Borneol is a traditional Chinese medicine. In the past few years, many studies showed that borneol can improve the bioavailability of other drugs, promoting drugs to cross the blood-brain barrier, so the potential drug interactions between borneol and other medicines have attracted great attention, but the influence of borneol to CYP450 and its isoforms are rarely reported. In this research, male Wistar rats were orally administered by borneol for 7 days, then the mRNA and protein expression and the activities of CYP2D were detected, we also compared the pharmacokinetic parameters of CYP2D's specific substrate between control group and borneol group. The results show that borneol (33, 100 and 300 mg x kg(-1) x d(-1)) have no significant effect on CYP2D, while the activites of CYP2D increased 1.71, 1.97 and 2.89 times comparing to the control group. At the same time, borneol (300 mg x kg(-1) x d(-1)) caused the C(max) decreased 10.6% (P > 0.05), AUC(0-∞) decreased 27.5% (P < 0.01), CL/F increased 41.1% (P < 0.01), V(z)/F increased 23.1% (P > 0.05) of dextromethorphan. Our data provided that borneol speed up dextromethorphan's elimination in vivo. Since the activity of CYP2D can be induced by borneol, the metabolic interactions might happen when borneol and the substrate drug CYP2D are used together.

Objective:To explore the mechanism of promotion effect of juglone combined with cisplatin on the apoptosis of human cervical cancer HeLa cells,and to clarify the effects of its associated signal transduction pathways.Methods:The HeLa cells at logarithmic growth phase were divided into control group,juglone group, cisplatin group and juglone combined with cisplatin group (combined treatment group).The inhibitory rates of proliferation of HeLa cells were detected by MTT assay.The apoptosis was detected by Hoechst 33258 staining. The expressions of Bcl-2, Bax and caspase-3, AKt, and pAKt were detected by Western blotting method. Results:The MTT results showed that the HeLa cell proliferation at 24,48 72 h in each drug group was inhibited;compared with control group,the profileration of HeLa cells in juglone group and cisplatin group was significantly inhibited,especially in combined group. Compared with single drug group,the inhibitory effect in combined treatment group was more significantly.After treatment for 12 h,the typical morphological changes of apoptosis were found in juglone group and cisplatin group by Hoechst 33258 staining,especially in combined treatment group. The Western blotting results showed that the expression levels of Bcl-2 and pAKt in HeLa cells in juglone group and cisplatin group 12 h after treatment were decreased obviously,whereas the expression levels of Bax,Caspase-3,and AKt were increased significantly, especially in combined treatment group compared with control group. Conclusion:Juglone combined with cisplatin could inhibit the PI3K/AKt pathway,thereby promoting the apoptpsis of HeLa cells.

AIM:To explore the effect of peptidyl-prolyl cis/trans isomerase (Pin1) inhibitor juglone on apop-tosis of human cervical cancer SiHa cells.METHODS:Cultured SiHa cells were incubated with juglone at concentrations of 10, 20, 50, 80 and 100 μmol/L for 24 h.The SiHa cell activity was detected by methyl thiazolyl tetrazolium ( MTT) assay.The cell apoptosis was analyzed by flow cytometry with Hoechst 33258 staining.The protein levels of cleaved caspase-3,8,9 and PTEN was determined by Western blotting.RESULTS:In different doses of juglone groups, the SiHa cell growth was greatly inhibited ( P<0.05) in a dose-dependent manner as compared with control group.The IC50 of ju-glone was 20.4 μmol/L.After treatment with juglone at concentration of 20 μmol/L for 12 h, the apoptosis of SiHa cells was induced, and the typical morphological changes of cell apoptosis such as karyopyknotic pyknic hyperfluorescence bolus, nuclear fragmentation and apoptotic body were observed by Hoechst 33258 staining.The early apoptotic rate was increased significantly as compared with the control.The protein levels of cleaved caspase-3, 8, 9 and PTEN were also increased sig-nificantly as compared with control group.CONCLUSION:Juglone significantly inhibits the cell activity and induces the apoptosis of SiHa cells in vitro by inhibiting the caspase pathway and increasing the expression of anti-oncogene.

BACKGROUND: Considerable studies demonstrate that nitric oxide synthase (NOS)/nitric oxide (NO)plays an important role in maintaining normal function of Sertoli cells, and influences spermatic generation and activation as well as fertilizability.OBJECTIVE: To observe the effect of goat testis extract on NOS activity in Sertoli cells of mice with testis injury caused by heavy mental Pb.DESIGN: Randomized controlled animal trial.SETTING: Department of Histology and Embryology, Jilin Medical College.MATERIALS: This trial was carried out in the laboratory of Histology and Embryology, Jilin Medical College (Key laboratory of the general logistics department of P.L.A) during March 2004 to August 2005. Thirty healthy Kunming male mice were involved and randomized into 3 groups,with 10 in each group: control group, testis injury model group (model group) and goat testis extract-treated group (treatment group).METHODS: The mice in the model group and experimental group were daily administrated with 100 g/L lead acetate, 0.2 mL/(mouse·d), 5 times/wk within 2 weeks, then withdrawal for 1 week. Simultaneously, the mice in the treatment group were subcutaneously injected with goat testis extract at 0.5 mL/(mouse ·d). The mice in the control group were given redistilled water of the same dose as that in the treatment group. After being poisoned fully, the mice were fasted for 12 hours and weighted, finally sacrificed by decapitation. Bilateral testis were dissected, immediately weighted, fixed with formalin and sliced. The NOS changes in Sertoli cells of mice in each group were observed with reduced nicotinamide-adenine dinucleotide phophate-diaphorase(NADPH-d) histochemical method combined with microscope image.MAIN OUTCOME MEASURES: Body mass, bilateral testis mass and NOS absorbance (A) in Sertoli cells of mice in each group after contamination expiration.RESULTS: All the 30 mice were involved in the result analysis, without deletion. ①After contamination expiration, the body mass, bilateral testis mass and NOS A value in treatment group were significantly than those in the model group [(22.47±3.49) g vs. (19.13±3.46) g;(0.113±0.021 ) g vs.(0.089±0.017) g; 0.236±0.020 vs. 0.146±0.023, t=2.151-3.314,P ＜ 0.05-0.01]; ② In the model group, NOS positive Sertoli cells swelled and degenerated; The morphology of NOS positive Sertoli cells in the treatment group was close to that in the control group.CONCLUSION: Goat testis extract can boost the NOS activity in Sertoli cells of mice with testis injury caused by heavy mental pliumbum and has some repairing and protective effect on testis injury, which can provide new thinking for treatment of male sterility.

Objective To identify the anatomic advantages of pre-contouring long helical plate of proximal humerus internal locking system (PHILOS) aided by 3D-printing technique in minimally invasive plate oseoynthesis (MIPO) for metadiaphyseal fractures of the proximal humerus.Methods Firstly in a cadaveric study,12 fresh frozen samples of upper extremity were divided into 2 equal groups:6 in a Synbone group and 6 in a 3D-printing group.Long 10-hole PHILOS plates were pre-contoured helically according to Synbone humerus models in the Synbone group but according to 3D-printing humerus models in the 3D-printing group.All these helical plates were fixed onto the cadaveric humerus using MIPO technique respectively.The horizontal distances between the radial nerve and the lateral side of the plate were measured at the point where the radial nerve penetrates the muscular septum,at the 6th hole on the plate and at the distal end of the plate;the distance between the medial side of the distal plate and the upper arm neurovascular bundle was also measured.Finally,the full lengths of all these cadaveric humeri were measured.Secondly,on the basis of the anatomic study,16 patients with metadiaphyseal fracture of the proximal humerus were treated at Department of Orthopedic Surgery,The Sixth People's Hospital of Shanghai from February 2013 to March 2016,using the same MIPO technique and the same long 10-hole PHILOS plates pre-contoured helically on the 3D printing models which were mirrored on the humerus on the healthy side of the patients.They were 4 men and 12 women,aged from 59 to 87 years (mean,71.0 years).By the AO/OTA classification,10 cases were type B and 6 type C.The fracture line involved the proximal humerus in 7 cases.Postoperative complications were followed up.At the final follow-up,the shoulder function was assessed according to the Constant-Murley score and the elbow function according to the Mayo Elbow Performance Score (MEPS).Results By the anatomic measurement,the full length of cadaveric humerus averaged 30.17 ± 1.93 cm in the Synbone group and 29.75 ± 2.17cm in the 3D-printing group,showing no significant difference (P ＞ 0.05).The horizontal distances between the radial nerve and lateral side of the plate at the point where the radial nerve penetrates the muscular septum,at the 6th hole on the plate and at the distal end of the plate in the Synbone and 3D-printing groups were,respectively,11.50 ± 0.92 mm versus 17.87 ± 1.88 mm,6.90 ± 1.78 mm versus 14.83 ± 1.50 mm,and 5.14 ± 1.14 mm versus 8.36 ± 1.27 mm,all showing significant differences between the 2 groups (P ＜ 0.05％).The distance between the medial side of the distal plate and the upper arm neurovascular bundle was 6.25 ± 0.95 mm in the Synbone group and 6.88 ± 1.21 mm in the 3D-printing group,showing an insignificant difference between the 2 groups (P ＞ 0.05).The clinical observation found no iatrogenic nerve injury.The 16 patients were followed up for an average of 25.3 months (range,from 24 to 38 months).All fractures got united uneventfully.At the final follow-up,their Constant-Murley scores averaged 77.6 points and their MEPS 96.5 points.Conclusion The risk of iatrogenic injury to the radial nerve can be lowered when 3D-print technique is applied for helical pre-contouring of long PHILOS plate in the MIPO for metadiaphyseal proximal humeral fractures,leading to satisfactory clinical results.

BACKGROUNDIn vivo quantification of choroidal neovascularization (CNV) based on noninvasive optical coherence tomography (OCT) examination and in vitro choroidal flatmount immunohistochemistry stained of CNV currently were used to evaluate the process and severity of age-related macular degeneration (AMD) both in human and animal studies. This study aimed to investigate the correlation between these two methods in murine CNV models induced by subretinal injection.

METHODSCNV was developed in 20 C57BL6/j mice by subretinal injection of adeno-associated viral delivery of a short hairpin RNA targeting sFLT-1 (AAV.shRNA.sFLT-1), as reported previously. After 4 weeks, CNV was imaged by OCT and fluorescence angiography. The scaling factors for each dimension, x, y, and z (μm/pixel) were recorded, and the corneal curvature standard was adjusted from human (7.7) to mice (1.4). The volume of each OCT image stack was calculated and then normalized by multiplying the number of voxels by the scaling factors for each dimension in Seg3D software (University of Utah Scientific Computing and Imaging Institute, available at http://www.sci.utah.edu/cibc-software/seg3d.html). Eighteen mice were prepared for choroidal flatmounts and stained by CD31. The CNV volumes were calculated using scanning laser confocal microscopy after immunohistochemistry staining. Two mice were stained by Hematoxylin and Eosin for observing the CNV morphology.

RESULTSThe CNV volume calculated using OCT was, on average, 2.6 times larger than the volume calculated using the laser confocal microscopy. The correlation statistical analysis showed OCT measuring of CNV correlated significantly with the in vitro method (R 2 =0.448, P = 0.001, n = 18). The correlation coefficient for CNV quantification using OCT and confocal microscopy was 0.693 (n = 18, P = 0.001).

CONCLUSIONSThere is a fair linear correlation on CNV volumes between in vivo and in vitro methods in CNV models induced by subretinal injection. The result might provide a useful evaluation of CNV both for the studies using CNV models induced by subretinal injection and human AMD studies.

Animals ; Choroidal Neovascularization ; pathology ; physiopathology ; Disease Models, Animal ; Fluorescein Angiography ; Humans ; Mice ; Mice, Inbred C57BL ; Tomography, Optical Coherence

INTRODUCTIONRetinitis pigmentosa (RP) is the most prevalent group of inherited retinopathies and demonstrates considerable clinical and genetic heterogeneity, with wide variations in disease severity, progression, and gene involvement. We studied a large family with RP to determine the pattern of inheritance and to identify the disease-causing gene/locus.

MATERIALS AND METHODSOphthalmic examination was performed on 35 family members to identify affected individuals and carriers and to characterise the disease phenotype. Genetic linkage analysis was performed using short tandem repeat (STR) polymorphic markers encompassing the known loci for Xlinked RP (xlRP) including RP2, RP3, RP6, RP23, and RP24. Mutation screening was performed by direct sequencing of PCR-amplified genomic DNA of the RP2 and RPGR genes of the affected individuals.

RESULTSA highly penetrant, X-linked form of RP was observed in this family. Age of onset was from 5 to 8 years and visual acuity ranged from 20/25 in children to light perception in older adults. Linkage analysis and direct sequencing showed that no known loci/genes were associated with the phenotype in this kindred.

CONCLUSIONA novel disease gene locus/loci is responsible for the xlRP phenotype in this family.

Adolescent ; Adult ; Age of Onset ; Child ; Child, Preschool ; Chromosome Mapping ; DNA Mutational Analysis ; Eye Proteins ; genetics ; Female ; Genetic Diseases, X-Linked ; genetics ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Lod Score ; Male ; Membrane Proteins ; genetics ; Pedigree ; Phenotype ; Retinitis Pigmentosa ; genetics

Background Global warming may increase the frequency of compound hot extreme (CHE).However, there is still a lack of studies assessing the associations between CHE and preterm birth (PTB), and the underlying biological mechanisms remain unclear. Objective To estimate the association of exposure to CHE during pregnancy with PTB, and to explore the roles of inflammatory, endothelial dysfunction, and oxidative stress in the association between CHE and PTB. Methods All participants were selected from the Prenatal Environments and Offspring Health (PEOH), a prospective birth cohort conducted in Guangzhou. In this study, a total of 2449 participants who gave birth from May to October in 2014 to 2017 were enrolled, and among them blood samples were collected from 311 preterm (n=43) and full-term (n=268) pregnant women at the time of delivery. A hot day/night was identified as a day when the daily maximum temperature/minimum temperature was higher than its 90th percentile in the study period, and a CHE was defined as having both a hot night and a following hot day. The meteorological data were obtained from the China Meteorological Data Sharing Service System. Anusplin was used to assess the daily maximum temperature, daily minimum temperature, and relative humidity of the participant residence. Enzyme-linked immunosorbent assay (ELISA) was used to measure C reactive protein (CRP), endothelin-1 (ET-1), and malondialdehyde (MDA) levels in maternal serum, and their results were transformed by natural logarithm. A distributed lag nonlinear model was used to investigate the associations of exposures to hot day, hot night, and CHE during pregnancy with PTB at different lag days, and a logistic regression model was used to investigate the associations of CRP, ET-1, and MDA with PTB. Results The incidence rate of PTB was 6.2% in all selected participants. Compared with the non-hot day, the RRs (95%CIs) of CHE in lag 3, 7, and 14 days on PTB were 1.43 (1.12-1.84), 1.24 (1.08-1.43), and 1.17 (1.05-1.30), respectively, and the cumulative effects (% difference) (95%CI) of CHE in lag 14 days on maternal serum CRP, ET-1, and MDA were 0.33% (−0.45%-1.12%), 0.59% (0.11%-1.07%), and 0.57% (0.09%-1.05%), respectively. Compared with the Q1 (lowest quartile) for CRP, ET-1 and MDA, the RRs (95%CIs) of Q4 (highest quartile) for PTB were 1.27 (0.50-3.22), 1.51 (0.61-3.72), and 2.07(0.81-5.27), respectively. Conclusion Maternal exposure to CHE during pregnancy might be associated with an increased risk of PTB. Prenatal exposure to CHE is positively associated with maternal serum CRP, ET-1, and MDA, and the three biochemical indicators are also positively associated with PTB. However, the above conclusions still need further confirmation.