OBJECTIVETo study the effect of Guishen Pill (GSP) on expression levels of Oct-4, MVH, and Egr-1 in mice with diminished ovarian reserve (DOR).

METHODSTotally 40 female C57BL/6J mice were randomly divided into 4 groups, the normal control group, the model group, the GSP group, and the dehydroepiandrosterone (DHEA) group, 10 in each group. Pregnant mare serum gonadotropin (PMSG), human chorionic gonadotropin (HCG), and prostaglandin F2α (PGF2α) were sequentially administrated to produce superovulation. The DOR model was established by exposing to ozone inhalation. Mice in the GSP group were intragastrically administered with GSP at 0.3 mL. Those in the DHEA group were intragastrically administered with DHEA at 0.3 mL. Equal volume of normal saline was intragastrically administered to mice in the normal control group and the model group. All mice wer treated for 21 days. Serum levels of estrogen (E2), progestogen (P), and anti-Müllerian hormone (AMH) were measured by ELISA. Changes of Oct-4, anti-AMH, and early growth response gene-1 (Egr-1) mRNA in ovaries were dtected by Real-time PCR.

RESULTSCompared with the model group, serum levels of E2, P, and AMH, as well as contents of estrogen receptor (ER), progestogen receptor (PR), MVH, and Oct-4 mRNA significantly increased in the GSP group and the DHEA group (P < 0.05).

CONCLUSIONGSP could improve expression levels of Oct-4, MVH, and Egr-1 mRNA in DOR mice and their ovarian function.

Animals ; Anti-Mullerian Hormone ; metabolism ; Dehydroepiandrosterone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Early Growth Response Protein 1 ; metabolism ; Estrogens ; Female ; Mice ; Mice, Inbred C57BL ; Octamer Transcription Factor-3 ; metabolism ; Ovarian Reserve ; Ovary ; Pregnancy ; Receptors, Estrogen ; metabolism ; Superovulation

Background Epithelial-mesenchymal transition (EMT) is a major event in the pathogenesis of posterior capsular opacification (PCO),and the expressions of α-smooth muscle actin (α-SMA) is the marker of EMT.Previous studies showed that breviscapine plays an important role in anti-fibrosis and suppression EMT,however,the mechanism of its effect on EMT in LECs is unclear.Objective Present study was to investigate the effect of breviscapine on the expression of α-SMA and fibronectin (FN) in human LECs induced by transforming growth factor-β2 (TGF-β2).Methods Human LECs strain,HLE-B3,was cultured and passaged in DMEM containing 10％ fetal bovine serum.Different concentrations of breviscapine (6.75,12.75,25.00,50.00 and 100.00 mg/L)were added into the medium for 24,48 and 72 hours respectively,and then cell cunting kit-8 (CCK-8) was used to evaluate the half maximal inhibitory concentration (IC50) of breviscapine to HLE-B3.In addition,HLE-B3 was subcultured at the density of 1 × 106/hole and divided into 4 groups.The cells were in free-serum medium as the normal control;the cells were exposed in 10 μg/L TGF-β2 as TGF-β2 group;while in the breviscapine group,10 mg/L of breviscapine was added into the culture medium and another group was the combination of 10 mg/L breviscapine and 10 μg/L of TGF-β2 treatment.Real-time PCR and Western blot were used to detect the mRNA expressions of α-SMA and FN as well as their protein in HLE-B3 72 hours after cultured.Results The inhibitory rate of breviscapine to HLE-B3 proliferation was gradually elevated with the increase of concentration of breviscapine,showing a significant inhibition of the cell proliferation among the different groups and at various time points (F =292.851,P=0.000;F =65.037,P=0.000).IC50 of HLE-B3 at 72 hours was 22 mg/L,and therefore,in rest of experiment 10 mg/L of breviscapine was used which was 1/2 only half of the IC50.α-SMA and FN were expressed in cultured normal HLE-B3.The expressing level of α-SMA mRNA and FN mRNA in the HLE-B3 was significantly different among the normal group,TGF-β22 group,breviscapine treatment group and combination group as well as at various time points (F =105.490,P =0.000 ; F =1041.414,P =0.000).Similarly,the protein expressions of α-SM A and FN in the HLE-B3 was significantly different among the four groups and different time points (F=136.872,P=0.000;F=119.820,P=0.000).The expression levels of α-SMA and FN mRNA and their proteins in HLE-B3 were remarkably increased in the 10 μg/L TGF-β2 group compared with the normal control group (at all P=0.000),and those in the combination group were obviously declined in comparison to the TGF-β2 group (P =0.001,0.001,0.001,0.010).No significant difference was found in the expressions of α-SMA and FN in the HLE-B3 between the breviscapine group and normal control group in both transcriptional level and protein level (P =0.551,0.292,0.551,0.360).Conclusions 10 mg/L breviscapine can arrest the proliferation and EMT of human LECs.This result suggests that using breviscapine may be a potential prophylactic approach in the prevention of PCO.

BACKGROUND:Large numbers of experimental data have confirmed that bone marrow mesenchymal stem cel s have a positive therapeutic effect on cerebral ischemia-reperfusion injury, but there are few reports about intravenous administration of bone marrow mesenchymal stem cel conditioned medium in the treatment of stroke.
OBJECTIVE:To investigate the effects of the conditioned medium of rat bone marrow mesenchymal stem cel s on the recovery of neurological function in rats after cerebral ischemia-reperfusion injury.
METHODS:The bone marrow mesenchymal stem cel s were isolated from rat bone marrow. When cel s at passage 2 or 3 reached 90%confluence, the original culture medium was removed. Then the cel s were cultured in serum-free DMEM for 18 hours. After that, the culture solution was col ected as the conditioned medium of rat bone marrow mesenchymal stem cel s. Adult rats were subjected to 2 hours of right middle cerebral artery occlusion. Ischemia-reperfusion injury rats were randomly assigned to three groups:control group, simple culture medium group and conditioned medium group, and respectively given injection of normal saline, DMEM, conditioned medium (10 mL/kg) via the tail vein at 2, 24, 48 hours after operation.
RESULTS AND CONCLUSION:There was no difference in the behavioral tests among the three groups at postoperative 2 hours (P>0.05). Compared with the control and simple culture medium group, neurological impairment was significantly improved in the conditioned medium group at postoperative 1, 3, 5 days (P<0.05), but there was no significant difference between the control and simple culture medium groups. At postoperative 5 days, brain edema was significantly eased in the conditioned medium group in comparison to the control and simple culture medium groups (P<0.05), and there was also no difference between the latter two groups (P>0.05). These results suggest that rat bone marrow mesenchymal stem cel s-conditioned medium via intravenous administration can significantly ease brain edema and improve the neurologic function after cerebral ischemia-reperfusion injury.

2,4-Dichlorophenoxyacetic Acid ; analogs & derivatives ; toxicity ; Animals ; Epididymis ; growth & development ; metabolism ; Female ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Herbicides ; toxicity ; Male ; Rats ; Rats, Wistar ; Spermatozoa ; drug effects ; growth & development ; metabolism

AC133 Antigen ; Adult ; Aged ; Antigens, CD ; metabolism ; Carcinoma, Hepatocellular ; diagnosis ; metabolism ; pathology ; Female ; Glycoproteins ; metabolism ; Humans ; Liver Neoplasms ; diagnosis ; metabolism ; pathology ; Male ; Middle Aged ; Peptides ; metabolism ; Prognosis ; Thy-1 Antigens ; metabolism

Based on variation of Pinus massoniana families, heritablility and correlation analysis, the contents of shikimic acid and procyanidine (heritability 0.90, 0.70), dry weight of single branch (heritability 0.60) and and leaf length (heritability 0.46) were screened out as quality, yield and harvest cost traits of Folium Pini, respectively. For the different medicinal application of Folium Pini, varied methods were chosen to estimate weight and construct index equation. Weight adjustment based.on equal emphasis were used as economic weight determining method to select the best families, and the index (accuracy 0. 936 4 and heritability 0. 881 6) obtained was a little better than that obtained by equal emphasis, and much better than that by restricted index. The superior families selected with adjustment weight and equal emphasis were No. 46, 43 and 28. Partial regression were used as economic weight determining method to select the best families,and the index obtained had the highest accuracy (0.941 5) , index heritability (0. 889 9) and the genetic gain of shikimic acid content. The superior families selected with this method were No. 46, 27 and 47. No. 46 was the best families with maximal economic benefit. Our study indicated that suitable method for estimate weight and construct index equation can be applied for better accuracy of superior families selection of P. massoniana.
Breeding ; Drugs, Chinese Herbal ; analysis ; Pinus ; chemistry ; classification ; genetics ; growth & development ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; classification ; genetics ; growth & development

BACKGROUNDMost HCC patients with decompensation of liver function lost the chance of surgical and/or interventional treatment. The aim of this study was to evaluate feasibility and outcome of radiofrequency ablation (RFA) in treating hepatocellular carcinoma (HCC) patients with poor liver function (Child-Pugh class C), who are not suitable for surgery or hepatic artery chemo-embolization.

METHODSThirteen HCC patients (the number of tumors was 17) with liver function of Child-Pugh C (scores: 10.2 +/- 0.4) were included in the study. Among the patients, 8 were male and 5 were female with the average age of (61.6 +/- 10.9) years old. The average size of HCC was (3.8 +/- 1.0) cm. Two patients were recurrent HCC and 30.8% of the patients had multiple tumors (2 - 3 tumors). All the patients were treated with RFA.

RESULTSThere were 22 RFA sessions (1 - 4 sessions per patient) in all, average ablations per tumor at first session was 3.1. One week after RFA, the liver enzymes elevated in 9 patients (69.2%), in 7 of them, the liver enzyme returned to pre-RFA level in 1 - 3 months. One month after RFA, the Child-Pugh grading was 10.3 +/- 0.8 (Child-Pugh C), while that of pre-RFA was 10.2 +/- 0.4 (Child-Pugh C), with no significant difference. Computer tomography (CT) one month after RFA showed that the tumor necrosis rate was 88.2% (15/17). Five patients had 2 - 4 repeated RFA due to HCC recurrence. During the follow-up of 2- 69 months in this group, survival rate of one year was 53.8%, two years was 30.8%, and three year was 15.4%. The incidence of RFA-related complications was 13.6% (3/22 sessions), including 1 case of GI hemorrhage and 1 sub-capsular hemorrhage of the liver. One patient with HCC over 5 cm who had fever and liver abscess after RFA, and was dead 2 months later due to liver function failure.

CONCLUSIONSMinimal invasive RFA provides possible treatment modality for HCC patients with poor liver function, who are not candidates for surgical and/or interventional therapy. For large HCC, due to the required extended treatment region, special attention should be paid to the possibility of acute liver failure.

Adult ; Aged ; Carcinoma, Hepatocellular ; therapy ; Catheter Ablation ; methods ; Female ; Humans ; Liver Cirrhosis ; therapy ; Liver Neoplasms ; therapy ; Male ; Middle Aged ; Treatment Outcome

Objective To develop a new algorithm to reconstruct the distribution of acoustic sources of magnetoacoustic tomography with magnetic induction(MAT-MI)in the acoustic inhomogeneous media,which is developed on the basis of generalized finite element method (GFEM) and modified time inversion algorithm. Methods The acoustic and acoustic coupling theory and the basic equations of acoustics were used to study the forward and inverse problems of the acoustic inhomogeneous concentric sphere magneticacoustic coupling model. The solution of acoustic non-uniform media wave equation based on GFEM was proposed.The method solved the problem of acoustically inhomogeneous media sound source reconstruction and conductivity reconstruction.At the same time,the distribution of velocity was reconstructed by rotating the pairs of transducers and the time reversal algorithm. Results The proposed algorithm could accurately reconstruct the acoustic source distribution in acoustic inhomogeneous media,and could obtain the distribution of sound velocity during the reconstruction of sound source and recover the image well. Conclusion The proposed algorithm had its feasibility and effectiveness verified,and gains advantages in MAT-MI reconstruction of acoustic inhomogeneous media.

This study was aimed to explore the influence of recipient age on the occurrence of acute graft-versus-host disease (aGVHD) in mice. 8 - 10 weeks aged C57BL/6 (H-2K(b)) mice were selected as donors, 18 - 20 weeks aged and 8 - 10 weeks aged BALB/c (H-2K(d)) mice were served as recipients. 18 - 20 weeks and 8 - 10 weeks aged mice were all randomly divided into three groups: normal control group (without any treatment); irradiation alone group [administered a total body irradiation (TBI) without bone marrow transplantation] and model group [infused with bone marrow mononuclear cells 5 × 10(6) and splenocytes 5 × 10(5) from donor C57BL/6 (H-2K(b)) mice through caudal vein no more than 4 h after TBI]. The general state and survival rate of all mice were observed everyday. The factors (the chimerism in peripheral blood, T lymphocyte and their subsets, the percentage of Th1 cells) of mice in model groups were measured by flow cytometry on day 5, 10, 15, 20, 25, 30 after TBI, the leukocytes in peripheral blood were also calculated by direct microscopic counting. The histological examinations of liver, intestine and skin were done by hematoxylin and eosin staining on day 5, 15, and 25 after TBI. All above data were compared between model groups. The results indicated that murine model with aGVHD was established in two model groups. Compared with 8 - 10 weeks aged mice, the 18 - 20 weeks aged mice showed higher survival rate and lower clinical scores (P < 0.05); the reconstitution time of leukocyte and chimerism in peripheral blood were delayed (P < 0.05); The ratio of CD8(+)T lymphocytes and Th1 cells in peripheral blood were lower (P < 0.05); the histological changes of liver, intestine and skin were little. It is concluded that 18 - 20 weeks aged recipient mice exhibited a lower incidence of aGVHD than 8 - 10 weeks aged recipient mice.
Age Factors ; Animals ; Bone Marrow Transplantation ; methods ; Graft vs Host Disease ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transplantation, Homologous

OBJECTIVETo discuss and analyze the feasibility and strategy for perform the newborn gene screening in the process of newborn hearing screening in order to supply the defects or limitation in the hearing screening.

METHODSFour hundreds and sixty newborn babies from December 2006 to April 2007 accepted the simultaneous hearing and gene screening. Otoacoustic emission (OAE) was used for the first step hearing screening and OAE combined with auto auditory brainstem response (AABR) detection for the second step screening. Newborn genetic disease screening cards were used for collecting the blood spot from the umbilical cord within the moment of newborn. The cards could be directly performed the polymerase chain reaction (PCR) for screening the mitochondrial 12SrRNA 1555G and GJB2 as well as SLC26A4 genes mutations. The restriction enzyme Alw26I was used to recognize the point mutation of 12SrRNA A1555G. The samples with the possible 12SrRNA A1555G mutation were then sequenced to verify. The PCR products from the GJB2 coding region and SLC26A4 IVS7-2A > G hot spot region were sequenced directly. The software of DNAStar was used to analysis the sequence.

RESULTSThe first step of hearing screening of 460 newborn babies showed " refer" on the left ear of nine babies and on the right ear of three babies. Seven showed "refer" on bilateral with the the total of babies 19. After 42 days, they accepted the second step for hearing screening. 16 of the 19 were showed "pass" with OAE and AABR. One baby showed "pass" on the left ear, "refer" on the right ear with the OAE detection but bilateral "pass" with AABR. Two babies failed to accept the re-examination. The newborn gene screening showed five of the 460 babies had the positive response on the A1555G restriction enzyme assay. Of the five babies, one was proved to be the 12SrRNA A1555G mutation and three were the C1556T mutations and one sequence was normal. For the SLC26A4 gene screening, five were the heterozygote of IVS7-2A > G mutation were found and one was carrier the polymorphism of IVS7-18T > G and another was IVS6-62_63insGT heterozygote carrier. For the GJB2 gene screening, eight were 235delC heterozygote carriers, four were G109A heterozygote carriers. All the gene screening found 23 newborn babies of the 460 harbored the changes in the three genes. Of those, one was the 12SrRNA A1555G. pathogenic mutation and 13 were pathogenic heterozygote carriers, nine were the polymorphisms. It was worth to pay more attentions that A1555G mutation was found in the baby whose hearing screening was "pass" in the hearing screening as well as the 13 heterozygote carrier for GJB2 and SLC26A4 gene.

CONCLUSIONSIt might be one of the powerful strategy for adding the concept of newborn gene screening into the hearing screening for the purpose of early diagnosis and discovery the prelingual or late-onset or the high risk as well as the pathogenic carriers. On the basis of the research progress, it was necessary to develop the national newborn gene screening into the process of newborn hearing screening.

Connexins ; Evoked Potentials, Auditory, Brain Stem ; Female ; Hearing Disorders ; diagnosis ; genetics ; prevention & control ; Hearing Tests ; Humans ; Infant, Newborn ; Male ; Neonatal Screening ; Point Mutation