OBJECTIVE To map a comprehensive metabolic pathway of herbacetin in rats, specifically, to elucidate the biotransformation of herbacetin in vivo and to simultaneously monitor the pharmacokinetic process of both parent drug and its major metabolites. METHODS liquid chromatography/ion trap mass spectrometry (LC/MSn) and ultra-liquid chromatography coupled with mass spectrometry (UPLC/MS) were combined in the current study for qualitative and quantitative determinations of herbacetin and its metabolites in bile, urine and feces after both oral and intravenous administration of herbacetin to rats. Enzyme kinetic studies on the intestinal and hepatic metabolism of herbacetin were further conducted to elucidate metabolic profiles of herbacetin in rat tissues and organs. Additionally, plasma concentration profiles of herbacetin and its metabolites in rats were obtained to characterize the overall pharmacokinetic behavior of herbacetin. RESULTS It was found that herbacetin was excreted primarily from rat urine in the form of glucuronide-conjugations. Subsequent in vitro enzyme kinetic studies and in vivo pharmacokinetic investigations suggested an extensive hepatic metabolism of herbacetin and the high exposure of herbacetin- glucuronides in systemic circulation. The clearance, half- life and bioavailability of herbacetin in rats were determined as (16.4±1.92)mL·kg-1·min-1, (11.9±2.7)min, and 1.32%, respectively. On basis of these findings, a comprehensive metabolic pathway of herbacetin in rats was composed. In addition, a physiology based pharmacokinetic (PBPK) model was successfully developed with the aid of the GastroPlus to simulate the pharmacokinetic process of herbacetin in rats. Application of the PBPK modeling can provide a useful starting point to understand and extrapolate pharmacokinetic parameters among different species, populations, and disease states. CONCLUSION After oral administration, herbacetin was subjected to colonic degradation and extensive first pass metabolism, with glucuronidation as its dominating in vivo metabolic pathway.

OBJECTIVE To investigate the effect of hypoxia on the pharmacokinetic process of salidrosidein rats and to explore its underlying mechanisms. METHODS The Caco-2 cell monolayerwas exposed to 1% oxygen (O2) concentration for 24 h to build the hypoxiccell model. The transportation mode of salidroside was investigated with the aid of this hypoxia model by detecting the apparent permeability coefficient(Papp). Healthy Sprague Dawley (SD) rats were exposed to 9% O2 for 72 h for the construction of hypoxic rat model. Liver sample was subsequently collected from the hypoxic rats with an aim to identify enzymes responsible for salidroside metabolism. The expression levels of sali?droside-transporting and salidroside-metabolizing enzymes, including Sodium-dependent glucose cotrans?porters (SGLT1), β-glucosidase (GBA3)and sulfotransferase (SULT2A1), were thereafter detected by RT-PCR and Western blot. The metabolic activity of GBA3 and SULT2A1 was monitored by rat liver microsome incubation.In addition, the renal function of rats under hypoxia was assessed by detecting concentrations of blood urea nitrogen and creatinine. RESULTS The AUC and t1/2 values of salidroside in hypoxic rats were more than doubled, while the in vivo clearance was significantly reduced. Mechanistic study demonstrated that the PappA- B/PappB- A eualsto 10.3, indicating the potential active transport of salidrosile. The expression of SGLT1 and GBA3 was significantly decreased, which indicated a reduced metabolism of salidroside under hypoxia. Moreover, rat under hypoxia was found to suffer from renal dysfunction, with an abnormal value of blood urea nitrogen. CONCLUSION Due to the reduced metabolism and the abnormal renal function under hypoxia, the systemic exposure of salidroside in rats was signifi?cantly enhanced.

Objective To quantifiably measure the methylation frequency of 18 CpG sites in the 3′region of L1 gene and long control region(LCR) gene of HPVl6 DNA,and study the relationship between HPVl6 DNA methylation and severity of cervical lesions. Methods A total of 10 cases Normal/low?grade squamous intraepithelial lesion(Normal/LSIL),10 cases of high?grade squamous intraepithelial lesion(HSIL),and 10 cases of cervical cancer(CC)were recruited for the study. The relationship between severity of cervical lesions and HPV16 DNA methylation was analyzed by bisultlte?pyrosequencing. Results The methylation rate was highest in Normal/LSIL at position 7 089 located in 3′?L1,followed by CC. The low?est was found in HSIL. The difference in methylation percentage among the three lesions was significant(P=0.006). In 7 134,the proportion meth?ylation was also different among three groups(P=0.01),difference in methylation percentage between Normal/LSIL and CC,as well as Normal/LSIL and HSIL was significant(P=0.038,0.017). Conclusion The methylation status of CpG sites 7 089 and 7 134 in the 3′region of L1gene is asso?ciated with the severity of cervical disease. The quantification of HPV DNA methylation can be used for cervical disease screening in clinical samples.

Objective To observe the clinical efficacy of successive trigger needling plus motorial needling in treating external humeral epicondylitis.Method Sixty-two eligible subjects with external humeral epicondylitis were randomized into a treatment group and a control group,31 cases each.The treatment group was intervened by successive trigger needling plus motorial needling,and the control group was treated with ordinary acupuncture.The two groups were both treated once a day.After 10 sessions of treatment,the changes of Visual Analogue Scale (VAS) and Activities of Daily Living (ADL) scores were observed,and the clinical efficacies were compared between the two groups.Result The VAS and ADL scores were changed significantly respectively after 1-week and 2-week treatment in both groups (P＜0.05).After the treatment,the changes of VAS and ADL scores in the treatment group were significantly different from those in the control group (P＜0.05).The total effective rate was 96.8％ in the treatment group versus 93.5％ in the control group,and the difference was statistically significant (P＜0.05).Conclusion Successive trigger needling plus motorial needling is an effective approach in treating external humeral epicondylitis,and it can improve the ADL.

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a common condition obsessing urologists and patients. It is also known as a heterogeneous syndrome, with varied etiologies, progression courses and responses to treatment. Based on the deeper insights into its pathogenesis and re-evaluation of its clinical trials, a novel phenotypic classification system UPOINT has been developed, which clinically classifies CP/CPPS patients into six domains: urinary (U), psychosocial (P), organ-specific (O), infection (I) , neurologic/systemic (N) and tenderness of pelvic floor skeletal muscles (T), and directs individualized and multimodal therapeutic approaches to CP/CPPS. This review systematically summarizes the theoretical foundation, clinical characteristics of UPOINT and treatment strategies based on the UPOINT phenotypic classification system.
Chronic Disease ; Humans ; Male ; Pelvic Pain ; classification ; diagnosis ; therapy ; Phenotype ; Prostatitis ; classification ; diagnosis ; therapy ; Severity of Illness Index

Objective: To investigate the distribution of human papillomavirus (HPV) infection characteristics and genotypes in Shenyang area of Liaoning province. Methods: HPV genes were detected in cervical exfoliated cells from 55, 548 patients by amplification and diversion hybridization. Results: A total of 9, 566 patients were positive for HPV infection with a positive rate of 17.22%. Additionally, the positive rate of high risk HPV infection was 14.57% and the positive rate of single genotype HPV infection was 13.63%. Totally, 12, 360 HPV viruses were detected. Among them, 10, 879 HPV viruses were classified into high risk genotypes (10, 879 out of 12, 360, 88.02%). The genotypes in women with ages less than 20 were 16/11/6/51/58/52 genotypes; the susceptible HPV genotypes in other women were 16/58/52/53/39/51/81 genotypes. Conclusions HPV infections in Shenyang are mainly infections with high risk viruses and single infection. The infection rate and genotype distribution of HPV are different in different age groups. More suitable HPV vaccine prophylaxis can be taken according to the epidemic characteristics of HPV in this area.

Objective:To explore the features of atherosclerotic plaque in patients with type 2 diabetes mellitus by contrast-enhanced ultrasound (CEUS), and to analyze the correlation between the features and leukocyte count in peripheral blood.Methods:A total of 30 type 2 diabetic patients with carotid atherosclerotic plaque treated in Tangdu Hospital of Air Force Military Medical University who underwent CEUS from March 2018 to September 2021 were included as the diabetes group. Thirty-two non-diabetic patients with carotid atherosclerotic plaque who were matched for age and sex during the same period served as the control group. The optimized plaques were performed by CEUS to obtain parameters including peak (P), time to peak (TP), sharpness (S) and area under the curve (AUC) of time-intensity gamma fitting curve (FC curve). The differential count of leukocytes in peripheral blood was detected by the automatic hematology analyzer. Spearman rank correlation and multiple linear regression were used to analyze the correlation between CEUS parameters of plaque and the differential leukocyte count in peripheral blood.Results:①The peripheral blood leukocyte count, monocyte count and monocyte proportion in the diabetes group were higher than those in the control group (all P<0.05). ②P and AUC in the diabetes group were higher than those in the control group (all P<0.05). ③There was linear relationship( B=75.440, P<0.001) and positive correlation ( rs=0.929) between P and peripheral blood monocyte count in the diabetes group. Conclusions:The neovascularization density and perfusion in carotid plaques in patients with type 2 diabetes mellitus are increased, which may be related to the increase of leukocytes in circulation, especially monocytes.

Objective To explore the prevalence and analyze the related risk factors for post-stroke depression among middle-aged and elderly stroke patients of Mongolia and Han nationality in Inner Mongolia Autonomous Region.Methods A stratified multistage random cluster sampling method was used to investigate the prevalence of stroke in the population aged ≥45 years in different areas of Inner Mongolia Autonomous Region.A total of 11 088 people were investigated and 498 were stroke patients among them.A set of scale assessment and the general situation were used to investigate stroke patients and 443 patients completed the whole survey.Results The total prevalence of post-stroke depression (PSD) was 41.08％ in 443 stroke patients,among which the prevalence rate was 40.57％ for Mongolian population and 41.85％ for Han population,and no significant difference was found between Mongolian and Han nationality(x2=1.372,P=0.504).There were significant differences in the types of stroke between the Mongolian and Han nationality (x2 =7.347,P=0.025).The age (t=4.321,P=0.000),educational level (x2 =27.036,P=0.001) and economic burden (x2=27.877,P=0.000) were statistically significant between Mongolian and Han nationality.The differences of frequency of stroke (x2 =6.545,P=0.011),economic burden (x2 =16.148,P=0.001),cognitive dysfunction (x2 =9.065,P=0.003),daily living ability (x2 =34.466,P =0.000),alcohol consumption history(x2=4.516,P=0.034)were statistically significant.Logistic regression analysis showed that alcohol consumption history,economic burden,the frequency of stroke,and cognitive dysfunction were the influencing factors of PSD.Conclusion PSD is one of the important factors affecting the post-stroke psychological burden in Inner Mongolia Autonomous Region.There is no national difference in the prevalence of PSD among Mongolian and Han people,which provides a theoretical basis for the treatment and intervention of PSD.

OBJECTIVETo investigate the hemostatic effect of spleen-invigorating, qi-replenishing and blood-containing formula on simvastatin-induced zebrafish hemorrhage model, and to compare with the effect of clearing heat and cooling blood formula.

METHODSZebrafishes from breed A B line were treated with 0.5 µmol/L simvastatin for 24 hours to establish zebrafishes hemorrhage model. Under strict blinded experimental conditions, the above mentioned zebrafishes were then treated with experimental drug of different concentrations at the maximum non-lethal dose. The intervention effect of spleen-invigorating, qi-replenishing and blood-containing formula was comprehensively assessed by examining the main observational parameters, such as bleeding reduction rate and hemostasis rate while referring to additional parameters, such as blood flow, improvement rate of blood flow, velocity of movement, improvement rate of motion, which are characteristics of spleen qi deficiency.

RESULTSWhen the hemostatic effect of experimental drug B1 at the concentrations of 500 and 1 000 µg/ml, zebrafish bleeding rates were 30% and 15%, the hemostatic rate was 60% and 80%, respectively; when the experimental drug B2 at concentration of 500 and 1 000 µg/ml, Zebrafish bleeding rates were 45% and 40%, the hemostatic rate was 40% and 47%, respectively, showing that experimental drug B1 was superior to B2 in terms of decreasing bleeding rate and improving hemostatic effect in zebrafish. In the equal concentration, the experimental drug B1 was superior to B2 in terms of increasing and improving the blood flow of hemorrhagic zebrafish. Promotion and improvement of motion: in equal concentration, experimental drug B1 was superior to B2 in terms of promoting the motion velocity and increasing the improving rate of motion in zebrafish.

CONCLUSIONThe spleen-invigorating, qi-replenishing and blood-containing formula displays a good hemostatic effect on simvastatin-induced hemorrhage of zebrafish. It also boosts the blood flow and motion velocity in hemorrhagic zebrafish, therefore, providing an experimental basis for the treatment of syndrome of spleen failing to control blood by spleen-invigorating, qi-replenishing and blood-containing formula.

OBJECTIVEMucopolysaccharidosis type II (MPS II, Hunter syndrome, OMIM 309900) is an X-linked recessive lysosomal storage disease resulting from a deficiency of iduronte-2-sulphate sulphatase (IDS). The present study aimed to establish an enzyme assay method for IDS activity for carrying out postnatal and prenatal diagnosis of MPS II by means of IDS activity assay on plasma, uncultured chorionic villi (CV) and cultured amniotic fluid cells (AF cell) using a new synthesized substrate.

METHODSA fluorigenic substrate (4-methylumbelliferyl-alpha-iduronate-2-sulphate, MU-alpha-Idu-2S) was used for the assay of IDS activity. IDS activity in plasma was determined for diagnosis of the proband. Prenatal diagnosis in 10 pregnancies at risk was carried out according to IDS activity on uncultured CV at 11th week or on cultured AF cell at 18th week of gestation. At the same time, IDS activity was also determined in the maternal plasmas to observe the change of IDS activity in pregnancy. The fetal sex determination was performed by PCR amplification of the ZFX/ZFY genes.

RESULTThe IDS activity in plasma of normal controls and obligate heterozygotes were 240.2 - 668.2 nmol/(4 hxml) and 88.7 - 547.9 nmol/(4 hxml), respectively, while the enzyme activities in plasmas were in the range of 0.3 - 18.6 nmol/(4 hxml) in affected male. The IDS activities were 37.2 - 54.9 nmol/(4 hxmg protein) and 21.4 - 74.4 nmol/(4 hxmg protein) in CV and cultured AF cells respectively. Out of 50 suspected cases, 46 were diagnosed as having MPS II and 4 were excluded. Prenatal diagnosis was performed on 10 pregnancies at risk. Four of 5 male fetuses [IDS activity were 4.7, 1.8, 7.0 nmol/(4hxmg protein) in CV, 0.6 nmol/(4 hxmg protein) in AF cell] were diagnosed as having MPS II and the other 5 fetuses were normal females [IDS activity were: 48.7, 5.9, 25.2 nmol/(4 hxmg protein) in CV, 55.2, 40.9 nmol/(4 hxmg protein) in AF cell]. Increased IDS activity was observed in plasma of the pregnant women with unaffected fetuses, while the IDS activity decreased in pregnancies with affected fetuses. IDS activity of one female fetus was very low [5.9 nmol/(4 hxmg protein)], but the IDS activity in maternal plasmas increased, this fetus was a normal female.

CONCLUSIONSThe method using a synthesized fluorigenic 4-methylumbelliferyl-substrate was a sensitive, rapid and convenient assay of IDS activity and was reliable for early prenatal diagnosis. Determination of fetal sex would be helpful in excluding the female fetus with low IDS activity from being considered as an affected male fetus. It would be further helpful if IDS activity in maternal plasma was taken into account.

Amniotic Fluid ; cytology ; enzymology ; Cells, Cultured ; Child ; Child, Preschool ; China ; epidemiology ; Chorionic Villi ; enzymology ; Chorionic Villi Sampling ; Enzyme Assays ; methods ; Female ; Fetus ; enzymology ; Fluorometry ; methods ; Heterozygote ; Humans ; Hymecromone ; analogs & derivatives ; Iduronate Sulfatase ; blood ; metabolism ; Iduronic Acid ; analogs & derivatives ; Karyotyping ; Male ; Mucopolysaccharidosis II ; diagnosis ; enzymology ; epidemiology ; Polymerase Chain Reaction ; Pregnancy ; Pregnancy, High-Risk ; blood ; Prenatal Diagnosis ; methods ; Reference Values ; Sex Factors