OBJECTIVETo explore the effect of Bushen Huoxue Compound (BHC) on lactate dehydrogenase (LDH) leakage, expressions of vascular endothelial growth factor (VEGF) and VEGF mRNA in retinal Muller cells under high glucose condition or advanced glycosylation end products (AGEs) condition by using serum pharmacological method.

METHODSThe retinal Müller cells of 5-7 days post-natal Sprague Dawley (SD) rats were cultured with modified enzyme-digestion method. Purified retinal Muller cells were cultured in normal conditions, high glucose condition (50 mmol/L) or AGEs (50 mg/L and 100 mg/L) conditions, and BHC-containing serum was added to culture medium. The LDH leakage and VEGF expressions were measured by enzyme-linked immunosorbent assay (ELISA). In addition, the relative expression of VEGF mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSCompared with the normal control group, expressions of VEGF and VEGF mRNA were significantly increased in the high glucose group, the low dose AGEs group and the high dose AGEs group (all P < 0.01). The LDH leakage was obviously increased in the high dose AGEs group, when compared with the normal control group and the high glucose group (P < 0.01). The LDH leakage, expressions of VEGF and VEGF mRNA were obviously decreased by BHC-containing serum both in high glucose and AGEs conditions (P < 0.05, P < 0.01). BHC-containing serum had no significant effect on the LDH leakage and expressions of VEGF and VEGF mRNA in normal conditions (P > 0.05).

CONCLUSIONSAGEs intervention could obviously lower the stability of Müller cell membrane. Up-regulated expressions of VEGF and VEGF mRNA in cultured Müller cells could be induced by AGEs or high glucose. BHC-containing serum could stabilize the stability of Müller cell membrane, inhibit the transcription of VEGF mRNA and decrease the protein expression of VEGF, which might be one of important mechanisms for preventing and treating diabetic retinopathy.

Animals ; Cells, Cultured ; Diabetic Retinopathy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Ependymoglial Cells ; Glucose ; L-Lactate Dehydrogenase ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A

Objective:To construct double copy and x gene deleted HBV expression plasmid and study its expression in Hep3B cell line. Methods:The double copy HBV DNA ( adr Ⅰ) was used to inactivate HBV x gene by inserting mutation and gene recombination. The inserted 55 bp DNA sequence was synthesized artificially; the insertion point was ApaL Ⅰ of x gene area. After recombination, an x gene defected HBV plasmid containing single P, S and C gene was constructed,which can express in mammalian cell line. Another plasmid carrying double copy HBV DNA with normal x gene was constructed as contrast. Both were used to transfect Hep3B cells. Then the cells were screened by G418 and HBV virus in culture medium were isolated and detected by fluorescence quantitative PCR. Results: Plasmids pcDNA3 KN F1F2 and pcDNA3 ES HBV2 were constructed successfully. After cell transfection, the HBV DNA was highly expressed with both plasmids on the 3 rd, 6 th,14th day. Conclusion: The plasmids constructed can express in Hep3B cell line and cause HBV replication; x gene defected HBV gene has no effect on HBV replication in Hep3B cell line.

Objective To investigate the effect of L-arginine on expression of protein kinase C(PKC)mRNA during pulmonary ischemia and reperfusion injury(PIRI)in the rabbits.Methods Single lung ischemia and reperfusion animal model was used in vivo.The rabbits were randomly divided into three groups(n=9,in each),sham operated group (Sham),PIR group(I-R)and PIR+L-arginine group(L-Arg).Changes of several rariables including malondialdehyde (MDA),superoxide dismutase(SOD),malandialde hyde(MDA),nitril oxide(NO),wet to dry ratio of lung tissue weight(W/D)and index of quantitative assessment(IQA)of histolngic lung injury were recorded at 60 minutes after reperfusion in lung tissue.Meanwhile the location and expression of PKC mRNA were observed.Lung tissue was prepared for light microscopic and electron microscopic observation at 60 minutes after reperfusion.Results In comparison with group I-R,PKC mRNA strongly expressed in intima and extima of small pulmonary artery as well as thin-waU vessels (mostly small pulmonary veins).The average optical density values of PKC-?,?and?mRNA in small pulmonary veins in L-Arg group had significance(all P＜0.01);SOD increased while MDA,W/D and IQA decreased at 60 minutes after reperfusion in lung tissue(P＜0.01 and P＜0.05).A morphologically abnormal changes of the lung tissue,were lessen markedly in L-Arg group.Conclusion L-arginine possess notably protective effects on PIRI in rabbits by activating PKC-?,?and?mRNA expression in lung tissue,raising NO level,dropping oxygen free radical level and decreasing lipid peroxidation.

OBJECTIVETo investigate the viral pathogens of acute respiratory infection (ARI) in hospitalized children from Suzhou of China.

METHODSThe nasopharyngeal aspirate samples were obtained from 1,668 hospitalized children with ARI. Common respiratory viruses, including respiratory syncytial virus (RSV), influenza virus A and B, parainfluenza viruses 1, 2 and 3 and adenovirus, were detected using direct immunofluorescence. Human metapneumovirus (hMPV) gene fragments were detected by RT-PCR.

RESULTSViral agents were identified in 597 cases (35.8%). RSV was the most frequent (17.6%). RSV infection is more common in children less than 1 year old. A highest detection rate of RSV was found during winter and spring. hMPV was detected in 10.6% of the cases, with a peak detection rate between March and May. Single viral infection was found in 561 cases (33.6%) and mixed viral infection in 36 cases (including 27 cases at age of less than 1 year). RSV and hMPV co-infection was common (n=22).

CONCLUSIONSRSV is common pathogen of ARI in children from Suzhou. RSV viral activity peaks during winter and spring. The children at age of less than 1 year are susceptible to RSV. hMPV is also an important pathogen of ARI, with a peak detection rate between March and May. Mixed viral infection is common in children less than 1 year old.

Acute Disease ; Age Factors ; Child ; Child, Hospitalized ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Metapneumovirus ; isolation & purification ; Nasopharynx ; virology ; Respiratory Syncytial Virus, Human ; isolation & purification ; Respiratory Tract Infections ; epidemiology ; virology ; Seasons

OBJECTIVETo report the therapeutic effect of reverse fasciocutaneous flap pedicled with perforator branch of anterior medial malleolus artery for soft tissue defect on the dorsal side of foot.

METHODSThe perforator branch was located under the guidance of Doppler flowmeter. The flap was designed along the saphenous neurovascular axis. Then the flap was transferred reversely with the perforator branch as rotation point.

RESULTSFrom Feb. 2002 to Mar. 2008, 12 cases were treated and followed up for 6 - 18 months. All the flaps survived completely. The flap size ranged from 13.5 cm x 3.0 cm to 8 cm x 3 cm. The perforator branch located at 0.5 - 1.5 cm anterior-inferior to the medial malleolus. Both the cosmetic and functional results were satisfactory.

CONCLUSIONSThe flap has a reliable blood supply and a flexible design. It is easily performed for soft tissue defect on the dorsal side of foot. It is a new type flap which combined neurocutaneous vascular flap with the perforator flap.

Adult ; Fascia ; transplantation ; Female ; Foot Injuries ; surgery ; Humans ; Male ; Skin Transplantation ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; innervation

OBJECTIVETo study the fractional anisotropy (FA) and the architecture of the optic radiation fiber tracts of normal adults with magnetic resonance (MR) diffusion tensor imaging (DTI).

METHODSDiffusion tensor images were obtained from 30 healthy volunteers without any cerebral abnormalities on conventional MRI. FA and the mean diffusivity (MD) of the optic radiation were measured in the directional encoded color (DEC) maps. The architecture of the optic radiation fiber tracts were displayed with the software of diffusion tensor fiber tracking.

RESULTSIn all subjects, the optic radiation could be readily identified in the DEC maps. The FA value was 0.509-/+0.029 in the left and 0.502-/+0.026 in the right, with the MD value of (0.763-/+0.050) x10(-3) and 0.748-/+0.052)x10(-3) mm2/s, respectively. No significant differences were found in the FA or MD value of the bilateral optic radiation (P>0.05). Diffusion tensor tractography (DTT) demonstrated that the 3 bundles of the optic radiation fibers were located in the lateral sagittal stratum, passing from the lateral geniculate body of the thalamus to the primary visual cortex. The dorsal and lateral bundles passed posteriorly to the superior bank of the calcarine cortex, while the ventral bundle passed anteriorly before making a sharp turn, known as the Meyer loop, and subsequently coursed posteriorly to terminate in the inferior margin of the calcarine cortex, which was consistent with the results of classic anatomical studies.

CONCLUSIONAs a novel method to study the relationship between visual function and optic pathway, DTI and DTT can show the FA and architecture of the optic radiation.

Adult ; Anisotropy ; Diffusion Magnetic Resonance Imaging ; methods ; Echo-Planar Imaging ; methods ; Female ; Geniculate Bodies ; anatomy & histology ; Humans ; Male ; Middle Aged ; Models, Anatomic ; Occipital Lobe ; anatomy & histology ; Optic Nerve ; anatomy & histology ; Visual Pathways ; anatomy & histology ; Young Adult

BACKGROUNDSmad4 is found mutated in many cancers. It acts as a tumor suppressor in the regulation of TGF-β signaling pathway. The objective of this work was to study the expression of Smad4 in intrahepatic cholangiocarcinoma (ICC) and its relationship with the biological behavior and prognosis of the disease.

METHODSForty-nine paraffin-embedded ICC specimens and nine normal liver tissues were analyzed by immunohistochemical methods using Smad4 monoclonal antibodies. The expression of Smad4 was compared with the clinical pathological characteristics of the patients.

RESULTSThe expression of Smad4 was 100% positive in normal liver tissues, which was higher than that in the ICC (44.9%). Negative labeling of the Smad4 protein was found in 26.1% (6/23) of well-differentiated ICCs and 61.5% (16/26) of poorly to moderately differentiated ICCs, and 34.3% (12/35) and 71.4% (10/14) showed negative Smad4 labeling (P = 0.018) of ICC at pathological Tumor Node Metastasis (pTNM) stage I-II and pTNM stage III-IV separately. Furthermore, 72% (8/11) of lymph node metastatic ICCs and 73.3% (11/15) of intrahepatic metastatic ICCs showed negative labeling of the Smad4 protein. The loss of Smad4 expression in those metastatic ICCs was significantly more severe compared with non-metastatic ICCs (P = 0.000).

CONCLUSIONSThe expression of Smad4 was associated with the histological grade, clinical stage, and metastasis of ICC (P < 0.05). The detection of Smad4 may be helpful in determining the degree of malignancy and prognosis of ICC.

Adult ; Aged ; Bile Duct Neoplasms ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; chemistry ; pathology ; Female ; Humans ; Liver Neoplasms ; chemistry ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Signal Transduction ; physiology ; Smad4 Protein ; analysis ; genetics ; physiology ; Transforming Growth Factor beta ; physiology

Objective To evaluate clinical significance of direct antiglobulin testing(DAT)in anemia in patients with severe chronic hepatitis B(CHB).Methods Red blood cell(RBC)-related parameters detection and DAT were performed among 30 healthy persons,30 CHB patients,and 50 severe CHB patients,clinical factors related to posi-tive DAT were analyzed.Results RBC count,hemoglobin (Hb)concentration,and hematocrit(HCT)level in severe CHB patients were all lower than CHB patients and healthy group(P <0.05),RBC distribution width(RDW)in severe CHB patients were all higher than CHB patients and healthy group(P<0.05);the positive rate of DAT in patients with se-vere CHB,CHB,and healthy group were 62.82%,13.33% and 0 respectively.RBC count,Hb concentration,and HCT level in severe CHB patients with positive DAT were all lower than severe CHB patients with negative DAT (all P <0.05),while RDW was higher than the latter (P=0.001);after RBC was separated through capillary,positive intensity of DAT of aged RBCs was higher than young RBCs in severe CHB patients (P <0.001);among severe CHB patients, DAT-positive and-negative patients differed in gender,age,alanine aminotransferase,total bilirubin,complement C3, C-reactive protein,and complication of diabetes(all P≤0.05).Conclusion Anemia in severe CHB patients may be re-lated to immune hemolysis of aged RBCs induced by antibody adsorption.

OBJECTIVETo study the combined effect of gestational age and birth weight on metabolites related to inherited metabolic diseases (IMD).

METHODSA total of 3 381 samples ruled out of IMD by follow-up were randomly selected from 38 931 newborns who participated in the neonatal IMD screening during 2014-2016. The 3 381 neonates were categorized into seven groups according to their gestational age and birth weight: extremely preterm appropriate-for-gestational age (AGA) group (n=12), preterm small-for-gestational age (SGA) group (n=18), preterm AGA group (n=219), preterm large-for-gestational age (LGA) group (n=18), full-term SGA group (n=206), full-term AGA group (n=2 677), and full-term LGA group (n=231). Heel blood samples were collected from each group on postnatal days 3-7 after adequate breastfeeding. Levels of 17 key IMD-related metabolic indices in dried blood spots were measured using tandem mass spectrometry. Spearman′s correlation analysis was used to investigate the relationships between 17 IMD-related metabolic indices and their influencing factors, while covariance analysis was used to compare the metabolic indices between these groups.

RESULTSAfter adjusting the influencing factors such as physiological and pathological status, compared with the full-term AGA group, the extremely preterm AGA, preterm SGA, and preterm AGA groups had significantly reduced levels of leucine\isoleucine\hydroxyproline and valine (P<0.05); the preterm AGA group had a significantly decreased ornithine level (P<0.05); the extremely preterm AGA and preterm AGA groups had a significantly reduced proline level (P<0.05). Besides, the phenylalanine level in the extremely preterm AGA and preterm AGA groups, the methionine level in the preterm SGA group, and the tyrosine level in the preterm AGA group all significantly increased (P<0.05). The increased levels of free carnitine, acetylcarnitine, and propionylcarnitine were found in the preterm SGA and preterm AGA groups. The oleylcarnitine level also significantly increased in the preterm SGA group (P<0.05). Most carnitine indices showed significant differences between the SGA group and the AGA/LGA group in both preterm and full-term infants (P<0.05).

CONCLUSIONSLow gestational age and low birth weight may result in abnormal results in IMD screening. Therefore, gestational age and birth weight should be considered to comprehensively judge the abnormal results in IMD screening.

Birth Weight ; Female ; Gestational Age ; Humans ; Infant, Newborn ; Infant, Small for Gestational Age ; Male ; Metabolic Diseases ; metabolism

Humans ; Norepinephrine ; Bacterial Infections/drug therapy* ; Immunity, Innate