1.Effect of catalpol on activity of osteoblasts/osteoclasts and osteoblast ERα/βmRNA expression in osteoblast-osteoclast co-culture system
Manxiang LAI ; Li YANG ; Ronghua ZHANG
Chinese Journal of Pathophysiology 2015;(7):1242-1246
[ ABSTRACT ] AIM: To investigate the effect of catalpol on the activity of osteoblasts ( OB ) and osteoclasts ( OC) , and OB estrogen receptor ( ER) α/βmRNA expression in the OB-OC co-culture system.METHODS: OB and OC were isolated from the SD rats of 1 and 5 days old.In the OB-OC co-culture system, different concentrations of catalpol including low dosage (0.05 , 0.1, 0.5 and 1 mg/L), middle dosage (2, 5 and 10 mg/L), and high dosage (20, 50 and 100 mg/L) were added into the culture medium to detect the changes of OB proliferation by MTT assay.The catalpol at maximal dosage was added to OB section to detect the alkaline phosphatase ( ALP) activity of OB by pNPP method.The mRNA expression of ERα/βin the OB treated with catalpol in the co-culture system was detected by RT-PCR.The catalpol at maximal dosage was added to OC group to detect the activity of OC by microscopy and tartrate-resistantacid phosphatase ( TRAP) activity detection.RESULTS:In 0.05~2 mg/L catalpol groups, the proliferation of OB was significantly in-creased as compared with control group in the co-culture system, and it reached the maximum value when catalpol was at 0.05 mg/L, while in 5~100 mg/L catalpol groups, the proliferation of OB was not increased.The ALP activity of OB in 0.05 mg/L catalpol group was higher than that in control group.The catalpal at 0.05 mg/L promoted the mRNA expression of ERβin OB in the co-culture system, but did not increase the mRNA expression of ERαas compared with control group. Catalpol at 0.05 mg/L obviously inhibited the bone resorption and the TRAP activity in OC.CONCLUSION: Catalpol stimulates the proliferation and activity of OB, inhibits the bone resorption and activity of OC, and increases the mRNA ex-pression of ERβin OB in the OB-OC co-culture system, suggesting that high mRNA expression of ERβmay be the regula-tory pathway of catalpol in response to bone metabolism.
2.Common bacterias distribution and antibiotics analysis of wounds secretion in surgery
Lihua ZHANG ; Xiaobing ZHANG ; Zhigang LAI ; Li ZHANG
International Journal of Surgery 2010;37(5):319-322
Objective To investigate the common Pathogenic bacterias distribution and antibiotics analysis of wounds secretion in surgery so as to provide scientific evidence for antibiotics choice of wound infection in clinical. Methods Antimicrobial susceptibility test was carried out by Kirby-Bauer method. Results was analyzed according to CLSI 2009. The data were analyzed using WHONET 5.4 software. Results Totally 483 strains of pathogens were isolated from 628 specimens, which isolating rate was 76.9%. The most bacterial was E. coil(21.2% ), followed by S. aureus(18.0%), P. aeruginosa(9.7% ), K. pneumoniae(6.4% )and A. baumannii(6.2% ). Gram bacterias were all sensitive to imipenem. 30. 6% of S. aureus were identified as methicillin-resistant Staphylococcus, 86.6% of coagulase negative Staphylococcus isolates were methilillin-resistant. Conclusions The bacteria species is diversity in wound secretion , and their drug resistance is serious, so medical staff should to pay attention to the bacterial culture of clinical specimen and the monitoring bacteria resistance and anti-bacteria drug should be managed by system to control bacterial infection and drug resistance.
3.Application of MR-cine in evaluation on functional dyspepsia
Mingyu ZHANG ; Jincheng LI ; Ying LAI ; Kailiang CHENG ; Mengchao ZHANG
Journal of Jilin University(Medicine Edition) 2016;42(2):385-389
Objective:To explore the value of MR-cine for assessment of the duodenum peristalsis in the patients with functional dyspepsia.Methods:25 patients with functional dyspepsia were selected according to the diagnostic criteria of RomeⅢ of functional gastrointestinal disorders (FGIDs)as case group and 25 cases of healthy volunteers who had been screened out in clinic were used as normal control group. After fasting for 8 h,supine position was performed.1.5TGEHDxMR was used to scan the coronal,axial and oblique coronal fast steady state precession (FIESTA)sequence of gastric and duodenal descending part.The images of 5 and 15 min after drinking 600 mL mannitol solution (concentration 2.5%)were collected and sent to MR AW4.4 workstation.The duodenal motility in the patients with functional dyspepsia was measured by recording the times of duodenum and measuring and calculating the percentage occlusion of duodenum contractions (PDC). Results:After drinking mannitol, the oblique coronal scan of all subj ects clearly displayed the anatomical structure and peristaltic wave of descending part of duodenum. After drinking 5 and 1 5 min of mannitol,the peristaltic wave frequencies in case group were lower than those in normal control group. The descending duodenum PDC of the patients in case group was significantly lower than that in normal control group (P<0.01)5 min after drinking. The PDC in normal control group 15 min after drinking was lower than 5 min (P<0.05);the duodenal PDC 15 min after drinking of mannitol had no statistical difference between case group and normal control group (P>0.05).Conclusion:MR-cine can evaluate preliminarly the duodenum peristalsis of the patients with functional dyspepsia. The diagnosis of FGIDs can be further studied by using the noninvasive MR-cine examination technique.
4.Diagnosis and Treatment of acute subdural hematoma
Mingwen ZHANG ; Runlong LAI ; Yong LI ; Tianbo ZHANG ; Yu CHEN
Chinese Journal of Primary Medicine and Pharmacy 2011;18(2):150-152
Objective To investigate the clinical characteristics and therapeutic strategy of acute subdural hematoma. Methods A restrospective study was carried out with a total of 94 consecutive ASDH patients who were confirmed through computed tomographic scan and obtained the clinical characteristics by experienced neurosurgeons.15 cases werepure acute subdural hematoma and the other 79 cases were acute compound subdural hematoma according to CT scan. Results In accordance with the GOS,36 cases had good recovery,19 cases had moderate disability,17 cases had severe disability and 22 cases dead. Conclusion The most important treatment for pure acute subdural hematom was to diagnose the source of bleeding, and acute compound subdural hematoma had poor prognosis than pure acute subdural hematoma since the traumatic severity. Early decraniuim by large bone flap to treat acute compound subdural hematoma could improve survival rate,reduce the fatality rate and decrease postoperative complications.
5.The effects of bifidobacterial secretory adhesin on the stress response of intestinal epithelial cells
Shishun ZHONG ; Zhenshu ZHANG ; Shumei LI ; Zhuosheng LAI ; Dongliang LI
Chinese Journal of Digestion 2011;31(11):744-749
ObjectiveTo observe the effects of bifidobacterial sectory adhesin on NF-κB DNA binding activity,cytoplasm IκB,nuclei NF-κBp65,TNF-α,IL-1β,and IL-8 expression in intestinal epithelial cell post stress response.MethodsThe cultured intestinal epithelial cell line Lovo cells were divided into 5 groups,which included directly stimulated by lypopolysaccharide (LPS) (100 ng/ml) or H2O2 (200 μmol/L) for 3 hours groups,bifidobacterial sectory adhesin (30μg/ml) pre-incubation for 30 minutes and then treated by LPS (100 ng/ml) or H2O2 (200 μmol/L) for 3 hours groups and control group with no treatment.NF-κB DNA binding activity was evaluated by electrophoretic mobility shift assay (EMSA).The cytoplasm IκB and nuclei NF-κBp65 expression were determined by Western blot.The expressions of TNF-α,IL-1β and IL-8 at mRNA level were detected by semi-quantatitive assay of RT-PCR.ResultsThe expressions of NF-κB DNA binding activity [(6.20±0.35) times and (4.16 ± 0.52) times of that of non treated control group,respectively] and nuclei NF-κBp65 [ (0.64±0.05) and (0.67±0.06)] were higher in cells after LPS or H2()2 stimulated,however the cytoplasm IκB expression [ (0.28 ± 0.10) and (0.39 ± 0.12) respectively] was weaker.After bifidobacterial sectory adhesin pre-incubation,the expressions of NF-κB DNA binding activity and nuclei NF-κBp65decreased obviously,but cytoplasm IκB expression increased.After treated with LPS or H2O2,the mRNA expressions of TNF-α,IL-1β and IL-8 were significantly increased [LPS treated group2 (0.92±0.10)、(0.38±0.03)、(1.44±0.25),H2O2 treated group:(0.89±0.13)、(0.36±0.06)、(1.42±0.18)].After bifidobacterial sectory adhesin pre-incubation,their expressions decreased.The mRNA expressions of TNF-α,IL-1β and IL-8 have significantly positive correlation with NF-κB DNA binding activity.ConclusionsThere is a significant inhibition role of bifidobacterial sectory adhesin in LPS and H2O2 induced NF-κB DNA binding activity in intestinal epithelia cells.The activation of NFκB may associate with the regulation of TNF-α,IL-1β and IL-8 proinflammatory cytokines expression.
6.REPORT OF HLA DISTRIBUTION IN 2315 VOLUNTEER DONORS OF CHINESE BONE MARROW BANK FROM NORTHWEST CHINA
Tianbo JIN ; Ya GAO ; Hongbo ZHANG ; Junhai ZHAO ; Jianghua LAI ; Shuping LAI ; Shengbin LI
Journal of Pharmaceutical Analysis 2005;17(1):45-48
Objective To report the HLA data of volunteer donors of Chinese bank from Northwest China and characterize the distribution of HLA genes in Northwest China. Methods HLA-A, B antigens of 2315 volunteer donors were examined by the method of microlymphocytetoxicity (MLT) test .The antigen frequencies(AF) were assessed by directly counting; and based on that gene frequencies(GF) were calculated. HLA data from other population were collected and distribution characteristics were compared. With the raw data, Hardy-Weinberg equilibrium, statistical parameters of forensic medicine interest for HLA were computed. Results A total of 18 specific antigens were detected in HLA-A and the most frequent antigen was A2 . AF and GF were 0.5136 and 0.3026, respectively. A total of 42 specific antigens were detected in HLA-B and the most frequent antigen was A13. Its AF and GF were 0.1978 and 0.1044, respectively. The heterozygosity(H), polymorphism information content(PIC), discrimination power(DP) and probability of paternity exclusion (PPE) of HLA-A were 0.8215, 0.8212, 0.9356 and 0.7798 accordingly; while H,PIC, DP and PPE of HLA-B were 0.9322, 0.9322, 0.9878 and 0.9528. Conclusion The polymorphism of HLA-A,B genes is characteristic in Chinese. In this research, the genetic trait of HLA in 2315 volunteers is consistent with Northern Han population.
7.Three-stage induced membrane technique combined with anterior and posterior double-plate fixation for a total talus defect after infection
Hongning ZHANG ; Guodong SHEN ; Yunxuan ZOU ; Xue LI ; Kangyong YANG ; Zhibin LAI ; Junhui LAI ; Yongzhan ZHU
Chinese Journal of Orthopaedic Trauma 2021;23(5):401-408
Objective:To evaluate three-stage induced membrane technique combined with anterior and posterior double-plate fixation in the treatment of a total talus defect after infection.Methods:Included in this study were 11 patients with talus infection who had been treated at Department of Orthopaedics, Foshan Hospital of Traditional Chinese Medicine from January 2014 to December 2018. They were 8 males and 3 females, aged from 23 to 63 years (mean, 37.0 years). The infection followed re-implantation after open dislocation of total talus in 4 cases, internal fixation for open talus fracture of Gustilo type Ⅲa in 3 cases and surgery of open ankle fracture of Gustilo type Ⅲc in 2 cases, and was complicated with ankle intraarticular tuberculosis in 2 cases. The three-stage operations consisted of debridement, total talus resection, implantation of antibiotic bone cement and vacuum sealing drainage at the first stage, change of bone cement, re-debridement, wound closure or flap covering at the second stage 7 to 10 days later, and reconstruction after infection control using anterior and posterior double-plate fixation and induced membrane technique at the third stage 6 to 12 weeks later. Assessment of lower limb shortening was performed by comparing the full length of the leg between the normal and affected sides; the functions were assessed by comparing the ankle-hindfoot scores of American Orthopedic Foot and Ankle Society (AOFAS) and visual analogue scale (VAS) between preoperation and the final follow-up.Results:The 11 patients were followed up for an average of 24.3 months (from 12.2 to 37.5 months). Superficial skin necrosis was observed in 2 patients and injury to superficial peroneal nerve in one. Absolute calcification of the autograft area was observed in all patients, leading to ankle fusion. The final follow-ups observed no significant difference in the full length of the leg between the normal and affected sides [(380.4±35.5) mm versus (376.3±32.8) mm] ( P>0.05) , a significant increase in the ankle-hindfoot AOFAS scores from preoperative 28.0±3.4 to 72.8±5.4, and a significant decrease in VAS scores from preoperative 5(5,6) to 0(0,1) (all P<0.05). Slight varus developed in 2 patients and slight ankle stiffness in 3; recurrence of infection or breakage of implants was found in none of the patients. Conclusion:Three-stage induced membrane technique combined with anterior and posterior double-plate fixation can effectively control infection of the talus, maintain the length and reconstruct the function of the lower limb after a total talus defect.
8.Analysis and state of HCV genotype 6a infection
Rui ZHANG ; Junqiang LI ; Lijun LIU ; Shaocai DU ; Lai WEI
Journal of Peking University(Health Sciences) 2003;0(06):-
Objective: To investigate the infection state of hepatitis C virus genotype 6a in China.Methods: Three(95,126,150)HCV genotype 6a serum samples were identified by digesting 5′NCR with compound enzyme method.Then,HCV 5′NCR and NS5B fragments were amplified from these samples by RT-PCR assay and sequenced.The phylogenetic trees of the samples were analyzed and compared with 24 HCV complete gene sequences from GenBank.Results: The sequencing reports on 5′NCR showed "CA" bases in 3 serum samples(95,126,150) were inserted into-145 site,and the sequences of 3 serum samples had the highest homology with sequence Y12083(0.934,0.930,and 0.926,respectively).The results of the phylogenetic trees suggested these 3 serum samples belonged to HCV genotype 6a.The sequencing reports on NS5B showed the 3 serum samples also had the highest homology with HC-J4(0.934,0.930,and 0.926,respectively),and the results of the phylogenetic trees suggested these 3 serum samples belonged to HCV genotype 1b.To exclude the influence of amplification efficiency of primers,NS5B fragments were amplified by HCV genotype 6a specific primers and no amplification products appeared.Conclusion: There are different results of HCV genotype by analyzing 5′NCR and NS5B in 3 samples infected with HCV genotype 6a.It may be related with gene recombination.It suggests HCV genotype should be analyzed on more than two regions.
9.Utilization of Uracil-DNA glycosylase for combining reverse transcription and anti-contamination with polymerase chain reaction in hepatitis C virus
Shaocai DU ; Rui ZHANG ; Junqiang LI ; Lai WEI
Journal of Peking University(Health Sciences) 2003;0(04):-
Objective: To develop a hepatitis C virus(HCV) reverse transcription-polymerase chain reaction (RT-PCR) assay using Uracil-DNA glycosylase (UDG) for amplicon contamination control and evaluate the temperature and UDG concentrations for anti-contamination. Methods: In this new HCV RT-PCR assay, reverse transcription, UDG anti-contamination and the first PCR were carried out at the same time. The layer candles were used to prevent the contamination in the second PCR. dU-DNA was used as quality control for UDG anti-contamination and templates to determine the sensitivity of the new HCV RT-PCR assay. HCV cDNA was detected by DNA enzyme immunoassay (DNA-EIA). Results: Complete degradation of amplicon DNA was observed on the conditions of 0.2 u UDG per reaction volume respectively at 37 ℃ and 42 ℃ for 40 min. The anti-contamination condition also could eliminate all detectible dU-DNA, including the highest concentration of amplicon DNA.The 1∶104 dilution of the HCV RNA sample containing 2.110?105copies/mL copies of RNA could be detected. Conclusion: Our results indicate that this new RT-PCR assay can control the contamination stringently and is sensitive as well.
10.Intraarterial embolization combined with resection for the treatment of huge tumors in the buttocks
Biming LIU ; Yu ZHAO ; Hua ZHANG ; Yongqiang LI ; Lai WEI
Chinese Journal of General Surgery 2009;24(4):304-306
Objective To evaluate a combination therapy for huge tumors in the buttocks.Methods A total of 11 patients from our hospital were collected,among them 5 cases were of hemangioma,4 cases of neurofibroma,2 cases of soft tissue sarcoma.Before definite surgical resection all cases received tumor embolization with silk thread and gelatin-sponge article using Seldinger's technic.Subsequently,all patients underwent a successful tumor resection. Results Superselective embolization for all the cases'feeding arteries resulted in recession of the tumors and relatively well-demarcated margins,and all the lumps became softer.and the local pain was alleviated.Surgical resection could be radical with avoidance of fatal intraoperative hemorrhage.The 5 cases of hemangioma had a average operative bleeding of 450 ml,4 cases of neurofibroma had 420 ml,2 cases of soft tissue sarcoma had 150 ml.No patients needed intraoperative and postoperative blood transfusion.The operation time was about 2-3 hours,the normal tissues were preserved and the contour and function of the diseased limbs were very good.One case had a delayed incision healing,and the others had a healing by the first intention.There was no recurrence and other complications (like deep venous thrombosis)during a follow-up period of 4-8 months. Conclusions Surgical resection combined with interventional embolism for the treatment of huge tumors in the buttocks can reduce the risk of bleeding effectively during operation.It can improve the success rate of operation leading to satisfactory results.