2.Influence of Human Chorionic Gonadotropin on Testicular Germ Cell Apoptosis of Bilateral Cryptorchidism in Rats
kai-sheng, LV ; yan-fang, REN ; cui-ping, SONG
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To investigate the influence of human chorionic gonadotropin on testicular germ cell apoptosis of bilateral cryptorchidism in rats.Methods Immature male rats (22 day-old Sprague Dawley) were subjected to bilateral cryptorchidism and injected with 20 U human chorionic gonadotropin from the 22th day to the 34th day every other day. Sham operation rats were as control group. At age 35 days and 60 days, rats were sacrificed for detection germ cell apoptosis by terminal- deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL). Results There were significant difference of apoptosis index (AI) between bilateral cryptorchidism and sham operation groups(P0.05).Conclusion AI of cryptorchidism increases and HCG addes the number of apoptotic germ cells.
3.Effects of Radix Paeoniae Alba Extract on Distribution and Expression of 5-HT3A/3B R in Frontal Lobe of Premenstrual Syndrome Rat Model with Liver-qi Depression Pattern
Fang LI ; Chunhong SONG ; Sheng WEI ; Huiyun ZHANG ; Mingqi QIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(11):2267-2271
This study was aimed to elucidate the 5-HT3R molecular mechanism of Radix Paeoniae Albaextract (RPAE) as drug intervention in the treatment of premenstrual syndrome (PMS) model rats with liver-qi depression pattern.PMS model rats with liver-qi depression pattern were prepared.And then,the model was treated with RPAE.The protein distribution of 5-HT3AR and 5-HT3BR in the frontal lobe was evaluated by the immune fluorescence technology and western blot.The results showed that there were positive expressions of 5-HT3AR and 5-HT3BR in frontal lobe of rats in each group.Compared with the normal group,the 5-HT3AR and 5-HT3BR protein expression levels of the frontal lobe in the model group increased significantly (P < 0.05).Compared with the model group,the 5-HT3AR protein expression level in the frontal lobe decreased significantly after RPAE treatment (P < 0.05).In conclusion,RPAE regulated the protein expressions of 5-HT3AR and 5-HT3BR in frontal lobe,which may be one of the mechanisms for its treatment of PMS with liver-qi depression pattern.
4.Preparation technology and dissolution research of Blumea volatile oil suppository.
Song WANG ; Yong-Heng ZHAO ; Yi-Sheng ZHOU ; Fang-Fang LI
China Journal of Chinese Materia Medica 2014;39(10):1805-1810
OBJECTIVETo research the preparation technology and dissolution of Blumea volatile oil suppository.
METHODIn order to establish the content determination and methodology inspection method of Blumea volatile oil plug, the extraction process of Blumea volatile oil was optimized by using orthogonal test. Optimization on the investigation to the suppository matrix by melting time, appearance and dissolution was carried on. The best prescription craft was determined by determining the best molding temperature, dosage of the matrix and complementary makings. The determination method of dissolution was established by investigating different dissolution method and its impact on the preparation of dissolution.
RESULTThe best conditions of steam distillation extracted Blumea volatile oil was as followed, the ratio of gardenia to liquor 1:6, 2.5% drug amount of sodium, 8 hours of extracting time. The optimum temperature for mold was 60-65 degrees C. Preparation technique of Blumea volatile oil suppository was stable, which after 45 minutes and 3 h in pH 4.5 PBS released at least 70% and 90%.
CONCLUSIONBlumea volatile oil suppository with rational prescription, simple preparation and good stability.
Asteraceae ; chemistry ; Chemistry, Pharmaceutical ; methods ; Distillation ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Oils, Volatile ; chemistry ; isolation & purification ; Plant Oils ; chemistry ; isolation & purification ; Solubility ; Temperature
5.THE EFFECT OF APP17 ON MODULATION OF Bcl-2,Bax,CREB,AKt,AIF EXPRESSIONS IN HIPPOCAMPAL NEURONS OF D-GALACTOSE INDUCED BRAIN AGING OF THE MICE
Pengwen WANG ; Zhiwei ZHAO ; Lina SONG ; Fang YANG ; Zhijuan JI ; Shul SHENG
Acta Anatomica Sinica 1955;0(03):-
Objective To investigate the effect of the peptide APP17 on regulating the expression of Bcl\|2,Bax,cAMP response element binding Protein(CREB),Ser\|Thr kinase B/protein kinase B(Akt/PKB),apoptosis inducing factor(AIF) in neurons of the hippocampus from the D\|gal mouse. Methods D\|gal mouse models were established by injection of D\|gal.In experimental group,these models were injected with APP17 petide subcutaneously and their brain sections were taken after 3 months of survival.The immunohistochemical staining of these sections was then performed with Bcl\|2,Bax,CREB,Akt,AIF antibody. Results Bax,AIF positive neurons were widely distributed in the hippocampus of the D\|gal mice,and the cytoplasm was darkly stained.In contrast,positive cells in the hippocampus were poorly stained in those normal mice and the APP17 peptide\|treated D\|gal mice.But Bcl\|2,CREB,AKt positive neurons were widely distributed in the hippocampus of those normal mice and the APP17 peptide\|treated D\|gal mice,and the cytoplasm was darkly stained.In contrast,positive cells in the hippocampaus were poorly stained in the D\|gal mice.Conclusion\ The expression of Bax and AIF could be increased in the hippocampus of D\|gal mice.But the expression of Bcl\|2,CREB,AKt decreased in the hippocampus of D\|gal mice.The APP17 can regulate the distribution of Bcl\|2,Bax,CREB,Akt,AIF in the brain of D\|gal mice and return them to normal situation.\;[
6.Effect of Radix Bupleuri Extract on 5-HT3R Pathway in Primarily Cultured Rat Hippocampal Neurons
Fang LI ; Jieqiong WANG ; Chunhong SONG ; Sheng WEI ; Huiyun ZHANG ; Mingqi QIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(11):2224-2228
This study was aimed to investigate the effect of Radix Bupleuri extract (RBE) on 5-HT3R channel currents of primarily cultured hippocampal neurons in depression emotion rats.Depression emotion model ratswere duplicated.RBE was used for drug intervention.And then,the rats were evaluated by the open-field test (OFT) and the sucrose preference test.Serum of rats in each group was collected and then added into the primary cultured hippocampal neurons for 24 h.The 5-HT3R channel currents were recorded by the whole-cell patch clamp.The results showed that compared with the normal group,the total score of OFT in the model group was significantly decreased (P < 0.01); the sucrose preference ratio decreased obviously (P < 0.01); and the current density value of primary cultured hippocampal neurons in serum of the model group was significantly higher (P <0.01).Compared with the model group,the total scores of OFT in the RBE group and fluoxetine group increased significantly (P < 0.05,P < 0.01); the sucrose preference ratio also increased obviously (P < 0.05,P < 0.05); and current density value of the primary cultured hippocampal neurons in serum of the RBE group and fluoxetine group decreased significantly (P < 0.01,P < 0.01).It was concluded that RBE can effectively correct the abnormal 5-HT3R channel currents of rats with depression emotion,which may be one of the central mechanisms in the treatment of depression emotion.
7.Effects of serum enatninine Gumibao (Chinese character: see text) on the aroliferation and differentiation of osteoblast induced by dexamethasone.
Yi SONG ; Hong JIAN ; Dao-fang DING ; Ling-hui LI ; Guo-qing DU ; Jin-tao LIU ; Hong-sheng ZHAN
China Journal of Orthopaedics and Traumatology 2014;27(8):668-672
OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.
METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.
RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.
CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.
Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley
8.Etablishment of cartilage degeneration model by IL-1 beta in vitro.
Dao-fang DING ; Jian PANG ; Yi SONG ; Guo-qing DU ; Yue-long CAO ; Hong-sheng ZHAN ; Yu-xin ZHENG
China Journal of Orthopaedics and Traumatology 2015;28(7):648-653
OBJECTIVETo establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.
METHODSThe femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde, decalcified, dehydrated, embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type II collagen, MMP13, Sox9 and ADAMTS5 were analyzed by immunofluorescence.
RESULTSBoth the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group, which indicated that cartilage became degenerated. The expression levels of both type H collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.
CONCLUSIONThe model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro.
Animals ; Cartilage Diseases ; genetics ; metabolism ; Collagen Type II ; genetics ; metabolism ; Disease Models, Animal ; Femur Head ; metabolism ; Humans ; In Vitro Techniques ; Interleukin-1beta ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; genetics ; metabolism
9.Low-dose radiation induces endoplasmic reticulum stress and activates PERK-CHOP signaling pathway in mouse testicular cells.
Fang FANG ; Ping-Sheng GONG ; Xiang-Fu SONG ; Shou-Liang GONG ; Zhi-Cheng WANG
National Journal of Andrology 2012;18(9):777-782
OBJECTIVETo explore the correlation of low-dose radiation with endoplasmic reticulum stress and the activation of the PERK-CHOP signaling pathway in mouse testicular cells.
METHODSHealthy Kunming mice were randomly assigned to time-effect (0, 3, 6, 12 and 24 h of irradiation at 75 mGy) and dose-effect (12 h of irradiation at 0, 50, 75, 100 and 200 mGy) groups. The contents of H202 and MDA were measured by colorimetry with the agent kits, the expressions of GRP78, PERK and CHOP mRNA detected by quantitative RT-PCR, and the levels of GRP7B, PERK, phosphorylated PERK (pho-PERK) and CHOP proteins determined by Western blotting and image analysis.
RESULTSAfter whole-body irradiation of the mice with 75 mGy, the content of H2 02 in the testis tissue was increased with time prolongation, while that of MDA decreased slightly at 3 and 6 h and then increased with the lengthening of time, both increased significantly at 12 and 24 h as compared with those at 0 h (P < 0. 05, P < 0. 01). Apart from reduced levels of GRP78 mRNA at 3 and 24 h and GRP78 protein at 6 h after irradiation, significant increases were found in the mRNA expressions of GRP78 at 12 h, PERK at 3,6, 12 and 24 hand CHOP at 12 and 24 h (P < 0.05, P < 0.01), as well as in the protein levels of GRP78 at 12 and 24 h, pho-PERK at 3, 12 and 24 h and CHOP at 3, 6, 12 and 24 h in comparison with those at 0 h (P < 0. 05, P < 0. 01). No obvious regularity was observed in the change of the PERK protein expression. After 12 h of whole-body irradiation, the content of H202 was increased at 50, 75 and 100 mGy, but decreased slightly at 200 mGy, while that of MDA was increased with dose increasing, with significant increases in the content of H2 02 at 75 and 100 mCy and in that of MDA at 75, 100 and 200 mGy as compared with the 0 mGy group. Apart from the reduced levels of GRP78 mRNA at 50 and 200 mCy, significant increases were found in the mRNA expressions of PERK at 75, 100 and 200 mGy and CHOP at 50, 75, 100 and 200 (P c 0. 05, P < 0.01) as well as in the protein levels of GRP78 at 100 and 200 mGy, pho-PERK at 50, 100 and 200 mGy and CHOP at 50, 75, 100 and 200 mCy as compared with those at 0 mGy (P < 0. 05, P < 0. 01). There were differences in the changes of different protein expressions, but no obvious regularity was seen in the change of the PERK protein expression.
CONCLUSIONLow-dose radiation can induce endoplasmic reticulum stress in mouse testicular cells, and activate the PERK-CHOP signaling pathway.
Animals ; Endoplasmic Reticulum Stress ; radiation effects ; Heat-Shock Proteins ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Radiation Dosage ; Radiation, Ionizing ; Signal Transduction ; radiation effects ; Testis ; cytology ; metabolism ; radiation effects ; Transcription Factor CHOP ; metabolism ; Whole-Body Irradiation ; eIF-2 Kinase ; metabolism
10.Relationship between maximum body mass index and type 2 diabetes mellitus among adults in Heilongjiang province
Wen WEI ; Sheng-Yuan LIU ; Fang-Fang ZENG ; Song-Po YAO ; Hai-Tao ZHANG ; Gang WAN ; Min ZHONG ; Bin-You WANG
Chinese Journal of Epidemiology 2010;31(3):251-255
Objective To determine the prevalence and distribution of type 2 diabetes mellitus (T2DM) and the relationship between maximum body mass index (MAXBMI) and T2DM. Methods From June to August, 2005, a stratified cluster sampling of 1071 permanent residents in communities, over 20 years old, from 4 districts and 1 county of Mudanjiang was chosen. The prevalence of T2DM, and the association between T2DM and different levels of the MAXBMI, current BMI were studied. Results The prevalence in the communities was 7.09% and in those with past maximum BMI≥28 kg/m~2, it was 12.10%. With the increase of past MAXBMI levels, the risk of T2DM patients also increased significantly(trend X~2=17.387 23, P<0.0001). Data from multifactor analysis showed that MAXBMI in the past was positively related to T2DM (OR=3.06, P=0.0013). In T2DM patients, the group with MAXBMI≥27.4 kg/m~2 had higher 2-hour postprandial blood glucose than those with lower MAXBMI (P=0.0408). When compared with low maximum BMI group in normal blood glucose population, the group with higher MAXBMI (≥ 25.4 kg/m~2) had higher blood glucose and greater change of BMI. Conclusion In both groups that patients with T2DM and with normal glucose, in order to control blood glucose better, researchers should not only concern about the influence of the MAXBMI in the past, but also pay attention to constantly keep BMI at the normal range.