Objective:To evaluate the effect of emodin on liver injury in a mouse model of intestinal ischemia-reperfusion (I/R) and the role of heme oxygenase-1-mediated autophagy.Methods:Twenty-four SPF-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (Sham group), I/R group, emodin group (E group) and emodin plus HO-1 inhibitor Zinc Protoporphyrin Ⅸ (ZnPP) group (ES group). The intestinal I/R injury model was established by clamping the superior mesenteric artery for 45 min followed by 120 min of reperfusion. Emodin 40 mg/kg dissolved in 5% methylcellulose sodium was given by gastric gavage once a day for 5 days before ischemia in E group. Emodin 40 mg/kg dissolved in 5% methylcellulose sodium was given by gastric gavage once a day for 5 days before intestinal I/R, and ZnPP 7.5 mg/kg was injected via the tail vein at 12 h before ischemia in ES group. Orbital venous blood samples were collected at the end of reperfusion for determination of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations. Then the mice were sacrificed, and liver tissues were obtained for microscopic examination of the pathological changes (after HE staining) and for determination of the activity of superoxide dismutase (SOD), content of malondialdehyde (MDA), expression of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) mRNA (by fluorescent quantitative polymerase chain reaction), the expression of HO-1, autophagy-related protein Beclin1 and microtubule-associated protein 1 light chain 3 (LC3) (by Western blot). The LC3-Ⅱ/Ⅰ ratio was calculated. Results:Compared with Sham group, the activity of SOD was significantly decreased, the content of MDA and serum ALT and AST concentrations were increased, the expression of IL-6 and TNF-α mRNA and HO-1 was up-regulated, the expression of Beclin1 was down-regulated, the LC3-Ⅱ/Ⅰ ratio was decreased ( P<0.05), and the pathological changes of liver tissues were found in I/R group. Compared with I/R group, the activity of SOD was significantly increased, the content of MDA and serum ALT and AST concentrations were decreased, the expression of IL-6 and TNF-α mRNA was down-regulated, the expression of HO-1 and Beclin1 was up-regulated, the LC3-Ⅱ/Ⅰ ratio was increased ( P<0.05), and the pathological changes of liver tissues were significantly attenuated in E group ( P<0.05). Compared with E group, the activity of SOD was significantly decreased, the content of MDA and serum ALT and AST concentrations were increased, the expression of IL-6 and TNF-α mRNA was up-regulated, the expression of HO-1 and Beclin1 was down-regulated, the LC3-Ⅱ/Ⅰ ratio was decreased ( P<0.05), and the pathological changes of liver tissues were aggravated in ES group. Conclusions:Emodin can alleviate liver injury induced by intestinal I/R in mice, and the mechanism may be related to the activation of HO-1-mediated autophagy.

OBJECTIVE: To explore the technique of autogenous bone graft combined with plate fixation in total knee arthroplasty(TKA) with severe proximal medial tibial bone defect. METHODS: From March 2012 to October 2018, 21 patients (9 males and 12 females) with severe bone defects in the proximal medial tibia during primary total knee arthroplasty were treated with autogenous structural bone grafting and steel plate fixation, with an age of 61 to 77 years old with an average of (69.6±9.1) years and a course of 64 to 257 months with an average of (73.6±170.7) months. According to Rand classification, there were 13 cases of type Ⅲb and 8 cases of type Ⅳb. Postoperative complications were observed, and knee joint function was evaluated by the Hospital for Special Surgery (HSS) score and SF-36 quality of life score. RESULTS: All 21 patients were followed up for 37 to 64 months with an average of (49.5±13.7) months. The incisions of all patients healed smoothly, and 2 patients developed lower limb intermuscular venous plexus thrombosis after operation. There were no periprosthetic infection, loosening of prosthesis and other complications. The autogenous bone grafts of all patients achieved bony healing during postoperative X-ray follow-up, and the healing time was 8 to 13 months with an average of (10.1±2.3) months. The HSS score of patients increased significantly from 30 to 48 with an average of (53.4±4.2) before operation to 75 to 92 with an average of (81.2±8.4) at the final follow-up (P<0.05). The SF-36 quality of life score of patients after operation was significantly different from that before operation (P<0.05). CONCLUSION The technique of autogenous bone graft combined with steel plate fixation can achieve satisfactory osseointegration effect in the treatment of severe proximal tibial bone defects in primary knee arthroplasty, with less complications and obvious improvement in knee function.
Male ; Female ; Humans ; Middle Aged ; Aged ; Arthroplasty, Replacement, Knee/methods* ; Tibia/transplantation* ; Bone Transplantation/methods* ; Quality of Life ; Transplantation, Autologous ; Steel

Objective To investigate the value of serum lactate dehydrogenase (LDH) in advanced non-small cell lung cancer (NSCLC) treated with epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI).Methods Pretreatment LDH level,pathological characteristic,tumor staging and treatment situation of 190 advanced NSCLC patients with EGFR sensitive mutation confirmed by pathology were retrospectively collected in Zhuhai People's Hospital of Guangdong Provice from July 2011 to July 2015.All the patients were divided into LDH normal group (LDH ≤252 U/L,n =78) and elevated group (LDH ＞ 252 U/L,n =112) according to pretreatment LDH level.Inaging evaluations of the patients were performed regularly,and the progression-free survival (PFS) and overall survival (OS) were recorded.The survival curves were plotted by Kaplan-Meier method and survival difference between patients with different LDH level was compared by logrank test.Cox regression analysis was used to analyze prognostic factors for mortality.Results The objective response rate of the LDH normal group was 76.9％ (60/78),and the elevated group was 71.4％ (80/112),with no statistically significant difference (x2 =0.716,P =0.398).The disease control rate of the LDH normal group was 89.7％ (70/78),and the elevated group was 85.7％ (96/112),with no statistically significant difference (x2 =0.676,P =0.411).The median PFS of the LDH normal group was 11.5 months,and the elevated group was 9.7 months (x2 =5.92,P =0.015).The median OS was 31.0 months in the LDH normal group,and 26.1 months in the elevated LDH group (x2 =4.79,P =0.029).Both PFS and OS of patients with elevated LDH were shorter than those of patients with normal LDH.Cox multivariate regression analysis showed that tumor staging (HR =1.652,95％ CI:1.386-2.259,P =0.018),PS score (HR =2.248,95％ CI:1.507-3.846,P ＜ 0.001),carcino-embryonic antigen (CEA) level (HR =1.250,95％ CI:1.066-1.703,P =0.037) and LDH level (HR =1.771,95 ％ CI:1.324-1.947,P =0.015) were independent prognostic factors in patients with advanced NSCLC.Conclusion Pretreatment serum LDH can not affect the objective response rate and disease control rate of EGFR-TKI in the treatment of advanced NSCLC,but can affect the PFS and OS of patients.Pretreatment serum LDH is an independent prognostic factor.

Objective To develop a chemiluminescence imaging DNA microarray method for simultaneous,quick and accurate detection of serotypes of human adenovirus (HAdV ),namely,HAdV3,HAdV7,HAdV11,HAdV14 and HAdV55.Methods Based on the specific gene sequences in the conserved region of adenovirus from GenBank, oligonucleotide primers and probes were designed and synthesized to prepare the oligonucleotide microarray.The specific genomic sequences were amplified by multiplex PCR method.The multiplex PCR amplification products were hybridized with the specific probes of microarray that was scanned after washing and chemiluminescenceb before the result was analyzed.After optimization of the multiple PCR system,hybridization reactions and conditions of chemiluminescence,the specificity,sensitivity and reproducibility of the chip were evaluated.Results The microarray displayed high sensitivity, specificity and reproducibility.The minimum detection limit of plasmidg DNA was 3 ×103 copies/reaction.The microarray detection results of 38 clinical samples were approximately consistent with those using the direct sequencing method(37 /38).Conclusion A chemiluminescence imaging DNA microarray method for quick,sensitive and specific detection of five serotypes of HAdV is established,which can provide a new means for detecting serotypes of HAdV.

Objective To develop a detection method based on the technology of gene chips which can quickly distinguish genes of Enterococcus faecalis, E.faecium and vancomycin resistance.Methods Based on the specific gene ( ddl) sequences of two types of Enterococcus from GenBank, oligonucleotide probes which could detect and distinguish special genes and drug resistance genes ( vanA,vanB) of Enterococcus were designed and compounded.Then,the probes were dotted to modified slide.The target DNA fragments of vancomycin-resistant Enterococcus ( VRE) were labeled with biotin by multiple PCR amplification, and then hybridized with oligonucleotide probes on slide.The results were analyzed by portable imager.The multiple PCR system, hybridization reaction and condition of the chemiluminescence method were optimized before the specificity, sensitivity and reproducibility of the chip were evaluated.Results One universal primer, four specific primers, one universal probe and four specific probes were selected.This gene chip was demonstrated of high specificity and repeatability.The detection sensitivity was 103 CFU/ml.The gene chip detection results of 10 clinical samples were basically consistent with the drug sensitivity test ( 8/10 ) .Conclusion A gene chip technique for the detection of VRE is established successfully.It is possible to distinguish the type of VRE and detect the genetic phenotypes of drug resistance by gene chip technique.

Objective To evaluate the practical efficacy of comparison method in experimental teaching of pathological anatomy.Methods The junior college students of stomatology were divided into experimental group (n =48) and control group (n =48) randomly.Students in experimental group adopted the experimental teaching with comparison method of pathological anatomy and histology while those in control group followed the traditional experimental teaching.Experiment examination results of two groups were compared and questionnaire investigation was carried out among students in experimental group.Results Average score of experiment examination in experimental group was (88.35 ±5.68) while that in control group was (78.35 ±4.86),with statistical differences (P ＜0.01).The results of questionnaire investigation showed that students in experiment group preferred comparison teaching method.Conclusions Comparison teaching method is feasible,the mechanism of which emphasizes gaining new knowledge by reviewing the old and strengthens the contact of basic subjects,therefore it is conducive to enhancing the quality of pathological experimental teaching.

Objective To analyze the association between β2-AK 27 locus genetic polymorphisms and asthma, and the protective effect in airway hyperreactivity. Methods The allel polymerase chain reaction were used to determine β2-AR 27 locus genetic polymorphisms in 149 patients with cough variant asthma who have the airway hyperreactivity. To observe these people for two years in order to know the proportion of changed to typical asthma. And compare with 90 people in healthy group. Results (1) The distribution frequency of β2-AR 27 locus genetic polymorphisms is major in heterozygote (57 % ) , and the Glu/Glu homozygote has the least ( 20% ) , (2) There was a significant decrease in the frequency of Glu/Glu genotype in asthmatics compared with healthy group(9% VS 20% ) ,OR = 0.4(P<0.05) ,95% CI (0.2 ～0. 9) ,but there was no significant difference in the allele frequency of asthmatics compared with healthy group,(3)The frequency of Glu/Glu genotype in severe asthma was lower than stable asthma group(P<0. 05). Conclusion These results suggesteded that β2-AR 27 locus genetic polymorphisms is correlated with asthma,and the Glu27 could have the protective effect to the airway.

Objective To measure and evaluate mesenteric artery by comparing the multi-slice spiral CT mesenteric artery images with autopsy specimens. Methods Totally 230 normal subjects were selected to undergo abdominal multi-slice spiral CT and enhanced CT. We processed the images;R3econstructed 3D images, analyzed and compare the mesenteric artery images obtained by multi-slice spiral CT with autopsy specimens. Results 1. Diameters of mesenteric artery obtained by vivo image were significantly larger than that of autopsy specimens (P<0.05);2. Start locations, branch types and running directions of both superior mesenteric artery and inferior mesenteric artery were much different between traditional autopsy specimens information and our results.3. Different reconstruction methods had different advantages. Especially, STS-MIP method could present the level of mesenteric artery better. Conclusion The method for mesenteric artery study using multi-slice spiral CT can enhance scanning and 3D reconstruction with workstation has been approved to work well, and it is superior to traditional autopsy specimen method. It is also convenient for mesenteric artery scientific evaluation. The result data of this method are reliable. Moreover, this method is available to research with large number of specimen.

Objective To investigate the clinical pathological feature and treatment of cystosarcoma phyllodesand.Methods 12 cases of cystosarcoma phyllodesand after surgical were retrospectively reviewed.Results Among the 12 cases cystosarcoma phyllodesand,5 cases underwent total mastectomy,2 cases underwent modified radical mastectomy,1 case underwent local excision and reserve nipple,and 4 cases underwent lesion removed and reserve nipple or gland.All cases were follow up between 1～18 years,1 case lost,2 cases recurred after surgical 1.5 and 2.0 years and cystosarcoma were removed again,and other patients were free from cancer cell.Conclusion The diagnosis and treatment of the cystosarcoma phyllodesand are based on the pathology.It is difficult to predict the behavior by the pure histology from the cell appearance,recurrence of the cystosarcoma phyllodesand is relevant with the different classification tumor and size.

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj14. The expression of Sj14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.