Objective:To explore the seizure recurrence and prognosis of epilepsy in relapse after anti-epileptic drugs (AEDs) withdrawal, and the influencing factors for these conditions.Methods:From December 2009 to August 2018, patients from the Affiliated Brain Hospital of Nanjing Medical University who relapsed after AEDs withdrawal were collected and followed up for at least 18 months. The seizure recurrence and prognosis of these patients were prospectively observed. The Kaplan-Meier method was used for survival analysis. The associated risk factors of the second relapse in the enrolled patients were analyzed by multivariate Cox analysis. The included patients were divided into good prognosis group and poor prognosis group according to whether they had achieved seizure freedom for at least one year after the first relapse. A multivariate Cox regression model was used to analyze the independent risk factors affecting their prognosis.Results:A total of 56 patients with epilepsy in relapse after AEDS withdrawal were collected. The average follow-up period was 46.23 months (18-120 months) from the initial time of seizure recurrence, and 21 patients (37.5%) had the second seizure recurrence. The relapsing risk in patients who continued to be observed without adding AEDs was higher than those who were treated immediately with drugs [9/16 vs 30.0% (12/40)], but without statistically significant difference (χ2=2.220, P=0.071). The results of univariate analysis showed that focal seizures, seizure frequency more than once per month before remission and poly-drug therapy before AEDs withdrawal were associated with high risk of the second relapse. Poly-drug therapy was an independent risk factor for the second relapse by multivariate Cox analysis ( HR=3.383, 95% CI 1.257-9.105). Of the 56 patients with epilepsy in relapse after AEDs withdrawal, 47 patients (83.9%) had a good prognosis without seizure for at least one year, and of 33 patients who were followed up for three years or more, 26 (78.8%) had no seizure for at least two years. Between the group retreated immediately after the first recurrence and the group without immediate treatment [87.5% (35/40) vs 12/16],there were no statistically significant differences on the proportions of good prognosis (χ2=2.333, P=0.258). Univariate analysis showed that the course of epilepsy>6 months before initial treatment, the frequency of seizures>1/month before remission, symptomatic epilepsy and poly-drug therapy were associated with the poor prognosis. However, none of independent risk factors was found for the poor prognosis through the multivariate analysis. Conclusions:The prognosis of patients with epilepsy in relapse after AEDs withdrawal is well, and about 2/3 patients with epilepsy in relapse after AEDs withdrawal have no more seizure recurrences. The poly-drug therapy before AEDs withdrawal may be an independent risk factor for the second seizure relapse.

The development of nano drug delivery systems (NDDSs) provides new approaches to fighting against diseases. The NDDSs are specially designed to serve as carriers for the delivery of active pharmaceutical ingredients (APIs) to their target sites, which would certainly extend the benefit of their unique physi-cochemical characteristics, such as prolonged circulation time, improved targeting and avoiding of drug-resistance. Despite the remarkable progress achieved over the last three decades, the understanding of the relationships between the in vivo pharmacokinetics of NDDSs and their safety profiles is insufficient. Analysis of NDDSs is far more complicated than the monitoring of small molecular drugs in terms of structure, composition and aggregation state, whereby almost all of the conventional techniques are inadequate for accurate profiling their pharmacokinetic behavior in vivo. Herein, the advanced bio-analysis for tracing the in vivo fate of NDDSs is summarized, including liquid chromatography tandem-mass spectrometry (LC-MS/MS), F(o)rster resonance energy transfer (FRET), aggregation-caused quench-ing (ACQ) fluorophore, aggregation-induced emission (AIE) fluorophores, enzyme-linked immunosor-bent assay (ELISA), magnetic resonance imaging (MRI), radiolabeling, fluorescence spectroscopy, laser ablation inductively coupled plasma MS (LA-ICP-MS), and size-exclusion chromatography (SEC). Based on these technologies, a comprehensive survey of monitoring the dynamic changes of NDDSs in struc-ture, composition and existing form in system (i.e. carrier polymers, released and encapsulated drug) with recent progress is provided. We hope that this review will be helpful in appropriate application methodology for investigating the pharmacokinetics and evaluating the efficacy and safety profiles of NDDSs.

Objective To explore if Ulinastai reverses hepatofibrosis in rats.Method Rat hepatofibrosis models were induced by TAA subcutaneously injection.18 rats with hepatofibrosis were divided into 2 groups,Ulinastai treatment group (9 rats),normal saline control group (9 rats).AST,ALT,HA,SOD,MDA,LN level were measured and compared between the 2 groups and that of healthy rats (3 rats).Rats liver morphology was observed using HE,Masson stain and type-B ultrasonic.TGF-β1、caspase-3 were detected by immunohistochemistry.Result After 3 weeks Ulinastai treatment,elastic index and SSS score in Ulinastai group decreased significantly compared to pretreatment (t =2.472,P ＜0.05).ALT,AST,SOD,MDA,HA and LN level significantly improved (F =3.862,5.774,3.442,4.157,4.173,3.674,P ＜ 0.05).Compared with NS treatment group HE and Masson staining showed fewer inflammatory cells infiltration in central vein,ballooning degeneration of liver cells,collagen proliferated,hepatic lobules degradation and pseudolobule arise after 3 weeks intervenion.Also in Ulinastai group,TGF-β1、caspase-3 positive cells were much less than that in NS treatment group.Conclusions Ulinastai can reverse rat hepatic fibrosis and alleviate fibrosis degree and collagen fiber deposit.

Objective The report was focused on polycystic ovary syndrome(PCOS) in female adolescents related to homosexual precocious puberty,and the new awareness of its influence on long-term health of homosexual precocious girls.Methods A girl with idiopathic central precocious puberty (ICPP) but diagnosed as PCOS in adolescence was reported and the data were reviewed.Results A girl was diagnosed as ICPP when she was near 8 years old with obesity,but without hyperinsulinemia,then she received the treatment of a 3.75 mg dose gonadotropin hormone analogues (GnRHa) every 28 days.Her gonads development was under control while her growth was arrested.Growth hormone(GH) injection started at 13 months.She stopped all medications when finished GH and GnRHa treatment for 12 months and 25 months.She went to see doctors again because of no menarche after discontinuing medication for nearly 2 years.She appeared obese and acne,hirsutism,athanens negricans and purple purple striae on the skin.Hyperinsulinemia and hypertestosterone were demonstrated.Pelvic B ultrasonography showed polycystic ovary,and she was diagnosed as PCOS.She was ordered to lose weight and to take metformin.And adrenocorticotropic hormone stimulating test was done,and B ultrasonography again ruled out atypical congenital adrenal hyperplasia and tumor of adrenal gland.She got her menarche 1 month later.Twelve months after the PCOS diagnosis and treatment,she had 4 menses,her insulin level decreased,glucose tolerance and her serum testosterone level turned normal.At the same time,the form of ovarian and follicular was significantly reduced.After taking mefformin for 20 months,her height and weight did not change,her menstrual was regular every 35-40 days,each time lasting about 7 days.She was followed up.We also reviewed literatures and learnt that precocious puberty might not be a benign situation and it might have an intrinsic relation to obesity,precocious puberty and PCOS.It might be the cofactors for causing PP and PCOS that accessed luteinizing hormone(LH) secretion and disorder of hypothalamus pituitary gonad(HPG) axis except obesity with hyperinsulinemia.Reported pubertas praecox in childhood developed to PCOS at 30％ with high prevalence of obesity.GnRHa suppressive therapy might relate to PCOS and had disputation for improving final height.There were fewer investigations on adolescent PCOS,and no consensus guideline on it in China.Conclusions Girls with ICPP may develop to PCOS some time later.The clinical features of the reported girl and the knowledge from literatures support the hypothesis that inner relationship between the CPP and the PCOS.The LH high secretion and disorder of HPG axis may be the causes of them; LH treatment on suppressing obesity should be cautious during treatment.The benefit and risk from GnRHa treatment should be evaluated thoroughly.Further clinical research should be conducted on adolescent PCOS.

The fragmentation pathways of five estrogens (estradiol, estrone, equilin sulfate, 17 a-dihydroequilin sulfate and equilenin sulfate) have been studied with high resolution and high mass accuracy using electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF/MS) in the negative ion mode. Molecular weights were obtained from [M-H](-) ions in the product ion spectra. The results indicate that the five structurally similar estrogens have similar fragmentation pathways. Using their stable isotope forms as internal reference compounds, the accurate mass and composition of the fragment ions were determined. During collision-induced dissociation (CID), cleavage is initiated by loss of oxygen atoms from carbon-17, after which D and C rings cleave sequentially and rearrange to finally form stable conjugate structures with highly abundant characteristic fragment ions at m/z 183 (accompanied by m/z 181), m/z 169 and m/z 145 (accompanied by m/z 143). Understanding these characteristic fragmentation pathways of estrogens will be helpful in identifying the structures of steroid hormones in general.

OBJECTIVE:To study the pharmacokinetics of balofloxacin tablets in Chinese healthy volunteers.METHODS:A single dose of 100 or 200 mg balofloxacin tablets were given to 12 healthy volunteers in a randomized crossover design.Concentrations of balofloxacin in plasma and urine were determined by HPLC with data processed by DAS(drug and statistics)software.RESULTS:After administration of balofloxacin 100 mg and 200 mg,the Cmax were(0.970?0.245)?g?mL-1 and(1.849?0.466)?g?mL-1;the tmax were(1.25?1.10)h and(1.24?0.81)h;the t1/2 were(7.14?1.01)h and(7.11?0.72)h;the AUC0～36 were(7.309?1.368)?g?h?mL-1 and(15.214?1.727)?g?h?mL-1;AUC0～∞ were(7.531?1.386)?g?h?mL-1 and(15.695?1.762)?g?h?mL-1;the accumulative eliminating rates within 36 h in urine were(64.47?11.56)% and(63.24?11.93)%,respectively.CONCLUSION:The pharmacokinetics of balofloxacin in healthy volunteer after oral administration was characterized by high peak concentration and long half life.The method is sensitive,accurate,reliable and specific,and can satisfy the requirements for pharmaceutical study.

AIM: To determine the relationship between the mutation of the RII gene and RER status in the tumorigenesis of sporadic colorectal cancer. METHODS: We screened RER status and mutation of the RII gene from 50 sporadic colorectal cancers (19 in the proximal colon, 31 in the distal colorectum). RESULTS: RER was found in 13 cases (8 in the proximal colon, 5 in the distal colorectum), and 5 of them showed mutations of the RII gene. All 5 cancers carrying a TGF-? RII gene mutation showed RER+, but there wasn't any mutation of RII gene in RER(-) cases. Four of 5 RII mutation were located at the cecum. CONCLUSION: These data indicate that the TGF-? RII gene is a major target of microsatellite instability and mutation of the RII gene play an important role in carcinogenesis of sporadic colorectal cancer with microsatellite instability, especially at the cecum. [

Objective: To better understand the function of mast cells in acupuncture points (acupoints) inacupuncture-induced analgesia. The author tested their sensitivity to mechanical, thermo and light stimulation.Methods: The tail flick model was applied to measure analgesia in rats, and the author determined the density of mast cells in tissue slices and their degranulation ratio before/after acupuncture. The author also applied the patch-clamp technique to investigate activation of human mast cells (HMC 1 cell line) by mechanical stress or noxious heat, and the author optically observed degranulation phenomena of mast cell in response to red laser light.Results: Manual stimulation by acupuncture at Zusanli (ST 36) of the rat resulted in analgesia and the effect was more pronounced than after stimulation of a sham point nearby the acupuncture point. A higher density of mast cells was found at Zusanli (ST 36) than at the sham point, and acupuncture caused a remarkable increase in degranulation. Pretreatment with disodinm chromoglycate (DSCG, a mast cell stabilizer) injected into Zusanli (ST 36) impaired the analgesic effect of acupuncture. In whole-cell patch-clamp, increasing hydrostatic pressure induced a current that could be inhibited by 10 ～tM of ruthenium red (RuR), an inhibitor of TRPV2. Temperatures above 50～C, which are reached during moxibustion, induced a similar RuR-sensitive current. Laser light of 640 nm (48 mW) applied to acupoints had been shown previously to be highly effective in analgesia. 20 min of light application induced pronounced degranulation in HMC 1 that could be blocked by 0.02 g/mL DSCG as well as by RuR. Conclusion: The author found that mechanical stimulation of acupoints is associated with mast cell degranulation and mast-cell degranulation can be correlated with acupuncture-induced analgesia. The author suggest that TRPV2 plays a key role in mast-ceU degranulation in response to mechanical, thermal (moxibustion) as well as laser-light stimulation forming a facilitating step in acupuncture-induced analgesia.

OBJECTIVETo study the effect of ginsenoside Rb1 on GSKbeta/IDE signal transduction pathway and Abeta protein secretion in hippocampal neurons of high glucose-treated rats.

METHODHippocampal neurons of 24 h-old newly born SD rats were primarily cultured, inoculated in culture medium under different conditions, and then divided into the normal group, the high glucose group, the LiCl group and the Rb1 group. After being cultured for 72 h, the expressions of their phosphorylated GSK3beta, total GSK3beta and IDE protein were detected by Western blotting analysis. The mRNA expressions of GSK3beta and IDE were determined by RT-PCR. The ELISA assay was used to detect the secretion of Abeta protein in cell supernatant.

RESULTCompared with the normal group, the high glucose group showed increase in the p/tGSK3beta protein ratio and the secretion of Abeta protein and decrease in IDE protein and mRNA (P < 0.05). Compared with the high glucose group, both Rb1 and LiCl groups showed decrease in the p/tGSK3beta protein ratio and the expression of Abeta protein and increase in IDE protein and mRNA expression (P < 0.05). Compared with the LiCl group, the Rb1 group showed no significant difference in the expressions of p/tGSK3beta protein, IDE protein, mRNA and Abeta protein expression. In addition, the GSK3beta mRNA expression of the four groups had no significant difference.

CONCLUSIONGinsenoside Rb1 may reduce the secretion of Abeta protein in hippocampal neurons by reducing the phosphorylation of GSK3beta, down-regulating the ratio of pGSK3beta/GSK3beta and upregulating the expression of IDE.

Amyloid beta-Peptides ; genetics ; metabolism ; secretion ; Animals ; Dietary Carbohydrates ; adverse effects ; Gene Expression Regulation ; drug effects ; Ginsenosides ; pharmacology ; Glucose ; adverse effects ; Glycogen Synthase Kinase 3 ; genetics ; metabolism ; Glycogen Synthase Kinase 3 beta ; Hippocampus ; cytology ; Insulin ; metabolism ; Insulysin ; genetics ; metabolism ; Neurons ; cytology ; drug effects ; metabolism ; secretion ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects

The Doha Declaration has entrusted to the members of WHO the right to use the compulsory licensing for drug patents in the face of global public health crises.Although the basic compulsory licensing system for drug patents has been established in China, the efficiency of its implementation is low, the responsibility of its executive departments is unclear, the related rules are not fully understood by pharmaceutical enterprises, it is thus necessary to improve and perfect the relevant legal provisions and systems in order to effectively improve the accessibility of drugs.