This study is aimed to establish a high-performance liquid chromatography (HPLC) method for simultaneous determination of skimmin, scopolin and umbelliferone in Saussurea hieracioides. Samples were analyzed on a Wondasil C18-WR column (4.6 mm x 250 mm, 5 microm) with methanol (A) and water containing 0.1% phosphate (B) as mobile phases for gradient elution at a flow rate of 1.0 mL x min(-1). The detection wavelength and column temperature were set at 325 nm and 35 degrees C, respectively, and the sample size was 10 microL. The results showed that skimmin, scopolin and umbelliferone were simultaneously achieved within 40 min under the above conditions. A good linearity was observed in the range of 0.18-5.6 microg (r = 1.000 0), 0.060-1.8 microg (r = 0.999 9), 0.032-0.97 microg (r = 0.999 8) for skimmin, scopolin and umbelliferone, respectively, with the average recoveries of 99.16% (RSD = 0.41%), 100.3% (RSD = 0.79%), 102.2% (RSD = 0.87%). The method is simple, accurate and reproducible and can be used for the quality control of S. hieracioides.
Chromatography, High Pressure Liquid ; Coumarins ; analysis ; Glucosides ; analysis ; Medicine, Tibetan Traditional ; Reproducibility of Results ; Saussurea ; chemistry ; Umbelliferones ; analysis

OBJECTIVETo observe the effect of sequential method of nourishing yin and tonifying yang on serum sex hormone levels in rats with polycystic ovary syndrome (PCOS), and to explore its possible mechanisms.

METHODSPCOS rat models were established by injecting dehydroepiandrosteron (DHEA) subcutaneously. A total of 70 21-day female SD rats were randomly divided into seven groups, ten in each group: the normal control, the model control, the yin nourishing, the yang tonifying, the sequential method, the Westem medicine, and the combined medication. Rats in all groups were treated by gastrogavage for twelve successive days. Serum testosterone (T) and estradiol (E2) were detected by chemoluminescence method. Serum androstenedione (A), follicle stimulating hormone (FSH), luteinizing hormone (LH), insulin (INS), and insulin-like growth factor-I (IGF-I) were detected by radioimmunoassay (RIA).

RESULTSSerum levels of A, T, and E2 were significantly higher in the model control group than in the normal control group (P < 0.05, P < 0.01). Serum levels of A and T were obviously lower in the sequential group than in the model control group with significant difference (P < 0.01). The serum E2 level was obviously lower in the sequential group than in the model control group (P < 0.05). Serum levels of INS and IGF-I were significantly higher in the model control group than in the normal control group (P < 0.05). Serum levels of INS and IGF-I were significantly lower in the sequential group than in the model control group (P < 0.01).

CONCLUSIONSThe sequential method of nourishing yin and tonifying yang can lower serum levels of A, T, E2, INS, and IGF-I, and reverse its endocrine disturbance in PCOS rats. One of its mechanisms in decreasing serum levels of A, T, E2 might be possibly through lowering serum levels of INS and IGF-I.

Animals ; Female ; Gonadal Steroid Hormones ; blood ; Medicine, Chinese Traditional ; methods ; Polycystic Ovary Syndrome ; blood ; therapy ; Rats ; Rats, Sprague-Dawley

AIM: To evaluate the effects of the extract of Buddleja officinalis eye drops in basic tears secretory volume, tear film stability, expression of androgen receptors(AR) in castrated rats with dry eye, and to investigate the therapeutic effects of extract of Buddleja officinalis on dry eye caused by gonadal hormones level imbalance. METHODS:A total of 45 Wistar masculinity rats were divided at random into 9 groups, including normal group(A1,A2 and A3), model group(B1,B2 and B3), therapy group with extract of Buddleja officinalis eye drops(C1,C2 and C3). The "1" stood for being fed for 1 month, and "2" for 2 months, and "3" for 3 months. The dry eye model was established with orchiectomy on group B,C. Group C was treated with Buddleja officinalis extract eye drops for one month. All rats were checked with Schirmer Ⅰ test (SⅠt) and tear film break-up time (BUT). Expression of AR was analyzed by flow cytometer(FCM). RESULTS:The SⅠt value of group C was significantly higher than that of group B (P<0.01) and the BUT value of group C was significantly longer than that of group B (P<0.01), which indicated the eye drop could significantly keep basic tears secretory volume and tear film stability. And the expression of AR of group C was much higher than that of group B,which showed that available composition of the eye drops maybe display androgen-like activity.CONCLUSION:The main components of extract of Buddleja officinalis is the flavonoids which could significantly inhibit happening of dry eye of rat after androgen level lowered. Its mechanism is like androgen's and it could display androgen-like activity to keep basic tears secretory volume and tear film stability.

Aim To investigate the effects of berberine combined with bone marrow mesenchymal stem cells ( BM-MSCs) on the energy metabolism of human um-bilical vein endothelial cells ( HUVECs ) under the condition of high glucose. Methods ①The state of cell reproduction and cell proliferating activity were de-termined by MTT assay. ②The cell cycle was detected by flow cytometry ( FCM ) . ③DNA damage of cells was measured by comet tail assay. ④The contents of ATP, ADP and AMP were determined by high perform-ance liquid chromatography ( HPLC ) and the level of energy charge ( EC) was calculated. ⑤The expression of CCR and COX mRNA was detected by reverse tran-scription-polymerase chain reaction ( RT-PCR) . ⑥The expressions of COX and CCR were detected by Western blot. Results ①The proliferating activity of HUVECs declined apparently and the proliferation decreased af-ter high glucose intervention. Meanwhile, the quantity of cells during S +G2 dropped dramatically and there was certain degree of damage to DNA. The berberine and BM-MSCs respectively improved the proliferating activity and the proliferation in different degrees, in-creased the quantity of cells during S+G2 and promo-ted the repair of DNA ( P <0 . 01 ) , and so did the combination of the two, with a better effect than each of them alone. ②After high glucose intervention and the damage caused, the content of both ATP and ADP of HUVECs was reduced, and EC level also declined significantly, while the content of AMP increased. The berberine and BM-MSCs respectively up-regulated the content of ATP and ADP ( P<0. 01 ) , and so did the combination of the two, with a better effect than each of them alone. ③After high glucose intervention and the damage caused, the expression of COX, CCR mR-NA and protein decreased obviously. Yet, all of the three gained a dramatic increase when the berberine, BM-MSCs or the combination of the two were added ( P<0. 01 or P <0. 05 ) , among which the combination worked more effectively. Conclusions The berber-ine, BM-MSCs and the joint use of the two could im-prove the energy metabolism of HUVECs, which had been damaged by high glucose, probably because the berberine and BM-MSCs could up-regulate the expres-sion of COX, CCR mRNA and protein, which leads to the hydrolyzation of glucose oxide and thus the im-provement of blood environment and the enhancement of glucose's supply and intake of HUVECs. Then, here comes the final result: the improvement of the energy metabolism of damaged vascular endothelial cells by high glucose.

OBJECTIVE: To study the effect of low-dose dopamine adjuvant therapy on inflammatory factors and prognosis in preterm infants with necrotizing enterocolitis (NEC). METHODS: A total of 100 preterm infants with NEC from June 2017 to June 2019 were enrolled and divided into a dopamine treatment group and a conventional treatment group using a random number table, with 50 infants in each group. The infants in the conventional treatment group were given symptomatic treatment, and those in the dopamine treatment group were given low-dose dopamine adjuvant therapy in addition to the conventional treatment. ELISA was used to measure the levels of C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), and interleukin-8 (IL-8). The two groups were compared in terms of time to relief of clinical symptoms, fasting time, treatment outcome, prognosis, and adverse reactions. RESULTS: Both groups had significant reductions in the levels of CRP, TNF-α, and IL-8 after treatment, and the dopamine treatment group had significantly lower levels of these markers than the conventional treatment group after treatment (P<0.05). Compared with the conventional treatment group, the dopamine treatment group had significantly shorter time to defecation improvement, time to relief of abdominal distension and diarrhea, and fasting time (P<0.05), a significantly higher response rate (P<0.05), and a significantly lower surgery rate (P<0.05). There were no significant differences in the mortality rate and incidence of adverse events between the two groups (P>0.05). CONCLUSIONS Low-dose dopamine adjuvant therapy can effectively improve the levels of inflammatory factors and clinical symptoms in preterm infants with NEC and has good safety, and therefore, it holds promise for clinical application.

Aged ; Aged, 80 and over ; Delivery of Health Care ; Family Characteristics ; Female ; Home Care Services ; Humans ; Male ; Middle Aged ; Rural Population ; Surveys and Questionnaires

This study aimed to explore the impact of depression caused by chronic unpredictable mild stress (CUMS) on in vivo activity of six kinds of CYP450 isoforms in rats. According to 'Katz' method, the model of CUMS was established. Tolbutamide, chlorzoxazone, theophylline, midazolam, omeprazole and dextromethorphan were chosen as probe substrates of CYP2C6, CYP2E1, CYP1A2, CYP3A2, CYP2D1 and CYP2D2 of rats. Plasma concentration of six kinds of CYP450 in control group and model group were determined by LC-MS/MS and computed pharmacokinetic parameters. Consequently, metabolism of theophylline and chlorzoxazone accelerated significantly (P < 0.01), but tolbutamide, dextromethorphan, omeprazole and midazolam had no significant difference. The present study proved that depression caused by CUMS had strong induction to CYP1A2 and medium induction to CYP2E1.
Animals ; Chlorzoxazone ; metabolism ; Chromatography, Liquid ; Cytochrome P-450 Enzyme System ; metabolism ; Depression ; Dextromethorphan ; metabolism ; Liver ; enzymology ; Midazolam ; metabolism ; Omeprazole ; metabolism ; Rats ; Stress, Physiological ; Tandem Mass Spectrometry ; Theophylline ; metabolism ; Tolbutamide ; metabolism

Objective To establish an isolated rat pancreas perfusion technique,a method for the precise measurement of insulin secretion in vitro.Methods An isolated rat pancreas perfusion technique was applied in the study of insulin secretion from?-cells in 10 high-fat diet-induced obese Wistar rats.Results For the assessment of the functional integrity of the perfused pancreas,the isolated pancreas of 6 rats met all the criteria: (1)The constancy of perfusion pressure was kept over the whole experiment time[(70?5)mm Hg,1 mm Hg= 0.133 kPa].(2)The duodenal peristaltic activity of isolated pancreas and duodenum block was present after perfusion experiment.(3)Total insulin response to arginine stimulation was significantly increased as compared with glucose stimulation[maximum insulin secretion rate:(987?100)?U/min vs(545?50)?U/min,P

Objective:To study the value of small dose sufentanil in anesthesia in caesarean section for prevention of chill and dragging pain. Methods:A total of 84 cases of caesarean section maternal in our hospital were choosen for study as the object. And they were randomly divided into two groups, with 42 cases in each group. The experimental group implements the sufentanil 5μg, control group was given 0.5%ropivacaine. Chills,dragging pain and other adverse reactions were compared. Results:The incidence rate of level 0 chills in experimental group was 80.95%,and it's higher than that 50.00%of control group (P<0.01) . Maternal pull pain incidence rate of experiment group was 14.92%, while the control group was 73.81%(P<0.01) . The other adverse reaction rate in experimental group was lower than that in the control group (P<0.01) . Conclusion:Small doses sufentanil anesthesia for cesarean section can significantly reduce the occurrence of adverse reactions such as pain and chills,safe and effective,it's worthy of clinical popularization and application.

OBJECTIVETo study the gap junction mechanisms for synergistic effects of liuwei di-huang pill (LDP) containing serum on HSV-tk/GCV suicide gene therapy of mouse malignant melanoma B16 cells.

METHODSThe LDP containing serum (2.5% LDP serum group, 5.0% LDP serum group, and 10.0% LDP serum group) and the control serum group were set up. The effects of LDP on mRNA expressions of Cx26 and Cx43 in mouse malignant melanoma B16 cells were detected by RT-PCR assay. The effects of LDP on protein expressions of Cx26 and Cx43 in B16 cells were detected using Western blot and indirect immunofluorescence assay. The interference efficiencies of Cx26-309, Cx26-337, Cx26-367, Cx26-2098 SiRNA on Cx26 gene in B16 cells were detected using RT-RCR technique. Cx26-2098 SiRNA, due to the optimal interference efficiency, was chosen to interfere Cx26 gene, and the bystander effect of LDP + HSV-tk/GCV was observed. The effects of the gap junctional intercellular communication (GJIC) inhibitor glycyrrhetinic acid on the killing of LDP + HSV-tk/GCV system to B16 cells were detected by MTT assay. In this experiment, the control serum group, 2.5% LDP serum group, 5. 0% LDP serum group, 10.0% LDP serum group, the control serum combined GCV group, 2.5% LDP serum combined GCV group, 5.0% LDP serum combined GCV group, 10.0% LDP serum combined GCV group, 2.5% LDP serum combined GCV + glycyrrhetinic acid group, 5.0% LDP serum combined GCV + glycyrrhetinic acid group, 10.0% LDP serum combined GCV + glycyrrhetinic acid group were set up. The final concentration of GCV was 20 micromol/L. The final concentration of glycyrrhetinic acid was 50 micromol/L.

RESULTSLDP containing serum could increase the protein and mRNA expressions of Cx43 in B16 cell in a dose-dependent manner. It had bi-directional regulation on the Cx26 protein and mRNA expressions. The low dose LDP had inhibition while high dose LDP could up-regulate its expression. After SiRNA interfered Cx26 gene, there was no obvious change in the bystander effect of LDP combined suicide gene therapy between before and after interference. There was significant reduction in the inhibition rate between before (48.75%, 59.39%, and 69.28%) and after blockage (29.14%, 38.71%, and 58.13%) of glycyrrhetinic acid in the 2. 5%, 5.0%, 10.0% LDP serum combined GCV groups (P <0.01).

CONCLUSIONThe synergistic effects of LDP containing serum on HSV-tk/GCV suicide gene therapy in mouse malignant melanoma B16 cells were correlated with the gap junction mechanisms, which might be achieved through increasing the expressions of Cx26 and Cx43.

Animals ; Cell Line, Tumor ; Connexin 26 ; Connexin 43 ; metabolism ; Connexins ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Gap Junctions ; metabolism ; Genetic Therapy ; Male ; Melanoma ; therapy ; Mice ; Rats, Sprague-Dawley ; Serum