rate and total effective rate were 63.3% (19/30) and 93.3% (28/30) in the observation group, which were superior to 30.0% (9/30) and 66.7% (20/30) in the control group (<0.05).@*CONCLUSION@#EA combined with penetrating moxibustion could improve the urinary control ability, relieve the symptoms of urinary incontinence and have a better long-term effect in patients with postpartum SUI, which is superior to biological feedback training of pelvic floor muscle.

Research Progress?on?Age?Determination?of?Venous?Thrombosis in Forensic Medicine / 法医学杂志

Pulmonary thromboembolism （PTE）, which is caused by detachment of venous thrombosis, is a common cause of sudden death in forensic practice. In the cases which die of PTE after trauma or die of PTE during non-thrombosis disease hospitalization, forensic pathologists are required to analyze the time sequence between trauma or medical practice and venous thrombosis, and then analyze their causal relationship. This review summarizes the history of thrombus age estimation and recent advances in forensic medicine, and then gives a brief outlook for future research to provide reference for forensic identification of PTE and guide follow-up studies.

Expression of Nrf2 in Different Cells after Human Cerebral Cortex Contusion / 法医学杂志

Objective To observe the expression changes of nuclear factor-erythroid derived 2-related factors （Nrf2） in different cells at different time points after human cerebral cortex contusion, and to discuss its application in brain wound age estimation. Methods Thirty-six human brain tissues were selected, of which 6 were for control and 30 were cortical contusion at different time points post-injury, which were divided into 0-1 h, 3-6 h, 1-3 d, 5-7 d, and 10-14 d post-injury groups, with 6 cases in each group. Based on paraffin embedded sections, HE staining was used to observe the morphological changes post-injury, and double immunofluorescence staining was used to detect the expression of Nrf2 in neurons, astrocytes, and microglia. The number of positive cells was counted and statistical analysis was made. Results The number of neurons decreased 1-3 d post-injury. The expression of Nrf2 cells in neurons increased after injury, and the rate of positive cells peaked at 1-3 d post-injury. Glial cells were activated 1-3 d post-injury, and the activation peaked at 5-7 d post-injury. The cerebromalacia began to form at 10-14 d post-injury. Glial fibrillary acidic protein （GFAP） positive cells in mice increased gradually after injury and peaked at 5-7 d post-injury, while the proportion of Nrf2 in GFAP positive cells was relatively stable. After injury, ionized calcium-binding adapter molecule 1 （IBA1） positive cells increased and activated gradually. The expression proportion of Nrf2 in IBA1 positive cells increased gradually, reached its peak at 5-7 d post-injury, and then decreased. Conclusion The expression of Nrf2 in different cells involves in the biological function of different cells post-injury, and the dynamic expression of single cells has a time-dependent pattern. This may provide a new reference index for the wound age estimation of brain contusion in human.

The Expression Changes of JNK3 and APP Protein in Apoptosis of Retinal Ganglion Cells after Optic Nerve Injury / 昆明医科大学学报

Objective To investigate the changes in the expression of JNK3 and APP protein in retinal ganglion cells (RGCs) after optic nerve injury and its effect on the survival of RGCs. Methods Optic nerve crush models were established using adult APP wild type mice. Immunohistochemistry and western blot for detecting the expression of JNK3 and APP protein in RGCs were used post-injury. The number of RGCs in APP KO mice group was counted to compare with APP WT mice group. Results After optic nerve injury, JNK3 expressed in cytoplasm of RGCs; p-JNK transformed from cytoplasm to nuclei of RGCs; APP expressed in cytomembrane; p-APP expressed in cytoplasm and axons of RGCs. Western blot results showed that the protein expressions of JNK3, p-JNK, APP and p-APP were increased in the injured group (P＜0.01) . The survival number of RGCs (147± 7.0) in APP KO mice group increased significantly than APP WT mice group (127.7 ±7.0) (P＜0.05) . Conclusion The upregulation of JNK3 and APP protein expression maybe play an important role in RGCs survival proceeds after optic nerve injury.

Relationship between Tissue Distributions of Modified Wuzi Yanzong Prescription () in Rats and Meridian Tropism Theory / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between tissue distributions of modified Wuzi Yanzong prescription (, MWP) in rats and meridian tropism theory.</p><p><b>METHODS</b>A high-performance liquid chromatography with Fourier transform-mass spectrometry (HPLC-FT) method was used to identify the metabolites of MWP in different tissues of rats after continued oral administration of MWP for 7 days. The relationship between MWP and meridian tropism theory was studied according to the tissue distributions of the metabolites of MWP in rats and the relevant literature.</p><p><b>RESULTS</b>Nineteen metabolites, mainly flavanoid compounds, were detected in the different rat tissues and classified to each herb in MWP. Further, it was able to establish that the tissue distributions of the metabolites of MWP were consistent with the descriptions of meridian tropism of MWP available in literature, this result might be useful in clarifying the mechanism of MWP on meridian tropism. In the long run, these data might provide scientific evidence of the meridian tropism theory to further promote the reasonable, effective utilization, and modernization of Chinese medicine.</p><p><b>CONCLUSION</b>The tissue distributions of MWP in vivo were consistent with the descriptions of meridian tropism of MWP.</p>

Relationship of Methylation within Upper Stream Region of Transcription Starts Site of HOXA5 Gene with Neural Tube Defects / 中国医学科学院学报

Objective To investigate the relationship between the methylation level of transcription starts site (TSS) upper stream of homeobox gene and the neural tube defects (NTDs). Methods A case-control study of two stages was designed. In the first stage,10 cases and 8 controls were extracted,in whom Illumina Infinium Human Methylation 450 k genome-wide beadchip was used for the quantification of DNA methylation levels of brain and spinal tissue. In the second stage,differentially methylated region within HOXA5 gene was detected with a larger numbers of samples (52 cases and 23 controls). DNA of brain or spinal tissue was extracted,and Matrix-assisted laser desorption ionization time-of-flight mass spectrometry technique of MassARRAY platform was employed for the validation of differentially methylated region of HOXA5 gene. Results In the first stage,27 CpG sites within TSS region of HOXA5 gene were found to be significantly hyper-methylated in case group compared to control group (P<0.05). In the second stage,a total of 10 CpG sites were analyzable within the differentially methylated region in the first stage. In the NTD case group,spinal bifida subgroup,and anencephaly subgroup,there were 7,6,and 2 sites with significantly higher methylation levels than that of control group (P<0.05). The average methylation level of TSS upper stream region within HOXA5 gene was higher in case group than control group [case group:(31.3±13.9)%,control group:(21.4±9.7)%],and the odds ratio after adjusting gender of fetus and pregnant week was 1.09 (1.03-1.16). Conclusion Hypermethylation within TSS upper stream region of HOXA5 gene in fetus is associated with a higher risk of NTDs.

Progress of research on the treatment of diabetic nephropathy with renin angiotensin aldosterone inhibitors / 中国生化药物杂志

Diabetes is one of the most common clinical chronic diseases, the incidence can lead to a variety of complications, diabetic nephropathy is one of them, because the incidence of diabetes is increasing year by year, so the incidence of diabetic nephropathy also gradually upgrade. The activation of the renin-angiotensin-aldosterone system (RAAS) is important in the occurrence and progression of the disease. RAAS inhibitors have a good control over the patient's blood pressure and can also be reduced in urinary protein small ball high filtration degree decreased, its renal interstitial fibrosis also has a certain inhibitory effect. The study showed that the RAAS inhibitor slowed the progression of the disease, the incidence of end-stage renal disease decreased significantly. This article will elaborate and analyze the progress of the application of RAAS inhibitors in the treatment of diabetic nephropathy to help more researchers enhance their understanding and improve the clinical efficacy of diabetic nephropathy.

Progress of research on the treatment of diabetic nephropathy with renin angiotensin aldosterone inhibitors / 中国生化药物杂志

Diabetes is one of the most common clinical chronic diseases, the incidence can lead to a variety of complications, diabetic nephropathy is one of them, because the incidence of diabetes is increasing year by year, so the incidence of diabetic nephropathy also gradually upgrade. The activation of the renin-angiotensin-aldosterone system (RAAS) is important in the occurrence and progression of the disease. RAAS inhibitors have a good control over the patient's blood pressure and can also be reduced in urinary protein small ball high filtration degree decreased, its renal interstitial fibrosis also has a certain inhibitory effect. The study showed that the RAAS inhibitor slowed the progression of the disease, the incidence of end-stage renal disease decreased significantly. This article will elaborate and analyze the progress of the application of RAAS inhibitors in the treatment of diabetic nephropathy to help more researchers enhance their understanding and improve the clinical efficacy of diabetic nephropathy.

Time-dependent Protein Expressions of MMP-2 and MMP-9 in Liver Contusion Rats after Impact / 法医学杂志

Objective To observe the protein expression patterns of matrix metalloproteinase (MMP)-2 and MMP-9 in the liver tissue of liver contusion rats at different time after impact. Methods Fifty healthy adult male SD rats were randomly and evenly divided into control group and experimental groups (1 h, 3 h, 6 h, 12 h, 18 h, 24 h, 3 d, 5 d, 7 d after liver contusion). A rat liver contusion model was established by a free-falling device. The rats were killed at corresponding time, and the contused hepatic lobes were extracted. The protein expressions of MMP-2 and MMP-9 in contused liver tissue of the rats in each group were observed by immunohistochemical staining (SP method) and Western blotting. Results After the liver contusion, the expression of positive cell and the protein semiquantitative result showed that the protein expression of MMP-2 enhanced at 6 h and peaked at 24 h, then decreased gradually at 3-5 d, and returned to normal levels at 7 d. The difference of expression between group and its previous adjacent group after 6 h (except 18 h) had statistical significance (P<0.05). The protein expression of MMP-9 rose obviously at 1 h after liver contusion and peaked at 18 h, then decreased gradually at 3-7 d which still higher than control group. The expression difference between group and its previous adjacent group (except 12 h and 24 h) had statistical significance (P<0.05). Conclusion The protein expressions of MMP-2 and MMP-9 in contused liver tissue after impact show good time-dependent patterns, which may provide important reference indicators for the time estimation of liver contusion.

Simultaneous determination of loganin, chiratin, secoxyloganin, rosmarinic acid and calceolarioside B in Shuangxiangpaishi granules by dual-wavelength HPLC method / 国际药学研究杂志

Objective To establish an HPLC method for simultaneous determination of loganin, chiratin, secoxyloganin, rosmarinic acid and calceolarioside B in Shuangxiangpaishi granules. Method HPLC colunmn was Alltima C18 column (4.6 mm×250 mm, 5 μm); mobile phase consisted of acetonitrile-methanol (1: 3) (A) -0.1% phosphate acid solution (B) with gradient elution; the column temperature was 35°C; the flow rate was 1.0 ml/min; all the injection volume was 20 μl; loganin, chiratin and secoxyloganin were detected at 240 nm, rosmarinic acid and calceolarioside B were detected at 330nm. Results The above mentioned five main ingredients had linearity in the given concentration range at 5.91-118.20 μg/ml (r=0.9995), 4.10-82.00 μg/ml (r=0.9991), 8.13-162.60 μg/ml (r=0.9993), 12.57-251.40 μg/ml (r=0.9998), 4.95-99.00μg/ml (r=0.9997), respectively. The average recoveries (n=6) and RSD were 96.88 (1.31), 99.09 (1.47%), 98.29 (1.59), 98.82 (1.42) and 97.51 (0.86), respectively. Conclusion This dual-wavelength HPLC method could simultaneously determine the contents of the five ingredients in Shuangxiangpaishi granules and can be used to evaluate their quality. The method is simple, accurate, sensitive and repeatable. It could be used as an efficient strategy for systematic quality evaluation of Shuangxiangpaishi granules.