Heavy chain antibodies (HcAb) without light chains are naturally produced by camelids.The single domain antigen-binding fragment of HcAb is referred to as VHH or nanobody (Nb),which is the smallest antigen-binding entity at present.Several characteristics such as low molecular weight,high stability,and low immunogenicity make the use of nanobodies superior to the conventional antibodies.Currently,nanobodies are highly valuable antibodies for various applications,including fundamental research,diagnostics,and therapeutics.A variety of nanobodies are investigated in clinical researches.The recent progresses of heterologous expression of nanobodies in seven expression systems,including Gram-negative and positive bacteria,yeasts,filamentous fungi,insect cells,mammalian cells and plant cells were focused on,and the expression systems,hosts,characteristics of vectors,construction of vectors and yield of nanobodies were summarized.The strategies to increase the yield of nanobodies are discussed from molecular level,expression level and rational design.

Objective To evaluate the image quality of dynamic three-dimensional contrast enhanced ultrasonography(3D-CEUS) in liver tumors.Methods Twenty-five patients were examed with 2D-CEUS and dynamic 3D-CEUS.Contrast-enhanced computed tomography or magnetic resonance imaging(CECT/MRI) were performed early or late.Two readers reviewed the images of both modalities and graded the images according to diagnostic criteria as follows:tumor blood fusion in arterial phase(unclear-score 0,moderate-score 1,clear-score 2),tumor boundry and relationship with periphery tissues (unclear score 0,moderate-score 1,clear-score 2).The concordance between the two modalities for showing liver tumors was evaluated by Kappa test.Results Twenty-three of 25 danymic 3D-CEUS were displayed successfully versus total objects in CECT/ MRI.Among them,for showing blood fusion,16 lesions scored 2,7 lesions scored 1,2 lesions scored 0 in danymic 3D-CEUS,meanwhile 18 lesions scored 2,7 lesions scored 1,0 lesion scored 0 in CECT/MRI,the Kappa value was 0.65.For showing tumor boundry and relationship with peripherytissues,16 lesions scored 2,7 lesions scored 1,2 lesions scored 0 in danymic 3D-CEUS,meanwhile 21 lesions scored 2,4 lesions scored 1,0 lesion scored 0 in CECT/MRI,the Kappa value was 0.52.Conclusions Danymic 3D-CEUS was shown to be useful for displaying tumor blood fusion,boundry and relationship with periphery tissues in single liver tuomor,the concordance between dynamic CEUS and CECT/MRI for single liver tuomor was fair.For huge liver tumor and multi-tumors,3D-CEUS can not give overall information.

Objective To explore the preventive effect and mechanism of pioglitazone on nonalcoholic fatty liver disease in rats.Methods Thirty six male Sprague-Dawley ( SD) rats were randomly divided into normal control group , high-fat diet group , and pioglitazone intervention group , with 12 rats in each group .Normal control group fed with standard diet for eight weeks , and the rest two groups fed with high fat diet .Pioglitazone intervention group fed with pioglitazone in the four to eight weeks , the rest two groups in the same period fed with distilled water .Fasting serum alanine aminotransferase , aspartate aminotransferase , total cholesterol , triglyc-erides, fasting glucose , and fasting blood insulin levels were determined .The fasting insulin resistance index was calculated .Routine histological features of hepatic section were observed by hematoxylin eosin (HE) staining.The shape change of Kupffer cells in the liv-er was detected .The levels of tumor necrosis factor ( TNF)-αand nitric oxide ( NO) secreted by Kupffer cells were measured .Results The fasting insulin resistance index , triglycerides , and total cholesterol in rats of high-fat diet group were significantly higher than in normal control group ( P <0.05 ) .Histopathological examination showed hepatocellular macrove sicular steatosis , lobular inflammatory cell infiltration, and inflammatory cell infiltration .Hepatic Kupffer cell morphology change , its produced TNF, NO and liver tissue pa-thology changes were positively correlated ( P <0.05 ) .The fasting insulin resistance index , triglycerides , total cholesterol was signifi-cantly lower in rats of pioglitazone intervention group than the high-fat diet group ( P <0.05 ) .The liver tissue structure was obviously improved pioglitazone intervention group rats , morphology and function of liver Kupffer cells were normal .Conclusions Pioglitazone can prevent the nonalcoholic fatty liver disease induced by high fat diet .The mechanism may be related to improve insulin resistance , reduce blood fat , and regulate function of Kupffer cells .

Actinic keratosis (AK) is a precursor to cutaneous squamous cell carcinoma (SCC),and molecular mechanisms of transformation from AK to SCC have been a research hotspot.This review focuses on four aspects of the molecular mechanisms,including chromosomal variation,gene mutations,signaling pathways and other factors,summarizes advances in selective mutations in chromosomes 3p,9p,9q,13q,17p and 17q,heterozygous deletion mutations in chromosome 18q,mutations of p53,p16 and Ras genes,abnormality of the transforming growth factor β1 signaling pathway and Fas/FasL signaling pathway,and aberrant expression of matrix metalloproteinases in the transformation from AK to SCC,which will provide a reference for further researches.

In order to promote rehabilitation exercises for breast cancer patients and to improve their quality of life,as well as to promote the development of nursing profession, our hospital has estab-lished a specialistnursing clinic for breast cancer since January 2016. We standardized nurses' qualification and training requirements, and performed management in patients'symptom assessment, nursingtechniqueguidance for breast cancer, sleep guidance, health education, long-term follow up, etc. After one year of implementation, symptom assessment was conducted 1283 times, rehabilitationexerciseguidance was 329 times, lymphatic drainage guidance was 297 times, sleep guidance was 159 times, health education was 591 times, sexual life with spouse guidance was 107 times, in-hospital nursing consultation was 48 times, family on-site demonstration was 29 times. The satis-faction rate was 98.4% which was higher than average level of the hospital according to outpatient investigation.

Objective To study the relatinonship between the regional cerebral blood flow (rCBF) and NF-kB expression in perihematoma region after intracerebral hemorrhage (ICH).Methods 27 healthy dogs were divided into control group (3 cases) and ICH group (24 cases) at random. The models of ICH were made by injecting self-blood into the frontal lobes. At different points (3 h, 6 h, 12 h, 24 h, 48 h, 72 h, 7 d and 15 d), the changes of rCBF in the region of perihematoma were observed by perfusion weighted imaging (PWI). Immunohistochemistry SP method was used to detect expressions of NF-kB. Immune positive cells were stained brown-yellow colour in the cytoplasm and/or nucleus, and NF-kB positive particles that transformed from cytoplasm to nucleus showed activation of NF-kB. Results In the region of perihematoma, PWI showed hypo-perfusion within 12h, reperfusion or hyper-perfusion from 12 to 24 h, and slightly hypo-perfution after 48h. NF-kB positive cells presented at 6 h, reached the peak from 12 to 48 h, decreased from 72 h to 7 d, nearly disappeared at 15 d. NF-kB positive cells mainly existed in the region of perihematoma. From 12 to 48 h, NF-kB expression was markedly in quantities of neurons and gliacytes. The activated NF-kB was clearly shown at the same time.Conclusions Cerebral ischemia and reperfusion in the region of perihematoma could induce expression of NF-kB, which was involved in the inflammatory and immune reaction and resulted in the ingury caused by cerebral ischemia and reperfusion.

AIM: To investigate the correlation of hepatitis B virus X protein (HBx) with renal tubular epithelialcell apoptosis in hepatitis B virus-associated glomerulonephritis (HBVGN) and the possible signaling mechanism. METHODS: The activation of JAK2/STAT3 signal pathway and the expression of apoptosis -related proteins in humankindey proximal tubular epithelial cells (HK-2 cells) were determined by Western blotting after transfection with HBx eukaryoticexpression vector.The cell proliferation was observed by CCK-8 assay.The cell apoptosis was analyzed by the imagingof HO33342 staining, transmission electron microscopy and flow cytometry with Annexin V /PI double staining.RESULTS:After transfection of the target gene HBx, the expression levels of both p-JAK2 and p-STAT3 were significantly increased.At the same time, the cell proliferation was obviously inhibited, and the apoptotic rate was increased.After incubationwith AG490, the JAK2/STAT3 signal pathway was partially blocked, and the cell apoptosis induced by HBx was reduced. CONCLUSION: HBx up-regulates the activation of JAK2/STAT3 signal pathway to induce renal tubular epithelialcell apoptosis, which is possibly involved in the pathogenic mechanism that HBV directly damages nephridial tissue .

BACKGROUND:Immune rejections after organ transplantation or serious adverse reactions due to immunosuppressive drugs show a lack of effective treatments and poor therapeutic outcomes. Therefore, we try to find an effective immune suprresion method in combination of the latest immunomodulatory achievements. OBJECTIVE:To construct a lentiviral vector overexpressing indoleamine 2,3-dioxygenase (IDO). METHODS:(1) The IDO gene that was successful y contructed was inserted into lentiviral packaging plasmids GV308 to construct GV308-IDO lentivirus packaging plasmids. (2) The 293T cel s with 80%confluence were co-cultured with 5'LTR and 3'LTR, basic elements of lentiviral packaging auxiliary components, including Psi, cPPT, 3FLAG, TetR, IRES, WRPE, TetIIP, Ubiquitin Promoter, SV40 origin and HIV. RESULTS AND CONCLUSION:Western blot assay showed that in 10 g/L agarose gel electrophoresis, there was a target fragment at Mr 48 000. This value was consistent with the size of IDO protein. RT-PCR results showed visible IDO expression in 293T cel s. These findings suggest that IDO fusion gene has been successful y reorganized in the lentiviral packaging plasmids.

Objective To explore the clinical characteristics and prognosis factors of child hemophagocytic syndrome(HPS)in different age groups.Methods Eighty-five children cases of HPS were divided into 3 age groups,0-2 years old (group A),＞2-8 years old (group B) and ＞8-14 years old (group C).The etiology,pathology,clinical symptoms,laboratory detection and prognosis were retrospectively analyzed.Results In the etiology,the etiology in the group A,B and C was dominated by EB virus infection,the infection positive rate in the group C was significantly higher than that in the group B and C;in the bone marrow examination:the occurrence rate of granulocytes to erythrocytes ratio ＜1.0 in the group A was higher than that in the group C;in the clinical symptoms,cervical lymph node enlargement mostly occurred in the group B,hepatosplenomegaly was most rare in the group C,and the serous cavity effusion commonly occurred in the group B;in the laboratory detection,the LDH level in the group B was higher than that in the group A,the above differences were statistically significant(P＜0.05).There were no statistically significant differences in other clinical symptoms and laboratory detection results among the three groups (P＞0.05);In the comparison between the survival group and death group,the serous cavity effusion,triglyceride(TG),CD4/CD8 ratio and granulocytes to erythrocytes ratio in bone marrow had statistical difference(P＜0.05).The above factors were performed the Logistic analysis,the results showed that TG＞3.5 mmol/L,CD4/CD8 ratio ＜1.24 and bone marrow granulocytes to erythrocytes ratio ＜1.02 were the unfavorable risk factors affecting the prognosis of child HPS (P＜0.05).Conclusion Partial clinical characteristics of child HPS are related with the onset age,TG level,CD4/CD8 ratio and bone marrow granulocytes to erythrocytes ratio are the risk factors affecting prognosis.