The dynamic changes of germination percentage, germination potential, thousand-seed weight, antioxidase activity in Desmodium styracifolium seeds with different storage time were tested, and electrical conductivity, contents of soluble sugar, soluble protein, starch in seed leach liquor were also determined in order to reveal the mechanism of seed deterioration. The results as the following. (1) The germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds declined, while the seed coat color darkened with the extension of storage time. (2) The activities of superoxide dismutase (SOD) and peroxidase (POD) decreased with the prolongation of storage period. The SOD activity declined fastest in 1,095-1,185 d of storage, while the POD activity declined significantly in 365-395 d of storage. (3) The electrical conductivity and the contents of soluble sugar, starch in seed leach liquor increased, while the content of soluble protein declined with the extension of storage time. (4) Correlation analysis indicated that the germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds have a significantly positive correlation with SOD and POD activity, while have a significantly negative correlation with the electrical conductivity, contents of soluble sugar and starch. It can be concluded that during the storage of D. styracifolium seeds, physiological and biochemical changes including decrease in antioxidase activity, rise in electrical conductivity, degradation effluent of soluble sugar and starch, degradation of soluble protein were the main factors leading to the seed deterioration.
Color ; Fabaceae ; chemistry ; enzymology ; growth & development ; metabolism ; Germination ; Peroxidases ; metabolism ; Plant Proteins ; metabolism ; Seeds ; chemistry ; enzymology ; growth & development ; metabolism ; Starch ; metabolism ; Superoxide Dismutase ; metabolism ; Time Factors

OBJECTIVETo study the impact of the water extract from Codonopsis thalictrifolia Wall (CTW) on the reproductive

METHODSWe divided 32 male SD infant rats into four groups of equal number to be treated intragastrical-system of male infant rats. ly with distilled water (control) and CTW at 10 g/kg (low dose) , 20 g/kg (medium dose), and 40 g/kg (high dose), respectively, twice a day for 2 weeks. Then we killed the rats, measured the levels of testosterone (T), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the serum, obtained the testis weight, body weight, testis visceral coefficient and sperm concentration, and detected sperm viability, sperm motility and the level of cyclic adenosine monophosphate (cAMP) in the Leydig cells, followed by

RESULTSCompared with the control group, the low-dose, me-analysis of differences among different groups using the SPSS software. Medium-dose and high-dose CTW groups showed significant decreases in the serum T level ([3.09 +/-0.42] vs [1.22 +/-0. 32] , [1.06 +/- 0.29] and [0.57 +/-0.18] nmol/L, P<0.01), testis weight ([1.40 +/-0.16] vs [0.96 +/-0.09], [0.92 +/-0.11] and [0.91 +/- 0.08] g, P <0.01), and sperm concentration ([1.03 +/-0.16] vs [0.19 +/-0.07], [0.17 +/-0.08] and [0.16 +/-0.07] x 10(6)/ml, P <0.01), but a dramatic elevation in the testis visceral coefficient ([42.22 +/- 3.02] vs [51.39 +/- 3.09], [52.28 +/- 4.86] and [54.13 +/-6.06] mg/10 g, P <0.01); the medium- and high-dose CTW groups exhibited remarkable increases in the levels of serum LH ([13.62+/-0.89] vs [14.69 +/-0.12] and [14.93 +/-0.28] ng/L, P<0.01) and FSH ([4.32 +/-0.18] vs [4.77 +/-0.23] and [4.89 +/-0. 38] IU/L, P <0.05); all the three CTW groups showed markedly inhibited serum T secretion ([1.85 +/- 0.18] vs [1.42 +/-0.15], [1.12+/-0.18] and [0.88 +/-0.21] nmol/L, P<0.01) and intracellular cAMP ([5.51 +/-0.12] vs [4.39+/-0.06], [4.28 +/-0.07] and [4.11 +/- 0.10] nmol/L, P <0.01) in the Leydig cells.

CONCLUSIONThe water extract from CTW may reduce the synthesis of testosterone in the serum of male infant rats through the PKA pathway and consequently inhibit their testicular development and sperm production and affect the development of their reproductive system.

Animals ; Codonopsis ; chemistry ; Cyclic AMP ; metabolism ; Leydig Cells ; metabolism ; Male ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Testosterone ; blood ; Urogenital System ; drug effects

OBJECTIVEIn oder to treat periodontitis by using tissue engineering and gene engineering technology, the article established an transient expression system of bone marrow stromal cells (BMSC) modified by osteoprotegerin (OPG) gene and detected its expression using eukaryotic secreted expression pSecTag2/B-OPG plasmid.

METHODSBy solation and culture of BMSC in vitro, the identified recombined plasmid was transiently transfected into BMSC by Lipofectamine 2000 and OPG expression in BMSC was determined by RT-PCR and Western blot in 6 weeks.

RESULTSThe fragments of the recombinant plasmid digested with Hind III, EcoR I and BamH I and examined by 10 g/L agarose electrophoresis, were consistent with predicted size. The sequence of OPG was identical to the sequence provided by GeneBank [gi:33878056]. OPG transcribing in BMSC was confirmed by RT-PCR and OPG sustainable expressing in BMSC was detected by Western blot in 39 days.

CONCLUSIONThe transiently expression system of BMSC modified by OPG gene was successfully established.

Humans ; Mesenchymal Stromal Cells ; Osteoprotegerin ; Tissue Engineering ; Transfection

Objective To investigate the clinical effect of Zhuyun Ning Recipe Ⅱ in treating spleen-kidney deficiency type of threatened abortion with polycystic ovarian syndrome(PCOS). Methods A total of 40 qualified outpatients and inpatients admitted in the First Affiliated Hospital of Heilongjiang University of Chinese Medicine were included into the study. Twenty patients diagnosed as spleen-kidney deficiency type of threatened abortion with PCOS complicated with insulin resistance (IR) were included into group A, 20 patients with spleen-kidney deficiency type of PCOS non-IR threatened abortion were included into group B,and 20 normal women with early pregnancy were included into group C. Groups A and B were given oral use of Zhuyun Ning RecipeⅡfor 30 days, and group C did not receive any medication. The scores of traditional Chinese medicine (TCM) symptoms, blood beta-human chorionic gonadotropin (β-HCG), serum progesterone, homeostatic model assessment for insulin resistance index(HOMA-IR), interleukin-18(IL-18), and interleukin-20(IL-20) of groups A and B were observed before and after treatment,and were compared to those of group C. Results (1)After treatment for 30 days,the observation indexes of group A were much improved(P < 0.05 or P < 0.01 compared with those before treatment), and the indexes in group B except for HOMA-IR were also improved (P < 0.05 or P < 0.01). The inter-group comparison showed that the differences of blood β-HCG, serum progesterone,and IL-10 were insignificant (P > 0.05);HOMA-IR and IL-18 levels in group A were higher than those in groups B and C, and the differences were significant (P < 0.01). However, the differences of HOMA-IR and IL-18 were insignificant between group B and group C (P > 0.05).(2)During the treatment, neither adverse reaction nor toxic or side effect was found.(3)The follow-up results showed that 5 cases of group A,2 cases of group B,and 4 cases of group C had full-term normal delivery,and the health state of newborn babies was quite well. One case of group A had pregnant diabetes at late pregnant stage,and the other patients of group A got through the middle and late pregnancy,without any pregnant complications. Conclusion Zhuyun Ning Recipe Ⅱ is effective on relieving symptoms of threatened abortion patients with PCOS,and can improve the related laboratory indexes and pregnancy outcomes of the patients.

OBJECTIVETo analyze the superiorities of acupoint catgut-embedding therapy, discuss its law of clinical application and provide scientific decision-making for clinical treatment.

METHODSLiteratures on acupoint catgut-embedding therapy in the recent 40 years were selected, input, examined and verified, picked up and analyzed by establishing database with the modern computer technology.

RESULTS(1) One thousand and seventy-five literatures were input. It shows that the acupoint catgut-embedding therapy has an extensive application in all departments, especially in the internal department, accounting for 48.54% (50/103) of the total disease category. It has the most extensive application on treatment of epigastric pain, with the frequency of 102 times, and obesity of 74 times. The next is surgery, accounting for 14.56% (15/103). The major application is on low back pain and leg pain with the frequency of 79 times. Psoriasis, with the frequency of 30 times, holds the major application in dermatological department. And blepharoplasty, with the frequency of 30 times, gains the most application in department of ophthalmology and otorhinolaryngology. (2) In the included literatures, selection of adjacent acupoints and distal acupoints are held as the major method of acupoint selection. The adjusted lumbar puncture needle is taken as the major tool for the acupoint catgut-embedding therapy. And catguts of different sizes are adopted for the operation. (3) Analysis of the therapeutic effect shows that acupoint catgut-embedding therapy has obvious effect in all departments, especially in surgery and dermatology, with the total effective rate over 90%.

CONCLUSIONEpigastric pain, obesity, epilepsy, asthma, abdominal pain, facial paralysis and constipation of the internal medicine, low back pain and leg pain of the surgical department, psoriasis of the dermatological department and blepharoplasty of the department of ophthalmology and otorhinolaryngology are considered as the dominant diseases for acupoint catgut-embedding therapy.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Catgut ; utilization ; Facial Paralysis ; therapy ; Humans ; Low Back Pain ; therapy ; Obesity ; therapy

OBJECTIVETo examine the expression of molar root patterning gene 1 (Mrp1) and predict the Mrp1 structure by bioinformatics analysis.

METHODSA pair of Mrp1-specific PCR primers were designed, and RT-PCR method was used to study the mRNA's expression pattern in rat molar root and other organs. Gene positioning and other protein sequence prediction were carried out by chromosome analysis and other bioinformatics analysis.

RESULTSMrp1 was expressed not only in the molar but also in the developing pancreas, liver, lung and kidney tissues. Mrp1 was located in the 18q12.3 chromosome of the rats and the Mrp1 amino acids sequence had about 37% homology with a known protein Uroplakin IIIb (p35) which was an urothelial differentiation membrane molecular marker. A trans-membrane structure, 5 PKC phosphorylation sites and 4 CKII phosphorylation sites in Mrp1 were found.

CONCLUSIONSMrp1 has a broad expression in different developing organs, and it may have a important function in the rat tooth root development.

Animals ; Computational Biology ; Gene Expression Regulation, Developmental ; Genes ; Molar ; growth & development ; Molecular Sequence Data ; Protein Sorting Signals ; genetics ; Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Tooth Root

OBJECTIVETo evaluate the effect of SMU.2055 gene on acid resistance of Streptococcus mutans.

METHODSA SMU.2055-dificient mutant strain of S. mutans was constructed using homologous recombination technique. The growth of the wild-type and mutant strains was monitored in both normal and acidic conditions. The lethal pH level, glycolysis, proton permeability, cell permeability and biofilm formation of the two strains were compared.

RESULTSPCR and sequence analyses verified the successful construction of the SMU.2055-dificient mutant strain. The growth and biofilm formation capacity of the mutant strain were obviously lowered in both normal and acidic conditions. The mutant strain also showed increased lethal pH level, proton permeability, and cell permeability with impaired H-ATPase activity in acidic conditions, but its minimum glycolytic pH remained unaffected.

CONCLUSIONThe SMU.2055-deficient S. mutans mutant exhibits a lowered acid resistance, which affects the growth, lethal pH, proton permeability, H-ATPase activity, cell permeability and biofilm formation but not the minimum glycolytic pH of the mutant strain.

OBJECTIVETo study the effect of Shugan Shuru Granule (SSG) on the p53 gene expression in patients with hyperplastic disease of breast (HDB) to indirectly explore the mechanism of SSG's effect on HDB on the molecular pathological level.

METHODSSixty-six patients with HDB were allocated in the treated group and the control group, with the former treated with SSG and the latter not. All patients underwent breast operation and their diseased mammary tissues were cut out, sectioned, and observed under microscopy with HE staining and immunohistochemical staining, with ABC method adopted to estimate the degree of hyperplasia and p53 gene expression. The severity of HDB was classified into normal, mild, moderate and severe grades (marked as 0 to III), according to the degree of hyperplasia in the mammary gland.

RESULTSHyperplasia in the control group mostly belonged to grade I-III before treatment, showing overgrowth of gland and proliferation of glandular epithelial cells, which were high columnar shaped, more stratified, with papillary or substantive dysplasia. While in the treated group, most belonged to grade 0-I after SSG treatment, with proliferated gland and dysplasia recovered to normal or disappeared. The positive rate of p53 gene expression in the treated group was 9.09%, and in the control group 39.39%, comparison between the two groups showing significant difference (P < 0.01), the intensity in the former was significantly weaker than that in the latter.

CONCLUSIONSSG could not only inhibit the proliferation of mammary duct epithelia and lobuli, but also inhibit the over-expression of P53. Therefore, it could be regarded as an effective remedy for treatment of HDB and prevention of mammary cancer genesis.

Adult ; Breast Diseases ; drug therapy ; pathology ; surgery ; Breast Neoplasms ; prevention & control ; Drugs, Chinese Herbal ; administration & dosage ; Epithelium ; pathology ; physiology ; Female ; Gene Expression ; drug effects ; Humans ; Hyperplasia ; Mammary Glands, Human ; pathology ; physiology ; Middle Aged ; Tumor Suppressor Protein p53 ; genetics

BACKGROUNDApoptosis is involved in the adaptive responses of bone to mechanical loading. The purpose of this study was to investigate the effects of tensile forces on osteoblast apoptosis and the related mechanism by analyzing the expression of caspases, Bcl-2, and Bax.

METHODSPrimary osteoblasts were harvested from neonatal rat calvaria and were subjected to cyclic tensile forces for 72 hours using Flexcell 4000 strain unit in Minimum Essential Medium (MEM) with 10% fetal calf serum (FCS) or with serum deprivation. Apoptosis was tested by flow cytometry using annexin V/PI staining. Caspase-3 activity was analyzed via Elisa. The gene expression of caspase-8, -9, Bcl-2, and Bax was quantified by reverse transcription (RT)-PCR.

RESULTSIn 10% FCS condition, no significant difference in cell apoptosis was found between the stretched and non-stretched osteoblast cultures. Serum withdrawal resulted in higher apoptosis rate in the osteoblasts with increased caspase-3 activity, and elevated expression of caspase-9 and Bax. Six-percent elongation of stretch attenuated the cell apoptosis induced by serum starvation, concurrent with a decrease in caspase-3 activity, a decline of caspase-8 expression, and an elevation of Bcl-2 level. On the contrary, 12% elongation of stretch increased caspase-3 activity and promoted the apoptosis with an elevated expression of caspase-8 and Bax. No significant change of caspase-9 expression was identified upon force application.

CONCLUSIONSThese results suggested that tensile forces regulate cell apoptosis of primary rat osteoblasts through caspase-3 and caspase-8 signaling cascade. Light forces rescue the cells from serum deprivation-induced apoptosis by elevating Bcl-2 expression, while heavy forces promote the apoptotic insult by inducing Bax expression.

Animals ; Apoptosis ; physiology ; Caspase 8 ; genetics ; metabolism ; Caspase 9 ; genetics ; metabolism ; Caspases ; genetics ; metabolism ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Osteoblasts ; cytology ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; bcl-2-Associated X Protein ; genetics ; metabolism

AIM:To investigate the effect of high-mobility group box-1 (HMGB1) expression knockdown on the invasion ability of breast cancer cells induced by tumor necrosis factor-α (TNF-α).METHODS:HMGB1 siRNA was used to transfect into the breast cancer MDA-MB-231 cells.The expression of HMGB1 at mRNA and protein levels was determined by RT-qPCR and Western blot.After the MDA-MB-231 cells with HMGB1 expression knockdown were treated with TNF-α, the apoptosis rate was analyzed by flow cytometry, the cell invasion ability was measured by Transwell assay, and the cell migration ability was detected by cell scratch test.The protein expression of E-cadherin, MMP-2, N-cadherin, MMP-9 and Bax was determined by Western blot.RESULTS:The expression of HMGB1 at mRNA and protein levels in the MDA-MB-231 cells transfected with HMGB1 siRNA was significantly lower than that in the non-transfected cells (P<0.05).The apoptosis rate in the cells was increased after TNF-αtreatment, and the cell invasion and migration abilities were also increased.The protein level of E-cadherin in the cells was decreased, the protein level of N-cadherin was increased, and the protein levels of MMP-2, MMP-9 and Bax were also increased (P<0.05).After the MDA-MB-231 cells with HMGB1 expression knockdown were induced by TNF-α, the apoptotic rate was increased, the invasion and migration abilities were decreased, the protein levels of E-cadherin and Bax were increased, and the protein levels of N-cadherin, MMP-2 and MMP-9 were decreased, as compared with the cells only induced by TNF-αwithout knockdown of HMGB1 expression (P<0.05).CONCLUSION:Knockdown of HMGB1 expression enhances the apoptosis of breast cancer cells induced by TNF-α, and inhibited the cell invasion, migration and epithelial-mesenchymal transition induced by TNF-α.The mechanism may be related with the changes of protein expression of MMP-2, MMP-9 and Bax.