Tissue blotting, based on Enzyme-Linked Immunosorbent Assay (ELISA), is a technique for detection of plant viruses. This technique is not only high sensitivity and specificity, but also simpler and more rapid for detection. Samples that are blotted on membrane can be kept over three months. The results can directly display the section of virus infected. It is especially suitable for detection of plant viruses on a large scale.

Objective To evaluate the clinical efficacy of transjugular intrahepatic portosystemic shunt(TIPS)combined with stomach and esophageal variceal embolization(SEVE)for gastric variceal haem-orrhage,and the efficacy with or without a gastrorenal shunt. Methods A total of 52 patients with gastric variceal bleeding history and portal hypertension treated with TIPS combined with SEVE were included from October 2013 to March 2015.Patients were divided into two groups according to preoperateive CT angiogra-phy,27 cases with gastric variceal haemorrhage associated with a gastrorenal shunt in group A,and 25 gastric varices bleeding cases without gastrorenal shunt in group B. During the follow-up,the incidence of the total rates of rebleeding,TIPS primary patency and hepatic encephalopathy,and the survival rates were compared between group A and group B. Results In all patients,the average portal vein pressure decreased from 36. 50±7. 00 cmH2 O(1 cmH2 O= 0. 098 kPa)before operation to 28. 15±6. 27 cmH2 O after TIPS combined with SEVE,with significant difference(t= 10. 357,P= 0. 001). Fifty two patients were followed up for 1 to 18 months(1-18 months in group A;1-15 months in group B).The total rates of rebleeding,TIPS primary patency,hepatic encephalopathy and survival were 11. 54%(6/ 52),86. 54%(45/ 52),11. 54%(6/ 52) and 92. 31%(48/ 52),respectively. There were no significant differences between the two groups in the total rates of rebleeding[11. 11%(3/ 27)VS 12. 00%(3/ 25),P = 1. 000],TIPS primary patency[88. 89%(24/ 27)VS 84. 00%(21/ 25),P= 1. 000],hepatic encephalopathy[14. 81%(4/ 27)VS 8. 00%(2/ 25), P= 0. 738]or total survival rate[92. 59%(25/ 27)VS 92. 00%(23/ 25),P = 1. 000]after TIPS combined with SEVE. Conclusion TIPS combined with SEVE is effective for gastric varices,and equally effective in the treatment of both gastric variceal haemorrhage associated with a gastrorenal shunt and gastric varices bleeding without gastrorenal shunt.

BACKGROUND: For xenogeneic bone transplantation, immune rejection is the major problem that affects the prognosis. However, the understanding about the expression and regulation of the immune factors in heterogenic bone transplantation is limited. Interleukin-1 (IL-1), interleukin-6 (IL-6)and tumor necrosis factor alpha (TNF-α) are important immune factors, and are closely related with post-transplantation rejection.OBJECTIVE: To observe the regional expressions of mRNA and protein of IL-1α, IL-6 and TNF-α in xenogeneic bone transplantation, and to in vestigate the effect of transforming growth factor (TGF)-β on these immune factors.DESIGN: A randomized controlled experiment. SETTING: The Orthopaedic Institute of Xijing Hospital of the Fourth Military Medical University of Chinese PLA MATERIALS: Totally 72 male Balb/c mice, with a body mass of 20 to 25 g, were randomly divided into 3 groups: combined bovine cancellous bone (bCB) granule group (Group A), simple bCB granule group (Group B) and blank control group (Group C) with 24 mice in each group.INTERVENTIONS: This experiment was conducted at the Institute of Orthopaedics, Xijing Hospital, Fourth Military Medical Univessity of Chinese PLA from June 2003 to June 2004. In Group A, one bCB combined TGF-β was implanted into the muscles of left thigh of each mouse. In Group B, one bCB alone was implanted, and in Group C, no bCB was implanted. The number of proliferation of cells in bone implantation area or adjacent tissues of the operated area was observed at postoperative 4, 7, 14 and 21 days; and the regional expressions of several immune factors in im plants were detected with in situ hybridization and immunocytochemistry methods.MAIN OUTCOME MEASURES: Histological observation and detection of regional cellular density of the implants of the mice in each group; the regional expressions of mRNA and protein of IL-1α,IL-6 and TNF-α in the implants RESULTS: ①) Histological observation and detection of regional cellular density of the implants of the mice in each group: on day 7, the cellular density of the proliferated tissues was significantly higher in the Group A than in the Group B [(470.63±132.89), (311.46±93.69)/field,P ＜ 0.01];But on days 14 and 21, there was no significant difference. ②The regional expressions of mRNA and protein of IL-lα, IL-6 and TNF-α of the im plants of the mice in each group. On days 7 and 14 after xenografts were implanted, the regional expressions of IL-1α, IL-6 was respectively lower the xenografts combined with TGF-βthan in the simple xenogeneic bone (day 7: IL-1α 42.55±9.65 vs 67.95±17.82,IL-6 48.26±11.17 vs 77.21±15,16;day 14: IL-1α mRNA 84.77 ±7.42 vs 112.94±7.02,78.1 ±17.22 vs 121.18±15.44,P ＜ 0.01) ,but for the TNF-α, there was no significant difference (P ＞ 0.05).CONCLUSION: In the region of bone xenograft, a variety of cells express mRNAs and proteins of IL-1α, IL-6 and TNF-α, and their expressions are regulated by TGF-β. It may imply a kind of regulation of the immunity of bone xenograft by TGF-β.

Objective To characterize the clinical characteristics of lupus mesenteric vasculitis (LMV). Methods Analyzing the clinical, laboratory and treatment data of LMV patients hospitalized from 2002. 1.1 to 2007. 12. 31 retrospectively. Results (1) The three common manifestations were abdominal pain, diarrhea and vomit with the prevalence rate of 77%, 70% and 67% respectively. (2)The majority of LMV cases were active vital organ (28/30), kidney (24/30) and hematological system (18/30) were the main organs of involvement. Ten patients had hydroureteronephrosis, and 8 patients had intestinal pseudo-obstruction at the same time. (3) Systemic lupus erythematosus disease activity index (SLEDAI) score was ≥10 in 80% (24/30) of patients. The progression of LMV was accompanied with new-onset ieucopenia or worsening leucopenia or hypocomplementemia in 10 cases. (4) Blood antinuclear antibodies were positive in 27 patients detected, and anti-SSA antibody was positive in 15 (56%), anti-U1RNP antibody was positive in 14 (52%). (5) Fourteen cases had bowel wall thickening with target sign or mesenteric vessels with palisade or comb sign in contrast CT scan of abdomen. (6)Twenty-seven cases were treated with orally or intravenous medium to high dose steroid therapy and recovered from LMV. Conclusions (1) Abdominal pain, diarrhea and vomit were frequent manifestations of LMV patients. (2) LMV was one of the serious complications of systemic lupus erythematosus(SLE), and usually accompanied by active SLE in other organs. (3) A drop in the white blood cell count or complement C3 titer might be correlate with the occurrence of LMV. It needs to further investigate the relationship between LMV and the high positive rate of anti-SSA and anti-U1RNP antibody. (4) LMV patients responded well to intravenous high dose methylprednisolone.

OBJECTIVE: To explore the effect of the traditional Chinese medicine Yifei Kangliu (YFKL) Oral Liquid on the proliferation, cell cycle and protein-nucleic acid synthesis of murine Lewis lung cancer cell and human lung adenocarcinoma cell line SPC-A-1. METHODS: The inhibiting rates of tumor growth were calculated by weighing the weight of tumor inoculated in vivo, combined by counting cancer cells in vitro. The ratio of the cell cycle and exponents of DNA, RNA, and protein were measured by flow cytometry (FCM). RESULTS: The inhibiting rate of tumor growth in the treated group with YFKL Oral Liquid was 30.38% (P<0.05). The proportion of cells in S phase of the treated groups with YFKL Oral Liquid was lower than that of the control group. In the group with most significant result, 72% of the cells were stagnated in G0/G1 phase. The inhibiting rates of DNA, RNA and protein in murine Lewis lung cancer were 7.4%, 23.73% and 23.31% respectively. In SPC-A-1 cell line, the inhibiting rates were 9.3%, 10.1% and 14.7% respectively, demonstrating amplified effects on lower levels. CONCLUSION: YFKL Oral Liquid significantly inhibited the proliferation of murine Lewis lung cancer cell and human lung adenocarcinoma cell line SPC-A-1 by blocking the cancer cells entering the proliferative phase resulted from its inhibition of DNA.

Objective To investigate the immunological properties of Rv1009 domain. Methods BALB/c mice were immunized with Rv1009 domain three times at 2-week interval. ELISA was used to detect the antiRv1009 domain antibody titer in the sera of immunized mice sera. The spleen lymphocytes of the immunized mice were separated and the stimulation index (SI) was measured by MTT colorimetry. Levels of secreted IFN-γ, IL-10 and IL-12 upon specific antigen stimulation were detected by ELISA. The BALB/c mice immunized with Rv1009 domain were intravenously infected with MTB H37Rv. Four weeks after the final injection, the number of CFU in spleens was determined. Results The titer of the specific antibody in sera of the immunized BALB/c mice was 1:12 800. The SI of Rv1009 domain immunized group (2. 40±0. 18) was significantly higher than that of saline immunized group (0.90±0.21). The IFN-γ,IL-10 and IL-12 levels in culture supematant of spleen lymphecytes from the fusion proteins immunized mice was (1 432±30) ng/L, (503±11) ng/L and (311±11) ng/L respectively, significant different from that of saline immunized group[(256±20) ng/L, (76±6) ng/L and(56±8) ng/L,P<0.01]. Four weeks after the final injection,compared with normal saline immunized mice (6.64±0.13), dramatic reduction in MTB replication was observed in the spleen (4.86±0.14) from BALB/c mice immunized with fusion proteins following a subsequent MTB H37Rv challenge, but the protection efficacy of mice immunized with Rv1009 domain was not as good as that of BCG vaccination group (3.81±0.16). Conclusion Rv1009 domain can be used as a candidate for the new TB vaccine.

Objective To evaluate the efficacy and safety of transjugular intrahepatic portosystemic shunt (TIPS) combined with gastroesophageal variceal embolization (SEVE) in the treatment of patients with gastroesophageal varices accompanied by a gastrorenal shunt with a diameter over 5 mm .Methods From October 2013 to December 2014 ,the clinical data of 18 patients with portal hypertension caused gastroesophageal variceal bleeding and treated by TIPS combined with SEVE were collected . The difference of portosystemic pressure gradient between before and after operation was observed ,and Child‐Pugh score before and after operation was also evaluated .The hemostasis in 24 hours ,TIPS patency ,the occurrence of hepatic encephalopathy(HE) ,rebleeding ,hepatic failure ,mortality and the change of liver function and thrombocytopenia were recorded after operation .Student′s t‐test or analysis of variance was performed for statistical analysis of measurement data .Results In the 18 patients ,the average portal vein pressure decreased from (34 .23 ± 6 .35) cmH2O (1 cmH2O= 0 .098 kPa) before operation to (25 .69 ± 6 .89) cmH2O after TIPS combined with SEVE ,and the difference was statistically significant (t=7 .572 , P<0 .01) .The difference of portal vein pressure before and after operation was (8 .54 ± 4 .79) cmH2O . No hepatic failure was observed in all 18 patients .Among 10 patients with emergency operation ,nine patients obtained successful hemostasis in 24 hours .No operation related complications were observed in all the patients .During the follow‐up period of 18 patients ,two patients had variceal rebleeding confirmed by endoscopy ;one patient had stent stenosis;three patients got HE and one patient died .At different follow‐up time point of patients ,there was no significant difference in Child‐Pugh score ,blood ammonia level ,albumin level ,bilirubin level ,white blood cell count and blood platelets count compared with those before operation (all P>0 .05) .Conclusion TIPS combined with SEVE in the treatment of patients with gastroesophageal varices accompanied by a gastrorenal shunt with a diameter over 5 mm could effectively control bleeding ,and no ectopic embolism happened .

Objective To explore the brain function changes in cirrhotic patients with normal blood ammonia and high in direct bilirubin with regional homogeneity (ReHo) of resting-state functional MRI (rs-fMRI).Methods Fifty-three cirrhotic patients with normal blood ammonia and high indirect bilirubin (cirrhosis group) and 60 healthy volunteers (control group) performed rs-fMRI scans.ReHo values in different brain areas of the two groups were acquired and compared.The correlation between the ReHo values of different brain regions in both groups and the indirect bilirubin concentrations was analyzed.Results Compared with control group,cirrhosis group showed that ReHo values increased in left dorsal-lateral frontal gyrus,left orbital superior frontal gyrus,left top edge of angular gyrus,left precentral gyrus,right caudate,right putamen,right insula,right olfactory cortex (all P＜0.001),and decreased in left superior occipital gyrus,left middle occipital gyrus,left cuneus,left orbital inferior frontal gyrus,right paracentral lobule,right precuneus,right middle occipital gyrus,bilateral lingual gyrus and bilateral cerebellum (all P＜0.001).ReHo values in right insula and right putamen in cirrhotic patients positively correlated with indirect bilirubin (r=0.32,P=0.021).Conclusion The resting state brain function increases and decreases in some brain regions of cirrhotic patients with normal blood ammonia and high indirect bilirubin.Indirect bilirubin level correlates with function of some brain regions,which has certain effect on brain function of adult.

Objective To detect the isoniazid (INH) and rifampin (RIF) resistance of Mycohaeterium tuberculosis isolates in the single tube with multiplex-polymerase chain reaction-single strand conformation polymorphism(muhi-PCR-SSCP) system. Methods According to the sequences of inhA, katG and rpoB genes of the Mycohacterium tuberculosis, three pairs of oligonucleotide primes were designed to examine the INH and RIF resistance with the multi-PCR-SSCP. The validity of the newly developed method was evaluated with 116 clinical isolates of Mycohacterium tuberculosis( 70 isolates that were INH-resistant and 66 isolates that were RIF-resistant). Results The validity of the method was assessed with multiplex PCR-SSCP with the bacteria culture with susceptibility test as golden standard. The three genes, katG, inhA and rpoB, in the 116 clinical isolates and H37Rv strain were amplified successfully in single PCR reactions,except 4 isolates with katG deletion mutants. Compared with strain H37Rv, forty-six isolates had katG gene mutations, thirteen had inhA mutations and fifty-eight had rpoB mutations. Thirty-eight isolates had simultaneous katG and rpoB mutations and 4 isolates had both inhA and rpoB mutations. Four isolates had inhA and katG mutations and 2 isolates had mutations in all three genes simultaneously. The sensitivity of the newly developed multiplex-PCR-SSCP assay was 80% and 82% for INH and RIF, respectively. The specificity of the assay was 100% and 92% for INH and RIF, respectively. Conclusion Muhiplex-PCRSSCP provides a rapid, specific and cost-effective method of detecting multidrug-resistant TB. It laid a solid foundation for the further study of drug resistant gene.

Objective To investigate the correlation between admission hyperglycemia and clinical prognosis in elderly patients with community-acquired pneumonia.Methods Based on the electrical medical record (EMR),information of the patients with pneumonia was retrospectively collected and the necessary data were prepared and arranged.Based on clinical outcomes,demographic characteristics and clinical indicators were compared between groups in order to screen potential variables affecting clinical outcomes.Then,the random blood glucose level 8.6 mmol/L was considered to be the cutoff point for grouping patients,with clinical outcomes as outcome variables,random blood glucose levels as grouping variables,propensity score value analysis method was applied along with the ratio of 1:1 nearest neighbor matching method to generate paired samples in order to assess the impact of blood glucose level on mortality rate;With hospitalization time serving as time variable,survive analysis method was used to analyze COX proportional model.Results Five hundred and fifty-eight patients were enrolled in the analysis,197 patients of which were recorded dead during hospitalization (the dead group).After the correction of demographic characteristics,past medical history and blood test indexes,white blood cells,C reactive protein,random blood glucose at admission and triglyceride levels were regarded as potential factors for the deaths of patients (P<0.05).However,the propensity score analysis method suggested that there was no significant difference between the blood glucose levels and mortality.The survival analysis showed that the mortality in the high blood glucose group was higher than that in the normal blood glucose group (HR=1.68,95%CI(1.22,2.31)).Conclusion Admission hyperglycemia is an important indicator of adverse prognosis in elderly patients with pneumonia.It may be of significance for screening and identifying high-risk patients.