1.Experimental research of Ruangan granule for the treatment of liver fibrosis in rats
Kun NA ; Liang WU ; Ying LI ; Ying ZHENG ; Jingri XIE
International Journal of Traditional Chinese Medicine 2015;(4):339-344
Objective To observe theRuangan granule on liver fibrosis in rats liver pathology change, the influence of hepatic function and hepatic fibrosis indexes, and to discusses the mechanism of its action to provide the basis for clinical prevention and treatment of liver fibrosis.Methods A total of 105 Wistar rats were randomly divided into a normal control group, a model group and a colchicines group, and Dahuang-Zhechong pill group, high-, medium- and low-doseRuangan granule groups (n=15 in each group). Liver fibrosis was induced by carbon tetrachloride and a high-cholesterol diet. After modeling, the low-, medium- and high-doseRuangan granule groups were intragastric administratedRuangan granule mixed suspension 3.6, 7.2, 14.4 g/(kg?d), respectively;Dahuang-Zhechong pill group was administrated with Dahuang-Zhechong pellets mixed suspension of 0.18 g/(kg?d); the colchicine group was intragastric administrated with colchicine mixed suspension of 0.108 mg/(kg?d); and the normal control group and the model group were intragastric administrated with the equal volume of distilled water. All rats were intragastric administrated for 8 weeks. HE staining and Masson trichromatic collagen staining were used to observe the pathological changes of liver While the change of AST, ALT, PH, TP and serum HA, LN, C-Ⅳ, PCⅢin blood serum were detected. Results Masson trichromatic collagen staining showed that, the percentage of liver collagen fiber area in rats of theRuangan granule high-dose group was significantly decreased (7.06 ± 1.18) % compared with model group (23.49 ± 1.34) %, colchicine group (11.35 ± 1.83) %, rhubarb worm pill group (15.27 ± 1.22) %,Ruangan granule medium-dose group (14.52 ± 1.75) %, and low dose group (16.08 ± 1.56) % (P<0.05 orP<0.01). Compared with model group,Ruangan granule high-dose group rats serum AST (75.86 ± 5.23 U/Lvs. 157.62 ± 24.04) U/L, the ALT (80.15 ± 5.94 U/Lvs. 160.58 ± 26.47) U/L, PH (52.58 ± 4.98μg/Lvs. 98.66 ± 6.75)μg/L significantly reduced, TP (74.19 ± 3.56 g/Lvs. 51.73 ± 5.92)g/L increased significantly (P<0.01).Ruangan granule high-dose group rats serum HA (277.22 ± 106.34 ng/mlvs. 553.19 ± 172.38 ng/ml), LN (89.82 ± 5.68 ng/mlvs. 134.25 ± 10.64 ng/ml), C-Ⅳ (47.94 ± 8.65 ng/mlvs. 84.18 ± 13.83 ng/ml), PCⅢ (16.53 ± 4.88 ng/mlvs.31.57 ± 5.35 ng/ml) decreased significantly in the model group (P<0.01).ConclusionRuangan granule has obvious effects for resisting liver fibrosis.
2.Effects of Ruangan granule on transforming growth factor-β1/Smads signaling pathway in liver fibrosis in rats
Kun NA ; Liang WU ; Ying LI ; Ying ZHENG ; Jingri XIE
International Journal of Traditional Chinese Medicine 2015;(1):47-51
Objective To investigate the effects of Ruangan granule on transforming growth factor-β1(TGF-β1)/Smads signaling pathway in liver fibrosis in rats. Methods A total of 105 Wistar rats were randomly divided into normal control group, model group and colchicine, Dahuang-Zhechong pill group, high-, medium- and low-dose Ruangan granule groups (n=15 in each group). Liver fibrosis was induced by carbon tetrachloride and a high-cholesterol diet. After modeling, the low-, medium- and high-dose Ruangan granule groups were intragastric administrated Ruangan granule mixed suspension 3.6, 7.2, 14.4 g/(kg?d), respectively;Dahuang-Zhechong pill group was administrated with Dahuang-Zhechong pellets mixed suspension of 0.18 g/(kg?d);the colchicine group was intragastric administrated with colchicine mixed suspension of 0.108 mg/(kg?d);and the normal control group and the model group were intragastric administrated with the equal volume of distilled water. All rats were intragastric administrated for 8 weeks. The expressions of TGF-β1, Smad3 and Smad7 proteins in the liver tissue were detected with immunohistochemical staining method. The expressions of TGF-β1, Smad3, Smad7 mRNAs in the liver tussue were detected by RT-PCR. Results The expressions of TGF-β1 (2.59 ± 0.99 vs. 0.43 ± 0.21) and Smad3 (2.56 ± 0.67 vs. 0.41 ± 0.18) proteins and TGF-β1 mRNA (2.25 ± 0.21 vs. 0.71 ± 0.09) and Smad3 (2.34 ± 0.03 vs. 0.78 ± 0.12) mRNAs in the model group were significantly increased than those in the normal control group (all P<0.01). Compared with the model group, the expressions of TGF-β1 (1.12 ± 0.27 vs. 2.59 ± 0.99) and Smad3 (1.05 ± 0.34 vs. 2.56 ± 0.67) proteins in the high-dose Ruangan granule group decreased significantly, the expression of Smad7 increased significantly (2.33 ± 0.62 vs. 0.36 ± 0.18), and the expressions of TGF-β1 (1.09 ± 0.11 vs. 2.25 ± 0.21) and Smad3 (1.10 ± 0.02 vs. 2.34 ± 0.03) mRNAs decreased significantly, the expression of smad7 mRNA (1.18 ± 0.13 vs. 0.38 ± 0.11) increased significantly (P<0.05). Conclusions Ruangan granule can regulate the TGF-β1/Smads signaling pathway via down-regulation of TGF-β1, Smad3 and up-regulation of Smad7 in liver fibrosis in rats.
3.Effect of isorhynchophylline on head-shakes behavior and levels of monoamine neurotransmitter in model rats with Tourette syndrome
Yang YAO ; Kun LIU ; Yu YANG ; Minfan WU ; Yu LIANG
Chinese Journal of Behavioral Medicine and Brain Science 2016;25(1):29-33
Objective To explore the effect of rhynchophylline and isorhynchophylline on headshakes behavior and levels of monoamine neurotransmitter in model rats with Tourette syndrome.Methods 40 Wistar rats were randomly divided into DOI-induced head-shakes rats (HSR group),haloperidol group,rhynchophylline group and isorhynchophylline group with 10 in each group.The inhibitory effects of rhynchophylline and isorhynchophylline were estimated by observing the HSR behavior.Dopamine (DA) and 5-hydroxytryptamine(5-HT) in the rat striatum were detected by the enzyme-linked immunosorbent assay (ELISA) method.The 5-HT2A receptor mRNA expression in prefrontal lobe cortex of the rats was measured by real-time PCR.Results Compared with HSR group,the head shakes of the rats in haloperidol group and isorhynchophylline group were significantly decreased(P<0.01),and no change of head-shakes number was observed in rhynchophylline group (P>0.05).There was no significant difference of head-shakes number between the haloperidol group and isorhynchophylline group(P>0.05).Compared with HSR group,DA levels in the rat striatum were significantly decreased in isorhynchophylline group and haloperidol group((152.35± 5.80) μ~L vs (111.19±4.30) μg/L,(152.35±5.80) μg/L vs (126.42±3.17) μg/L,P<0.01),while DA levels in the rat striatum in rhynchophylline group were not changed ((152.35±5.80) μg/L vs (142.71±5.51) μg/L,P>0.05).There was no significant change of 5-HT2A receptor mRNA expression in rat prefrontal lobe cortex in every group(P>0.05).Conclusion Isorhynchophylline may have an inhibitory effect on rats with DOI-induced HSR.Isorhynchophylline may decrease the DA levels in the rat stratum with DOI-induced HSR.Rhynchophylline has no significant inhibitory effect on head-shakes behavior and DA levels in the rat stratum with DOI-induced HSR.
4.Mechanism of Shenci capsule combined with cisplatin in reversing DDP resistance of human lung adenocarcinoma via PI3K/AKT/mTOR signa-ling pathway
Liqun XU ; Ronghua ZHANG ; Ying ZOU ; Jingjie PAN ; Xiaodong WU ; Lijian YU ; Kun LIANG
Chinese Journal of Pathophysiology 2017;33(3):500-504
AIM:To investigate the effect of Shenci capsule combined with cisplatin ( DDP) in reversing DDP resistance by PI3K/AKT/mTOR signaling pathway in nude mice bearing A 549/DDP tumor.METHODS:The patient-de-rived lung adenocarcinoma A 549/DDP cell xenograft model was established .The tumor-bearing nude mice were randomly divided into control group , Shenci capsule group , DDP group and Shenci capsule combined with DDP group .The mice in control group was treated with normal saline , while the mice in other groups were treated with different drugs for 21 d.After treatment, the mice were killed and lung cancer tissues were collected .Flow cytometry was used to analyze the cell cycle and apoptosis.FQ-PCR was used to determined the mRNA levels of PTEN , P-glycoprotein, PI3K, AKT and mTOR in A549/DDP lung tumor .RESULTS:Compared with control group , the cell proliferation in all the drug treatment groups was inhibited .Compared with other drug treatment groups , Shenci capsule combined with DDP blocked the cell cycle of A 549/DDP cells at G2/M phase, promoted the apoptosis rate , increased the mRNA expression of PTEN and inhibited the mRNA expression of P-glycoprotein, PI3K, AKT and mTOR.CONCLUSION:Shenci capsule increases the sensitivity of A 549/DDP resistant cells in nude mice to DDP by blocking PI 3K/AKT/mTOR signaling pathway , increasing the expression of PTEN or inhibiting P-glycoprotein-mediated resistance pathway .
5.The Protamine-like DNA-binding Protein P6.9 Epigenetically Up-regulates Autographa californica Multiple Nucleopolyhedrovirus Gene Transcription in the Late Infection Phase
Ying PENG ; Kun LI ; Rongjuan PEI ; Chunchen WU ; Changyong LIANG ; Yun WANG ; Xinwen CHEN
Virologica Sinica 2012;27(1):57-68
Protamines are a group of highly basic proteins first discovered in spermatozoon that allow for denser packaging of DNA than histones and will result in down-regulation of gene transcription[1].It is well recognized that the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) encodes P6.9,a protamine-like protein that forms the viral subnucleosome through binding to the viral genome[29].Previous research demonstrates that P6.9 is essential for viral nucleocapsid assembly,while it has no influence on viral genome replication[31].In the present study,the role of P6.9 in viral gene transcription regulation is characterized.In contrast to protamines or other protamine-like proteins that usually down-regulate gene transcription,P6.9 appears to up-regulate viral gene transcription at 12-24 hours post infection (hpi),whereas it is non-essential for the basal level of viral gene transcription.Fluorescence microscopy reveals the P6.9's co-localization with DNA is temporally and spatially synchronized with P6.9's impact on viral gene transcription,indicating the P6.9-DNA association contributes to transcription regulation.Chromatin fractionation assay further reveals an unexpected co-existence of P6.9 and host RNA polymerase Ⅱ in the same transcriptionally active chromatin fraction at 24 hpi,which may probably contribute to viral gene transcription up-regulation in the late infection phase.
6.Levels of interleukin-6,interleukin-10 and T cell subsets in bronchoalveolar lavage fluid of children with refractory mycoplasma pneumonia
Kun LIANG ; Ling TIAN ; Xiaolei WU ; Guangju HUANG ; Xiuhua DAI ; Huiyu ZHANG
Chinese Pediatric Emergency Medicine 2014;21(11):689-692
Objective To explore the interleukin (IL)-6,IL-10 and T cell subsets levels in bronchoalveolar lavage fluid(BALF) of children with refractory mycoplasma pneumonia.Methods A total of 53 children with refractory mycoplasma pneumonia were selected as the observation group,30 children with bronchial foreign body in our hospital were chosen as controls during the same period.ABC-double antibody sandwich ELISA method was used to detect IL-6,IL-10 levels and the CD3 +,CD4 + and CD8 + T levels were measured by means of flow cytometry in BALF.Results The IL-6 and IL-10 levels in BALF of children in the observation group were (63.25 ± 18.61) ng/ml,(31.83 ± 8.33) ng/ml respectively,and they were significantly higher than those of the controls[(30.51 ± 1.34) ng/ml,(11.01 ± 2.91) ng/ml] (P < 0.05,respectively).The percentage of CD3 +,CD4 +,CD8 + T cells and the ratio of CD4 +/CD8 + T cells in BALF of the observation group were (48.47 ± 2.88)%,(21.16 ± 6.29)%,(23.04 ± 4.63)%,0.94 ± 0.33,respectively,and they were significantly lower than those of the controls [(64.24 ± 3.06) %,(34.34 ± 7.59) %,(26.71 ±5.29)%,1.56-±0.67] (P<0.05,respectively).Conclusion The IL-6,IL-10 levels in BALF of children with refractory mycoplasma pneumonia significantly increased,suggesting that cell-mediated immunity play an important role in the pathogenesis of refractory mycoplasma pneumonia.
7.Polyphosphate kinase 1-coding gene (ppk1) is involved in the oxidative stress resistance in uropathogenic Escherichia coli by modulating the expression of katG and katE genes
Jingyi OU ; Liang PENG ; Xuelian ZHU ; Kun LUO ; Jiayun PAN ; Xiaoman WU
Chinese Journal of Microbiology and Immunology 2017;37(4):263-268
Objective To investigate the role and the mechanism of ppk1 gene (coding for polyphosphate kinase 1) in oxidative stress resistance in uropathogenic Escherichia coli (UPEC).MethodsMutant strains with ppk1-deletion (△pk1) and complemented strains (△pk1-C) were constructed based on the UPEC strain CFT073.A comparative analysis was conducted to analyze survival rates of CFT073, △pk1 and △pk1-C strains at different time points while they were under oxidative stress.Differences in protein expression between CFT073 and △pk1 strains were analyzed using mass spectrometric analysis.Differences between CFT073 and △pk1 strains in expression of katG and katE genes were analyzed using real-time quantitative RT-PCR.Results The survival rate of △pk1 strains was lower than that of CFT073 strains at every time point, while the survival rate of △pk1-C strains was basically the same as that of CFT073 strains.Gel image analysis and mass spectrometric analysis revealed that six proteins were down-regulated and one was up-regulated in △pk1 strains as compared with those in CFT073 strains.Expression of the catalase-coding genes katG and katE in △pk1 strains were respectively (20.5±8.2)% and (20.9±6.9)% of those in CFT073 strains (P<0.05).Conclusion The ppk1 gene plays an important role in oxidative stress resistance in UPEC by modulating the expression of catalase-coding genes katG and katE.
8.Minimally invasive percutaneous osteosynthesis for the treatment of 53 patients with complex ankle fractures.
Chang-Kun LI ; Bin ZHANG ; Xian-Wu YANG ; Xiang CHENG ; Wei DAI ; Yun LIANG
China Journal of Orthopaedics and Traumatology 2014;27(2):157-160
OBJECTIVETo explore the surgical method and its clinical effects of minimally invasive osteosynthesis on the treatment of complex ankle fractures.
METHODSFrom January 2007 to December 2011, 53 patients with complex ankle fractures were treated with minimally invasive osteosynthesis. There were 31 males and 22 females, with an average age of 38.2 years old (ranged, 18 to 65). According to the system of Lauge-Hansen, 32 fractures were supination external rotation injury (grade WV), 13 fractures were pronation external rotation (grade III or IV), 5 fractures were pronation abduction (grade III); and 3 fractures can not be classified due to serious comminution fracture of fibula. According to the system of Denis-Weber, there were 4 cases with type A, 34 cases with type B and 15 cases with type C. Seven cases were open fractures. The duration from injuries to operation ranged from 2 hours to 14 days with an average of 5 days. The sequence of reduction and fixation of ankle fractures was firstly posterior malleolus, then medial malleolus and lateral malleolus, and inferior tibiofibular syndesmosis lastly. The fractures of posterior malleolus were reduced and fixed through anterior ankle approaches; the fractures of medial and lateral malleolus were percutaneously fixed with bolts or blade plate or tensile force band; and inferior tibiofibular syndesmosis were firmly fixed if necessary. Baird-Jackson scoring system was used to evaluate clinical effects.
RESULTSForty-eight patients were followed up from 10 to 36 months with an average of 13 months. The fractures got healing with an average time of 12 weeks (ranged, 10 to 18). According to the Baird-Jackson scoring system, the mean score of ankle function was 94.7 +/- 4.2, and 28 cases obtained excellent results, 15 good, 3 fair and 2 poor. One case experienced superficial infections and was cured by changing dressings, 2 cases experienced fixed syndesmosis screw breakage.
CONCLUSIONThe surgical method of minimally invasive osteosynthesis can ensure the anatomical join restoration, protect the blood supply of fracture end, rebuild the function of ankle joint, obtain satisfactory clinical results in treating complex ankle fractures.
Adolescent ; Adult ; Aged ; Ankle Fractures ; Ankle Joint ; physiopathology ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods
9.Adult bilateral Crowe type-IV developmental dysplasia of the hip:is the prosthesis an optimal therapeutic strategy?
Xiaoliang SUN ; Liang WU ; Guofeng WU ; Jian ZHOU ; Kun WANG ; Jianwei ZHANG ; Liping ZHOU ; Zongming GONG ; Zhenyu TANG
Chinese Journal of Tissue Engineering Research 2014;(40):6408-6412
BACKGROUND:Crowe type-IV developmental dysplasia of the hip presented complete dislocation of the femoral head, significantly shortening lower limb, serious adverse development of upper segment of acetabulum and femur, and even deformity. At present, there are no reports concerning its therapeutic strategy.
OBJECTIVE:To explore the therapeutic strategy of adult bilateral Crowe type-IV developmental dysplasia of the hip.
METHODA total of 12 cases of adult bilateral Crowe type-IV developmental dysplasia of the hip, who were treated in the Department of Orthopedics, The First People’s Hospital of Changzhou from January 2000 to January 2013, were col ected. There were 5 males and 7 females, at the age of 19-47 years old, averagely 33.5 years old. After two or three weeks of skeletal traction, they received bilateral total hip arthroplasty. Hip rotation center was reconstituted at the level of the“true”acetabulum. The subtrochanteric osteotomy was performed for patients with reduction difficulty. For patients with femoral neck anteversion>40°, subtrochanteric rotary osteotomy was performed.
RESULTS AND CONCLUSION:Disappearance of hip pain, satisfied move function and normal gait were gained after operation. Lower limbs recovered to be isometric. The average leg lengthening was 3.1 cm (range, 2.5 to 4.8 cm). One patient affected sciatic nerve irritation. The average fol owed-up time was 3.5 years. No fracture, dislocation or loosening of the prosthesis appeared. The post-operative Harris Hip Score averaged 86.3. By reconstruction of the hip rotation center in the“true”acetabular level, correction of femoral neck anteversion, reconstruction of function of the abduction muscle and proper selection of prosthesis, total hip arthroplasty for adult bilateral Crowe type-IV developmental dysplasia of the hip could obtain good curative effects.
10.Prenatal diagnosis of prelingual deafness by determination of SLC26A4 gene mutation
Hao HU ; Lingqian WU ; Desheng LIANG ; Yong FENG ; Fang CAI ; Kun XIA ; Qian PAN ; Zhigao LONG ; Heping DAI ; Jiahui XIA
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
G mutation of SLC26A4, the parents and the second child were carriers of the same mutation, while the fetus had a wild-type form. Conclusion It is feasible to identify deafness related genes by screening for GJB2 and SLC26A4 mutation, thus providing correct prenatal diagnosis and avoiding deaf delivery of baby.