1.Analysis of Dangguibuxue decoction on rabbit schistosomiasis hepatic fibrosis through histopathological evaluation
Chinese Journal of Endemiology 2010;29(3):292-294
Objective To evaluate the treatment effects on schistosomiasis hepatic fibrosis with Dangguibuxue decoction.Methods Thirty-four rabbits weighing 2.0-2.5 kg were selected.The procedure for the model of schistosomiasis liver fibrosis was as follow.Thirty-four snails infected by schistosoma japonicum were placed in triangle flask to escape cercariain.Abdominal skin of each rabbit wag infected by 100 schistosoma japonicum cercariaes.All rabbits were housed in conventional cages at room temperature.After 13 weeks schistosoma japonicum infected,the rabbits'livers were examined by B-ultrasound examination every week.After 18 weeks of infection,B-ultrasound showed images of schistosomiasis liver fibrosis,the rabbits were thus identified as schistosomiasis liver fibrosis.Praziquantel Was as insecticide to treat rabbits.All rabbits were divided into treatment and control groups according to body weigh.The high and low dosage group were treated with Dangguibuxue decoction at 50 g/kg and 5 g/kg(n=11),respectively,and the control group with no treatment(n=12).Two groups were treated with Dangguibuxue decoction every day for 10 weeks.After 10 weeks of treatment,the rabbits were killed and the livers were obtained to go HE staining and histopathological examination under microscope.Results After 18 weeks of infection,B-ultrasound showed that the liver parenchyma Was coarse and net-shaped.having hyperechoie spots with lightbrand and facula.The liver's shape Was abnormal,portal and splenic vein diameter was widened.Histopathological examination showed that the high dosage group presented fibrosis,which was formed by collagen fiber extending to peripherM regions,low dosage group showed mild fibrosis,mild collagen fiber present with extension without compartment formation, while the control group presented severe fibrosis and severe collagen fiber present with a thickening of the partial compartments and the distribution of eggs nodules. The liver histopathological ranking results from different groups showed that the difference was significant (χ2= 13.585,P < 0.01), the difference between the treatment groups of high and low dosage and control group was significant,respectively(all P < 0.01 ). Conclusions The histopathology results showed that both high and low dosage Dangguibuxue decoction can reverse or eliminate schistosomiasis liver fibrosis in rabbits.
2.The mitochondrial and structural protein changes in dexamethasone-induced mouse thymocyte apoptosis
Tong WANG ; Yaoying ZENG ; Jingxian ZHAO ; Yi LIN ; Peiyan LIANG
Chinese Journal of Pathophysiology 2000;0(07):-
AIM: To study mitochondrial mass and structural protein changes in dexamethasone (DEX)-mediated mouse thymocyte apoptosis process. METHODS: DEX-induced mouse thymocyte apoptosis model was established. Annexin V-FITC/PI double staining was used to identify apoptotic and necrotic cells by flowcytometry, JC-1 staining was adopted to test mitochondrial membrane potential (△?_m), and cellular structural protein changes were studied with CFDA-SE staining. RESULTS: By 1?10~(-6) mol/L DEX stimulation, the apoptotic rate was 51.25%?5.51% and had significantly difference from control group (12.03%?2.00%); the necrotic rate in DEX group was 30.25%?3.67% and also had significantly difference from control group (10.11%?1.11%, P
3.Micron traditional Chinese medicine functional lining material:activating blood circulation and promoting fracture healing
Qing ZENG ; Guozhi HUANG ; Donghui LIANG ; Hongzhu DENG ; Yankui YI
Chinese Journal of Tissue Engineering Research 2016;20(38):5678-5683
BACKGROUND:As a new self-synthetized medical orthopedic fixation material, micron traditional Chinese medicine functional lining material can promote the secretion of alkaline phosphatase and cal us formation, decrease serum calcium level, as wel as increase serum phosphorus level and calcium-phosphorus product. OBJECTIVE:To further verify the therapeutic effect of micron traditional Chinese medicine functional lining material on fracture healing. METHODS:Twenty-four New Zealand rabbits were selected to prepare models of right radial fracture, and then were randomly al ocated into four groups:rabbits in micron group were treated with micron traditional Chinese medicine functional lining material (micron Panax notoginseng powder mixed with active ingredients of Daval ia divaricata and Carthamus tinctoriousplus) plus plaster external fixation;those in traditional Chinese medicine control group treated with ordinary Chinese medicine functional material plus plaster external fixation;those in lining material control group treated with ordinary lining material (sterile degreasing cotton) plus plaster external fixation;those in model control group received no treatment. The whole blood viscosity at high/middle/low shear rates and plasma viscosity were detected at 2, 4 and 6 weeks before and after modeling. Additional y, rabbits in each group were kil ed at 6 weeks after modeling, and radial samples were removed for histological observation. RESULTS AND CONCLUSION:The whole blood viscosity at high/middle/low shear rates and plasma viscosity in the micron group were al lower than those in the other three groups in the same period, and had significant differences from those in the model control group at 4 and 6 weeks after modeling (P<0.05 or 0.01). A large number of osteoblasts could be found, woven bone began to form and the bone marrow cavity was recanalized in the micron group;in the traditional Chinese medicine group, woven bone, chondrocytes and endochondral ossification coexisted and there was a connection between cal uses inside and outside of the broken end, but the bone marrow cavity was not recanalized. In contrast, neither connection between cal uses inside and outside of the broken end, nor recanalized bone marrow cavity appeared in the lining material and model control groups. In conclusion, micron traditional Chinese medicine functional lining material can promote fracture healing by improving microcirculation and accelerating osteoblast proliferation.
4.Determination of Gentiopicroside in Urine by LC/MS/MS
Yi FENG ; Jing SUN ; Yuanhui DENG ; Xing ZENG ; Weixiong LIANG
Traditional Chinese Drug Research & Clinical Pharmacology 2000;0(06):-
Objective To establish a HPLC/MS/MS method for the determinati on of gentiopicroside in human urine.Methods The urine sample was treated by solid-phase extraction with internal standard of caffeine;The RESCEK C8 colum n(150mm?2.1mm,5 ?m) was used as the analytical column with a mobilep hase consisting of methanol-10mmol? L-1 NH4AC buffer(pH=6.5)-acetonit rile(50∶40∶10,V/V),the flow rate was 0.2 mL? min-1;A triple quadruple tandem mass spectrometer was used as the detector,Electrospray ioniza tion source was applied and operated in positiveion mode.Gentiopicroside and caffeine were detected by monitoring the ion transition of m/z 374.1→ 195.2 and m/z 195.2→ 138.2 respectively.Results The linear range was 30~ 9000 ng?mL-1(r=0.9980) for gentiopicroside in human urine.The recovery was 91.10% ~ 9 6.21 %,The absolute recovery was 100.52% ~ 103.83%,The within-day and between-day precisions were less than 10 %.Conclusion The method is proved to be sensitive,accurate,rapid,specific.
5.Preparation and specificity of antibodies against ORF65 capsid protein of Kaposi′s sarcoma associated herpesvirus
Yuanhong CHEN ; Lijuan ZHAO ; Youlong LIANG ; Zhuohua ZHANG ; Yi ZENG
Chongqing Medicine 2015;(8):1091-1093,1096
Objective To prepare a high‐titer rabbit specific serum antibody against Kaposi′s sarcoma associated herpesvirus (KSHV) ORF65 capsid protein and identify the specificity of serum antibody .Methods Artificial synthetic peptide of ORF65 pro‐tein was emulsified with Freund adjuvant .4 rabbits were immunized with the prepared antigen by subcutaneous injection at various sites of skin of back and jaw once every two weeks .Immunization was carried out in total 4 times .The serum of the immunized rab‐bits was collected at a week after the last immunization .The titer of rabbit anti‐serum was assayed by ELISA .Specificity of the rab‐bit anti‐serum was analyzed by immunofluorescence assay and Western blot .Results The immunized rabbits produced high‐titer se‐rum antibody after total immunization .The highest titer of anti‐serum against ORF65 protein peptide was 1∶12 800 .The results of Immunofluorescence assay showed that antibody was binded in plasma of BCBL‐1 cell mostly ,which was consistent with the expres‐sion location of ORF65 in BCBL‐1 cell .Subsequently ,the data of Western blot revealed a specific band about 21 kD which accorded with the size of ORF65 protein .Meanwhile ,the expression of ORF65 in TPA treated BCBL‐1 cells was higher than the control cells ,which was consistent with the expression characteristics of lytic protein .Conclusion High‐titer specific rabbit serum antibody against KSHV capsid ORF65 antigen could be successfully prepared by rabbits immunization with ORF65 protein peptide .The pre‐pared antibody could be revealed immune reaction specificity with KSHV ORF65 protein .
6.Proteomics Research of Chinese Medical Syndromes of Steroid-induced Osteonecrosis of the Femoral Head
Ping ZENG ; Jingchao LIANG ; Yi ZHOU ; Gang QIN ; Tianxiao PAN
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(5):785-789,795
Objective To screen the differentially expressed proteins of blood stasis blocking tendon and vessel syndrome and liver-kidney deficiency syndrome of the steroid-induced osteonecrosis of the femoral head ( SONFH) by proteomic technology, so as to supply evidence for Chinese medical syndrome classification. Methods The serum was taken separately from 10 patients with blood stasis blocking tendon and vessel syndrome, 10 patients with liver-kidney deficiency syndrome and 10 healthy volunteers. The two dimensional electrophoresis combined with mass spectrometric method was applied to screen and identify the differentially expressed proteins, and then the obtained protein candidates were verified by Western blotting method. Results Seven proteins were identified from differentially expressed protein spots, including hemoglobin subunit delta, actin, complement C4, antithrombin-Ⅲ, apolipoprotein A-IV, leucine-rich alpha-2 glycoprotein and serum amyloid A-2 protein. We found that antithrombin-Ⅲ and serum amyloid A-2 protein had specific expression in the SONFH patients with blood stasis blocking tendon and vessel, and complement C4 and leucine-rich alpha-2-glycoprotein had specific expression in the SONFH patients with liver-kidney deficiency syndrome. The results of Western blot method showed that the expression levels of complement C4 and antithrombin-Ⅲ were down-regulated in SONFH patients ( P<0.05 compared with the healthy volunteers) , the down-regulation of complement C4 was more obvious in SONFH patients with liver-kidney deficiency syndrome (P<0.05) and the down-regulation of antithrombin-Ⅲ was more obvious in the patients with blood stasis blocking tendon and vessel (P<0.05), the results being accorded with those of proteomic detection. Conclusion Antithrombin-Ⅲand serum amyloid A-2 protein may be the specific serum protein markers of SONFH patients with blood stasis blocking tendon and vessel syndrome, and complement C4 and leucine-rich alpha-2-glycoprotein may be the specific serum protein markers of SONFH patients with liver-kidney deficiency syndrome.
7.Growth and developmental parameters and behavioral characteristics of infant rhesus monkeys
Zhigang LIANG ; Mingling WU ; Licai ZENG ; Zunwei YAO ; Sirui YI ; Rong LI ; Dong CAO ; Fengjun BI ; Li GONG ; Wen ZENG
Chinese Journal of Comparative Medicine 2016;26(4):24-34
Objective To explore the growth and developmental parameters and behavioral characteristics of rhesus monkeys during the first year of birth and to establish the background data.Methods A total of 18 (♂=11,♀=7 ) infant rhesus monkeys born from individually caged mothers and with known genetic background and postnatal days were monitored monthly for body weight, body height, head circumference, chest circumference, forelimb length, hind limb length, crown-rump length, tail length and anal-genital distance from postnatal day ( PND) 1 to 360, while hematology, blood chemistry and lymphocyte subsets were examined on PND 28, 175 and 360, and finger maze test was carried out on PND 208.Results The body weight showed linear growth with no significant difference between genders (P>0.05). Except for the anal-genital distance of male infants was significantly greater than that of female infants ( P<0.01 ) , no significant differences were observed between sexes in other morphological parameters.No significant differences of hematology were seen between genders (P >0.05).Compared with that at PND28, TP and BUN were significantly increased (P<0.01) while ALP decreased with no significant difference (P>0.05) at PND 175 and 360.Compared with that at PND28, CD4 +and CD4 +/CD8 +were significantly decreased ( P<0.01) while CD8+significantly increased ( P<0.01) at PND175 and 360.The number of sessions to solve task 2 in learning test was significantly greater than other tasks with females significantly less than males ( P<0.05) .The females had higher correct rate than males in the 2-day random memory test (P<0.05).Conclusions Body weight and morphological parameters show a linear growth.The PND.The RBC, HGB, LYMPH, TP, BUN, ALP, CD4 +, CD8 +and CD4 +/CD8+in hematology, blood chemistry and lymphocyte subsets show relevant changes to the growth and development of organs and systems in infants, which should be highly concerned in drug evaluation.The finger maze test indicates that female infants have better reversal learning and long-term memory than male infants.Background data and behavioral characteristics of infant rhesus monkeys during the first 12 months of birth are established in this study, which provide useful reference and support the evaluation of developmental and reproductive toxicity of drugs in rhesus monkeys.
8.Correlation between eIF3a and HE4 expression and ovarian cancer
Jing WANG ; Chenhui LUO ; Ying WANG ; Yuxi TANG ; Kaining FANG ; Liang ZENG ; Yi ZHANG
Journal of Central South University(Medical Sciences) 2014;(12):1240-1245
Objective: To investigate the correlation between eukaryotic translation initiation factor 3, subunit A (eIF3a) and human epididymis protein 4 (HE4) expression and ovarian cancer. Methods: RT-PCR or immunohistochemistry was used to examine eIF3a and HE4 mRNA or protein expression in ovarian tissues from patients with ovarian cancer (n=181) or benign ovariantumors, or from the healthy women. Results: hTere were signiifcant differences in mRNA and protein expression of eIF3a and HE4 among normal ovarian tissues, benign ovarian tumor tissues, and ovarian cancer tissues (P<0.05). hTere were signiifcant differences in mRNA expression of eIF3a and HE4 between the normal tissues and the ovarian cancer tissues, or between the benign ovarian tumor tissues and the normal tissues (P<0.001). hTe mRNA expression of eIF3a in the normal ovarian tissues was signiifcantly higher than that in the benign ovarian tumor tissues or that in the ovarian cancer tissues. hTe mRNA expression of HE4 was gradually increased from the normal ovarian tissues, the benign ovarian tumor tissues to the ovarian cancer tissues. hTe mRNA expression of HE4 in the ovarian cancer tissues was signiifcantly higher than that in the benign ovarian tumor tissues (P<0.001). Positive expression rates for eIF3a or HE4 protein in normal, benign tumor, and cancer tissues were 0, 66.7%, and 81.0% or 0, 27.8%, and 56.2%, respectively. hTere were signiifcant differences in positive expression rates of eIF3a protein and HE4 protein between the ovarian tumor tissues and benign ovarian tumor tissues, between the ovarian cancer tissues and the normal ovarian tissues, or between the benign ovarian tumor tissues and the normal ovarian tissues (P<0.001). hTe eIF3a protein expression was positively correlated with HE4 protein expression (r=0.575,P<0.05). Conclusion: The expressions of eIF3a and HE4 are associated with ovarian cancer, and extracellular regulated protein kinases may play a role in the interaction between eIF3a and HE4.
9.The application of non-invasive prenatal genetic sequencing for fetal chromosomal aneuploidy
Huinan WENG ; Jiaying LIANG ; Weihong ZENG ; Huixia TANG ; Yi SUN ; Jiangjun MA
International Journal of Laboratory Medicine 2015;(16):2386-2388
Objective To explore the efficiency and the clinical application value of non-invasive prenatal genetic testing for fetal chromosomal aneuploidy.Methods A total of 1 865 pregnant women treated in Guangdong Women and Children Hospital from January 201 1 to January 2013 were selected.Inclusion criteria:advanced age,prenatal screening for high risk,and fetal abnormality indicated by color ultrasonography,agreeing with non-invasive prenatal genetic testing.After non-invasive prenatal genetic testing, the pregnant women with positive result underwent cell culture and chromosomal karyotyping.Following the situations after deliv-ery were designed as the final criteria for definite diagnosis of fetal chromosomal aneuploidy.Results A total of 1 865 pregnant women underwent non-invasive prenatal genetic testing,of which 21 pregnant women were found with positive result,including 14 pregnant women with trisomy 21,5 pregnant women with trisomy 18,2 pregnant women with trisomy 13.The results of chromo-somal karyotyping after amniocentesis or umbilical cord blood puncture were designed as golden standard.Among the women with trisomy 21,one woman refused the prenatal diagnosis,self induced labor and could not be confirmed karyotype.No false positive case was found among the women with trisomy 18 and 13.No missed diagnosis was found among the pregnant women with negative result during follow-up after delivery.Through statistical analysis of non-invasive prenatal fetal genetic testing,the sensitivity for the trisomy 21 was 100%,and the accuracy was 92.9%.The sensitivity and accuracy for the trisomy 18 and 13 were 100%.Conclu-sion Non-invasive prenatal genetic testing can improve the diagnostic efficacy before delivery,reduce the birth of ill infants,and it is a quick,safe,easy-accepted and reliable prenatal diagnostic method,which is worthy to be popularized and an inexorable trend of development in the future.
10.Anatomic study of poking reduction and bone grafting technique for Hill-Sachs lesions
Yi HAO ; Xuan-liang RU ; Zeng-hui JIANG ; Hang LIN ; Jian HE ; Chun LIu ;
Chinese Journal of Trauma 2011;27(5):456-459
Objective To determine whether the poking reduction and bone grafting technique with guide through bony tunnel can correct a Hill-Sachs lesion. Methods A total of 30 cadaveric humeri were equally divided into three groups, 10 cadaveric humeri per group. Hill-Sachs lesions were replicated with a osseous defect involving 10% (group A ) , 20% (group B ) and 30% (group C ) of the articular surface. All the bone defects in each group were measured and the poking reduction and bone grafting technique with guide through a bony tunnel was performed in group B and group C. The preoperative and postoperative transverse arc length, longitudinal are length, depth and volume of the osseous defects in group B and group C were compared by using paired t test. Results Before reduction, the transverse arc length of the bone defects was ( 10.9 ± 1.4 )mm in group B and ( 16.3 ± 2.3 ) mm in group C ; longitudinal arc length was ( 22.4 ± 2.4 ) mm in group B and ( 28.0 ± 2.2 ) mm in group C ;depth was (6.9±0.9) mm in group B and (11. 1 ±0.9) mm in group C; volume was (708.7±93.9) mm3 in group B and (1338.3 ± 185.6) mm3 in group C. After reduction, the transverse arc length of the bone defects was (5.1 ± 2.4 ) mm in group B and ( 7.6 ± 3.6 ) mm in group C ; longitudinal arc lengthwas (10.5 ±4.9) mm in group B and (12.3 ±5.3) mm in group C; depth was (0.3±0.1 ) mm in group B and (0.4 ±0.1 ) mm in group C; volume was (48.9 ± 16.1 )mm3 in group B and (70.3 ± 37.9) mm3 in group C. The comparison of all the parameters showed statistical difference (P <0. 01 ). Conclusion The poking reduction and bone grafting technique with guide through a bony tunnel can effectively correct the Hill-Sachs lesions with humeral head osseous defects involving 20% -30% of the articular surface.