Objective: To establish a method to determine 4 active ingredients in Jindan tablets by HPLC.
Methods: Agilent Eclipse X DB C₁₈ (4.6 mm×250 mm, 5 μm) column was adoped using acetonitrile (phase A) -0.1% phophoric acid solution (phase B) as the mobile phase with gradient program at the flow rate of 1.0 mL·min^-1, the detection wavelength was 270 nm, Column temperature was set at 30 ℃.
Results: The linear ranges of gentiopicrin, polydatin, quercetin and emodin were 7.875-78.75 μg·mL^-1（r=0.999 9）, 6.75-67.50 μg·mL^-1（r=0.999 7）, 7.726-77.26 μg·mL^-1（r=0.999 4）, 3.809-38.09 μg·mL^-1（r=0.999 8), respectively, the peak area had a good linear relationship with the mass concentration of 4 components, the average recovery were 99.31%, 99.21%, 99.04%, 99.59%, (n=6) respectively, and RSDs were 1.86%, 1.24%, 1.37%, 1.15%, respectively.
Conclusion: This method is reliable, repeatable and stable, and has strong specificity. This method can provide a reference for the quality control and standard improvement of Jindan tablets.

OBJECTIVETo explore the clinical efficacy and action mechanism of acupuncture for premature ovarian failure (POF).

METHODSAccording to prospective cohort study design, 30 cases were included. Based on theory of regulating Chong and Conception Vessels as well as soothing the liver and calming the nerves, acupuncture at Guanyuan (CV 4), Dahe (KI 12), Taixi (KI 3), Taichong (LR 3), Baihui (GV 20) was applied, three times per week and 3 months were considered as one session. Totally two sessions were performed. The menstruation condition, estradiol (E2), follicle-stimulating hormone (FSH) and scores of the clinical perimenopausal symptoms were taken as treatment outcomes.

RESULTSThe total effective rate was 86.7% (26/30) and the cured rate was 16.7% (5/30). The recovery rate of menstruation was 16.7% (5/30) and the regain rate of menstruation was 56.7% (17/30). After the treatment, the level of E2 was significantly increased from (45.41 +/- 18. 09) pmol/L to (59.07 +/- 24.21) pmol/L (P < 0.05), and the score of perimenopausal symptoms was obviously reduced from 14.28 +/- 8. 30 to 4.04 +/- 3.28 (P < 0.01). However, no statistical differences could be seen in FSH before and after treatment (P > 0.05).

CONCLUSIONAcupuncture has certain effect on improving menstruation and perimenopausal symptoms in POF patients, which is likely to he related with increasing the level of E2.

Acupuncture Therapy ; Adolescent ; Estradiol ; metabolism ; Female ; Humans ; Luteinizing Hormone ; metabolism ; Primary Ovarian Insufficiency ; metabolism ; therapy ; Prospective Studies ; Young Adult

Objective To study interleukin-1 receptor(IL-1R) and connexin(Cx36) gene expression following Mg 2+-free-induced seizures in cultured developing neuron. Methods Rat embryo cortical neurons cultured for 6 and 17 days were exposed to Mg 2+-free media to induce seizure. At different time after Mg 2+-free treatment, real-time RT-PCR was used to detect IL-1R and Cx36 mRNA expression. Results 1. IL-1R mRNA expression transiently decreased after Mg 2+-free treatment in neurons cultured for 6 and 17 days in vitro. Then the levels of IL-1R mRNA expression recovered in neurons cultured for 6 days, but IL-1R mRNA expression were increased in neurons cultured for 17 days compared with control group and the peak was at 24 hours. 2. In neurons cultured for 6 days in vitro, Cx36 mRNA expression increased after Mg 2+-free treatment compared with control group, the peak was at 24 hours. But in neurons cultured for 17 days in vitro, Cx36 mRNA expression decreased at 6 hours after Mg 2+-free treatment compared with control group, the peak was at 24 hours. Conclusions IL-1R mRNA and Cx36 mRNA expression following Mg 2+-free-induced seizures are different between the neurons cultured for 6 and 17 days in vitro. This is possibly related to the different neuron injury between 6 and 17 days in vitro following seizures.

PEG-modified magnetic Fe3O4 (Fe3O4-PEG) nanoparticles were sythesized using a solvothermal reaction and characterized with transmission electron microscopy (TEM) and thermo gravimetric analysis (TGA). The photothermal effect and photothermal destruction of cancer cells were evaluated. Then the doxorubicin loaded Fe3O4-PEG (DOX-Fe3O4-PEG) nanoparticles were prepared. The cytotoxicity and combined chemotherapy/photothermal therapy (PTT) effect were investigated. Uniform PEG coated Fe3O4 nanoparticles with particle size of 155 nm were obtained in the experiment. The loading and release of doxorubicin on Fe3O4-PEG were pH-dependent. The drug loading capacity in water was 21%. The results of MTT indicated a good biocompatiblity of Fe3O4-PEG nanoparticles and high cytotoxicity of DOX-Fe3O4-PEG. In combined therapy experiment, photothermal therapy demonstrated unambiguously enhanced chemotherapy efficacy. In conclusion, the obtained Fe3O4-PEG nanoparticles which exhibit good photothermal effect and drug loading capacity can be used for chemotherapy and photothermal therapy. The synergetic anti-tumor activity indicates the potential for the combined application of chemotherapy and photothermal therapy in cancer treatment.
Antibiotics, Antineoplastic ; administration & dosage ; pharmacology ; Cell Survival ; drug effects ; Doxorubicin ; administration & dosage ; pharmacology ; Drug Carriers ; Ferrosoferric Oxide ; chemistry ; Humans ; Hyperthermia, Induced ; MCF-7 Cells ; Magnetite Nanoparticles ; chemistry ; Particle Size ; Polyethylene Glycols ; chemistry

Objective To investigate the clinical and electroencephalogram (EEG) characteristics of children with electrical status cpilepticus during sleep (ESES), and the response to medication therapy. Methods AEEG and VEEG including an entire sleeping c-ycle were performed on 26 patients with ESES. The clinical and EEG changes, neuropsychological impairment and the response to medication therapy were followed up. Results Twenty five patients had seizures,21 cases had normal psychomotor development before ESES. After the onset of the disease,Fifteen cases developed language disorder, 16 cases developed psychological and behavior abnormalities, 13 cases had both of the problems Seventeen patients belonged to epileptic syndrome. Patients in this cohort had good response to clonazepam and valproate treatment. Cortical steroid could dispel the electrical discharge. Eighteen patients had been followed up. Seizures stopped in 15 cases after treatment ESES disappeared in 16 cases, 4 of them still had neuropsychological impairment ESES sustained in 2 cases Conclusions ESES is a specific EEG phenomenon. Continue epileptic form discharge during non - rapid cye movement sleep is the major cause of neuropsychological impairment in patients with ESES. To control the seizures and electrical state are very important for the prevention and treatment of neuropsychological impairment.

Objective To study the expression level of thyroid insulin-like growth factor-Ⅰ (IGF-Ⅰ) in iodine deficiency and excess mice and the effect of thyroid gland IGF-Ⅰ in the thyroid morphological change. Methods Forty-eight Balb/c mice were chosen as studied objects,weighing about 16 g. They were divided into three groups: low iodine(LI,iodine content of 50 μg/kg in feed,drinking deironized water) group,normoi(NI,iodine content of 300 μg/kg in feed,drinking deironized water) group and high(HI,iodine content of 300 μg/kg in feed,iodoine of content 14 700 μg/kg in drinking) group,16 mice in each group. Mice were put to death after 12 weeks and taken out of their thyroid gland. The body weight,absolute and relative weights of thyroid gland were measured and the morphological change of thyroid gland were observed under microscope. The expression levels of thyroid gland IGF-Ⅰ mRNA and protein were detected by RT-PCR and immunohistochemistry,respectively. Results There were statistical significances between groups of thyroid absolute and relative weights(F = 315.881,405.921,all P < 0.01). LI group [(10.71±4.03) mg,(44.98±15.39)mg/100 g body weight]and HI group [(3.42±1.17)mg,(13.50± 3.89)mg/100 g body weight]had heavier thyroid absolute and relative weights than NI group[(2.11±0.53)mg,(8.35±1.98)mg/100 g body weight,all P < 0.01]. Under microscopy,the thyroid follicle capacity grew down and the follicle quantity grew up in LI group,the epithelium was stylolitic,the colloid diminished or absence in follicular cavity,while HI group presented colloid accumulation without follicular hyperplasia. The expression level of thyroid gland IGF-Ⅰ mRNA in LI group(1.03±0.32) was more than that in NI(0.65±0.19) and HI(0.59± 0.20) groups(F= 7.518,P< 0.01). In contrast to NI,there was no difference in the expression level of thyroid gland IGF-Ⅰ mRNA in HI group(P > 0.05). The brownish particles of LI group were more than NI and HI groups in the thyroid follicle epithelium by immunohistochemistry,while HI group was less than NI group. Conclusions The mice of iodine deficiency presented follicular hyperplasia goiter,the mice of iodine excess presented colloid accumulative goiter. The change of IGF-Ⅰ mRNA and protein expression may participate morphologleal change,indicating autocrine IGF-Ⅰ of thyroid gland may play an important role in regulating goiter formation.

The aim of this paper was to investigate the effect of terpene penetration enhancers on membrane fluidity and membrane potential using HaCaT keratinocytes, and study the potential mechanisms of these terpene compounds using as natural transdermal penetration enhancer. Six terpene compounds, namely menthol, limonene, 1,8-cineole, menthone, terpinen-4-ol and pulegone, were chosen in this study on account of their good penetration-enhancement activities. The cytotoxicity of these terpene compounds was measured using an MTT assay. The fluorescence recovery after photobleaching (FRAP) technique was employed to measure the change of membrane fluidity of HaCaT cells. The flow cytometer was used to study the alteration of membrane fluidity of HaCaT cells, and investigate the effect of terpene compounds on intracellular Ca2+. It was found that 6 terpene compounds possessed low cytotoxicity in comparison to the well-established and standard penetration enhancer azone. Those terpene compounds could significantly enhance HaCaT cells membrane fluidity and decrease HaCaT cells membrane potentials. Meanwhile, after treated with various terpene compounds, the Ca2(+)-ATPase activity and intracellular Ca2+ of HaCaT cells was decreased significantly. Terpene penetration enhancers perhaps changed the membrane fluidity and potentials of HaCaT cells by altering the Ca2+ balance of the cell inside and outside, resulting in the low skin permeability to increase the drug transdermal absorption.
Cell Line ; Drugs, Chinese Herbal ; pharmacokinetics ; Humans ; Keratinocytes ; drug effects ; metabolism ; Membrane Fluidity ; drug effects ; Skin Absorption ; drug effects ; Terpenes ; pharmacokinetics

OBJECTIVETo study cortical neuron injury following recurrent epileptiform discharges induced by magnesium-free treatment in vitro.

METHODSCultured embryo cortical neurons were exposed to magnesium-free media for 3 h, then they were returned to regular media containing normal level magnesium. At different time after Mg(2+)-free treatment, trypan blue staining and determination of LDH activity were used to determine the cell viability, flow cytometry was applied to measure neuronal apoptosis, and MTT assay to study metabolic rate.

RESULTS(1) Neuronal morphology on light microscopy following Mg(2+)-free treatment showed that there were no prominent alterations. (2) At different time (6, 12, 72 h) after Mg(2+)-free treatment, neuronal viability by trypan blue staining and LDH activity showed modest changes compared with time-matched control in different culture days (6, 12, 17 d) (P > 0.05). (3) Cell apoptosis increased mildly at different time after Mg(2+)-free treatment in neurons cultured for different days, but the increase was not significant (P > 0.05). (4) Metabolic rate decreased at 6 h after Mg(2+)-free treatment (P < 0.05) in neurons cultured for 6 d, and was 86.4% of that of the control; while the rate at 24 h in neurons cultured for 12 d and 17 d also decreased (P < 0.05), being 78.7% and 70.9%, respectively, of that of the control.

CONCLUSIONSThese findings demonstrated that the injury occurred on cultured cortical neurons caused by magnesium-free-treatment-induced recurrent epileptiform discharges was mainly functional and relatively mature neurons displayed more severe and much later mitochondrial function impairment than immature neurons.

Animals ; Cerebral Cortex ; embryology ; Culture Media ; chemistry ; pharmacology ; Culture Techniques ; Magnesium ; pharmacology ; Neurons ; drug effects ; pathology ; Rats ; Rats, Wistar ; Seizures ; physiopathology

Abstract: Objective To analyze the effect of nutrition package on the nutritional status and prevalence of children in rural areas of Hainan Province, and provide scientific basis and suggestions for further improving the nutritional and health status of children in this region. Methods　Four cities and counties were randomly selected as the intervention group, and four cities and counties matched with the intervention group in terms of population, economy, social culture, maternal and child health work foundation of township health centers, physical nutrition and health status were selected as the control group.With the combination of monitoring and prospective cohort study, infants in the intervention group and the control group were studied from June 1, 2020, and they were intervened for 12 months with supplementary food nutrition package. Before and after intervention, the nutrition and health status of infants aged 6-24 months in the intervention group and the control group were investigated to evaluate the nutritional and health effects of supplementary food nutrition package for infants aged 6-24 months in rural Hainan Province. Results　A total of 999 infants were investigated, including 427 in the intervention group and 572 in the control group. After 12 months of nutritional intervention, there was no significant difference in weight-for-age Z-score (WAZ) and height-for-age Z-score (HAZ) and weight-for-height Z-score (WHZ) between the intervention group and the control group (P>0.05). The rate of emaciation of the intervention group was 1.64%, which was significantly lower than 3.67% of the control group (P<0.05). There were no significant differences in the rate of growth retardation (2.81% and 3.32%, respectively) and underweight (0.47% and 1.92%, respectively) between the intervention group and the control group (P>0.05). The rate of respiratory infection and diarrhea in the intervention group were 9.13% and 1.17%, which were significantly lower than corresponding 23.25% and 3.15% in the control group (P<0.05). The hemoglobin of the intervention group and the control group were 117.24 g/L and 114.51 g/L respectively, and the rates of anemia were 11.11% and 22.84% respectively, the differences were statistically significant (P<0.05). Conclusions　The intervention of nutrition package in rural areas of Hainan Province has achieved the expected results, and supplementary food nutrition package has reduced the incidence of malnutrition and respiratory infection and diarrhea in recent two weeks in infants and anemia to a certain extent. We should attach great importance to the supplementary nutrition package for right-age children and promote the growth and health of children in rural areas through supplementary nutrition package, and continuously improve the nutrition and health level of children in Hainan Province.

OBJECTIVETo compare the efficacy and safety of docetaxol, pemetrexed and EGFR-TKIs in the second-line treatment for patients with advanced non-small cell lung cancer.

METHODSThe clinical data of 170 patients with advanced non-small cell lung cancer who failed standard first-line chemotherapy were reviewed. Those who received docetaxol as second-line therapy were designated as group A (n = 60), those who received pemetrexed as second-line therapy were designated as group B (n = 49), and those who received EGFR-TKIs as second-line therapy were designated as group C (n = 61). PFS and MST were estimated by Kaplan-Meier method and the differences between groups were compared by log-rank test.

RESULTSThe response rate in the groups A, B and C group was 15.0% (9/60), 24.5% (12/49) and 36.1% (22/61), respectively. The PFS after second-line therapy in the groups A, B and C was 5.49 months (95%CI: 4.03 - 6.95 months), 5.42 months (95%CI: 4.23 - 6.60 months) and 9.31 months (95%CI: 6.88 - 11.73 months), respectively (P = 0.045). The MST after second-line therapy in the groups A, B and C was 14.89 months (95%CI: 11.23 - 18.55 months), 15.81 months (95%CI: 12.11 - 19.52 months) and 17.47 months (95%CI: 13.38 - 21.56 months), respectively (P = 0.574). Regression analysis showed that the performance status score (PS) and response for second-line treatment are independent prognostic factors in each sub-group, and pathological type is an independent prognostic factor in the group C (P = 0.003).

CONCLUSIONSThe safety of the three drugs used as second-line treatment for patients with advanced non-small-cell lung cancer, who failed standard first-line chemotherapy, is comparable, but the EGFR-TKIs group has the highest response rate, and the EGFR-TKIs group has the longest PFS with a statistically significant difference, while there is no significant difference in MST among the three groups. When patients receive second-line treatment, the performance status < 2 and the response rate for second-line treatment are independent prognostic factors. Furthermore, pathological type (adenocarcinoma) is also an independent prognostic factor for EGFR-TKIs as second-line treatment.

Adenocarcinoma ; drug therapy ; pathology ; Aged ; Antimetabolites, Antineoplastic ; therapeutic use ; Antineoplastic Agents ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Disease-Free Survival ; Erlotinib Hydrochloride ; Female ; Glutamates ; therapeutic use ; Guanine ; analogs & derivatives ; therapeutic use ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; Male ; Neoplasm Staging ; Pemetrexed ; Protein Kinase Inhibitors ; therapeutic use ; Quinazolines ; therapeutic use ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; therapeutic use ; Survival Rate ; Taxoids ; therapeutic use