[Effect of miR-23b on the malignant phenotype and the sensitivity of lenvatinib in human hepatocellular carcinoma cells].

Objective: To investigate the effect of miR-23b on the malignant phenotype and the sensitivity of lenvatinib in human hepatocellular carcinoma cells. Methods: Human hepatocellular carcinoma cell line HepG2, SMMC-7721 and QGY-7703 were transfected with miR-23b mimic and its control, respectively. CCK-8 and EdU assay were used to detect cell proliferation. Transwell assay were used to detect changes in cell migration and invasion. Tube formation assay were used to detect vasculogenic mimicry formation. The comparison of the mean between groups was analyzed by t-test. Results: CCK-8 results showed that the A values ​​of human hepatocellular carcinoma cell line HepG2 and SMMC-7721 in the miR-23b mimic group were 0.325 ± 0.011 and 0.537 ± 0.026, respectively, which were significantly lower than the control group 0.430±0.017 and 0.752 ± 0.051 (P < 0.05). Transwell assay result showed that the number of cell migration of human hepatocellular carcinoma cell line HepG2 and SMMC-7721 in the miR-23b mimic group was (517.220 ± 32.873) and (242.327 ± 20.793), respectively, which were significantly lower than that of the control group (724.130 ± 15.142) and (424.432 ± 27.212) (P < 0.01). Simultaneously, the number of cell invasions in the miR-23b mimic group were (55.671 ± 7.514) and (64.670 ± 6.011), respectively, which were significantly lower than those in the control group (124.320 ± 11.782) and (156.204 ± 12.501) (P < 0.01). Tube formation assay showed that the number of tube forming branches of hepatocellular carcinoma cell line QGY-7703 and SMMC-7721 in the miR-23b mimic group was (489.824 ± 42.035) and (435.201 ± 44.143), respectively, which were significantly lower than that of the control group (878.620 ± 31.618) and (785.430 ± 38.723) (P ​​< 0.01). In addition, EdU results showed that after miR-23b combined with lenvatinib, the positive rates of EdU staining of hepatocellular carcinoma cell line HepG2 and SMMC-7721 in the miR-23b mimic group were (32.905 ± 1.342)% and (24.811 ± 0.820)%, respectively, which were significantly lower than the control group (52.623 ± 2.441)% and (38.702 ± 1.312)% (P < 0.05). Conclusion: miR-23b can inhibit the proliferation, migration, invasion and vasculogenic mimicry formation, and enhance the sensitivity of lenvatinib drug in human hepatocellular carcinoma cells.

A morphometric study of the thoracolumbar spine spinous process and lamina space in the Chinese.

BACKGROUND: The aim of the study was to analyse the anatomical parameters of the thoracolumbar spine spinous process and lamina space for Chinese anatomic study, and provide an anatomical reference for its clinical operation. MATERIALS AND METHODS: Samples from 24 adult autopsy subjects were obtained from the thoracolumbar spine spinous process and lamina space at levels T1 through L5. Direct measurements were made for the spinous process spacing distance, spinous process length, width, thickness and inclination angle, and the lamina space width and height. RESULTS: 1. Distance of the spine spinous process spacing: Thoracic part: The maximum tip distance was observed at T4~T5 level, and the minimum tip distance was observed at T9~T10 level. The maximum centre distance and root distance were observed at T11~T12 level, and the minimum were observed at T5~T6 level separately. Lumbar part: distance of spinous process spacing in lumbar part showed a decreasing pattern from L1~L2 to L5~S1. 2. Length, width, thickness of the spine spinous process: 1) The length of the spinous process: The upper border gradually increased from T1 to T6 and then decreased till T12 region. The centre region is T8 maximum, T11 minimum. The lower border length showed a decreasing trend from T1 to T12. Lumbar part: The length increased from L1 and reached maximum value at L3. Then, the length decreased gradually to reach minimum value at L5. 2) The width of the spinous process: The width showed an increasing trend from T1 to T12. Lumbar part: Maximum width was seen at L3 and a minimum L5. 3) The thickness of the spinous process: Tip thickness > Centre thickness > Root thickness in each thoracic and lumbar vertebra. Thoracic part: the maximum tip thickness is T1, T7 minimum, The maximum centre thickness is T12, T7 minimum. The maximum root height is T6, T9 minimum. Lumbar part: Maximum tip thickness was seen at L1, and a minimum L3. Maximum centre thickness was seen at L5, and a minimum L2. Maximum root thickness was seen at L2, and a minimum L1. 3. Inclination angle of the spine spinous process: The inclination angle gradually decreased from T1 to T7 to minimum value at T7 and then increased till T12 region. 4. Width and height of lamina space: 1) The width of lamina space: For thoracic part, the data became shorter gradually from T1~T2 to T5~T6, and then increased till to T11~T12. For lumbar part, the width of lamina space increased from T12~L1 to L5~S1. 2) The height of lamina space: In the thoracic vertebrae, the maximum height of centre region was observed at T11~T12 and the minimum mean value was observed at T3~T4. In the lumbar vertebrae, the height of the lamina space was gradually increased from T12~L1 to L5~S1. CONCLUSIONS: This study reports morphometric data of the thoracolumbar spine spinous process and lamina space in the Chinese population, which provides an anatomic basis for thoracolumbar spine design of internal fixation, posterior surgery, puncture and epidural anaesthesia.

The involvement of protein kinases in the cardioprotective effect of chronic hypoxia.

The purpose of this review is to analyze the involvement of protein kinases in the cardioprotective mechanism induced by chronic hypoxia. It has been reported that chronic intermittent hypoxia contributes to increased expression of the following kinases in the myocardium: PKCdelta, PKCalpha, p-PKCepsilon, p-PKCalpha, AMPK, p-AMPK, CaMKII, p-ERK1/2, p-Akt, PI3-kinase, p-p38, HK-1, and HK-2; whereas, chronic normobaric hypoxia promotes increased expression of the following kinases in the myocardium: PKCepsilon, PKCbetaII, PKCeta, CaMKII, p-ERK1/2, p-Akt, p-p38, HK-1, and HK-2. However, CNH does not promote enhanced expression of the AMPK and JNK kinases. Adaptation to hypoxia enhances HK-2 association with mitochondria and causes translocation of PKCdelta, PKCbetaII, and PKCeta to the mitochondria. It has been shown that PKCdelta, PKCepsilon, ERK1/2, and MEK1/2 are involved in the cardioprotective effect of chronic hypoxia. The role of other kinases in the cardioprotective effect of adaptation to hypoxia requires further research.

[Establishment of nutrition literacy core items for Chinese preschool children].

Objective: To establish the nutrition literacy core items for preschool children in China. Methods: The framework system and preliminary items of nutrition literacy were established through literature review and experts consultation. E-Delphi method was used to determine the nutrition literacy items. Nine experts in the field of nutrition, child and adolescent health, health education were invited to score the importance of each nutrition literacy item via Email. The judgment basis and familiarity of experts towards the items, active coefficient, authority coefficient and harmony coefficient were evaluated and analyzed to generate the final list of nutrition literacy items. Results: In the first round of E-Delphi consultation, the active coefficient and authority coefficient were 88.9%(8/9) and(0.88±0.07).The harmony coefficient was 0.350 (χ2=36.432, P=0.001). In the second round, the active coefficient and authority coefficient were 100% (9/9) and (0.89±0.11).The harmony coefficient was 0.347 (χ2=34.330, P<0.001). After qualitative and quantitative analyses, we identified the final list of nutrition literacy items consisting of two scales (basic knowledge, lifestyles and dietary behaviors), six subscales (food knowledge, food characteristic, food choice, dietary behavior, dietary safety, and physical activity)with 14 items in total. Conclusion: The framework system and core items of nutrition literacy are established for Chinese preschool children, which could provide a reference basis for the implementation of nutrition education project and the establishment of standardized evaluation tools.

Non-Ohmic Variable-Range Hopping and Resistive Switching in SrTiO_{3} Domain Walls.

We report observation of electric field driven conductivity with negative differential conductance and resistive switching in insulating SrTiO_{3} samples over a wide range of applied voltages at low temperatures. The observed current follows I=I_{0}exp[-(E^{*}/E)^{1/2}] at large applied electric field, corresponding to variable range hopping conduction with a Coulomb gap in domain walls. Our data are sufficient to discriminate unambiguously between Shklovskii and Mott hopping via their different electric field exponent. Under some conditions space-charge-limited currents are observed, and the charge mobility limit is determined to be in the range of 17 and 210 cm^{2}/Vs.

Resveratrol ameliorates high-fat diet-induced insulin resistance and fatty acid oxidation via ATM-AMPK axis in skeletal muscle.

OBJECTIVE: Resveratrol (RSV) is a polyphenolic phytoalexin that exhibits diverse pharmacological actions, including its effect on the insulin resistance. However, the mechanism through which RSV improves insulin resistance is not fully understood yet. The aim of this study was to determine the mechanism through which RSV ameliorates insulin resistance in skeletal muscle of high-fat diet (HFD)-induced mouse model, as well as palmitic acid (PA) treated L6 cells, with a specific focus on the response of RSV on fatty acid oxidation. MATERIALS AND METHODS: Male C57BL6/J mice were randomly divided into three groups: normal diet-fed mice (ND), the high-fat diet-fed mice (HFD), HFD supplemented with RSV (100 mg/kg body weight [BW]/day orally; n = 10). Fasting plasma glucose, insulin, total cholesterol, triglyceride (TG), and free fatty acid levels were determined. The intraperitoneal glucose tolerance test was used to measure blood glucose and area under the curve. The quantitative insulin sensitivity index was calculated to assess insulin resistance. Skeletal muscles were collected for histology study and protein expression measurement. L6 cells were cultured with PA and the glucose concentration in the culture medium, and the intracellular TG levels were tested. RSV, chloroquine, palmitoyltransferase and Ku-55933 were administered to differentiate L6 cells. RESULTS: The HFD fed mice showed increased BW, hyperglycemia, and hyperlipidemia. The expressions of ataxia telangiectasia mutated (ATM), 5' adenosine monophosphate-activated protein kinase (AMPK), carnitine palmitoyltransferase 1, cytochrome oxidase subunit IV protein were significantly decreased in the skeletal muscles of HFD fed mice and PA-treated L6 cells. All these effects induced by HFD and PA were reversed by RSV treatment. CONCLUSIONS: ATM is a key factor to improve HFD-induced lipid metabolism and insulin resistance in skeletal muscles. The effects of RSV on ameliorating HFD-induced abnormal lipid metabolism and insulin resistance mediated through ATM-AMPK pathway may due to its improvement in fatty acid oxidation efficiency and sequential reduction in ROS production in skeletal muscle. These results provide important theoretical evidence for the application of RSV in the prevention and treatment of diabetes mellitus and related metabolic diseases.

Determination of the Content of 4-FMA in Rat Plasma Samples by HPLC-MS/MS Method.

ABSTRACT: Objective To develop a high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method for the determination of the content of 4-fluoromethamphetamine （4-FMA） in rat plasma, and to provide a methodological basis for the study of the toxicokinetics of 4-FMA in rats. Methods Rat plasma samples were added into internal standard methamphetamine （MA）. Its proteins were precipitated with methanol and then separated with Poroshell 120 EC-C18 chromatographic column. A 0.1% formic acid aqueous solution and a 0.1% formic acid acetonitrile solution were used as the mobile phase at the flow rate of 0.4 mL/min. Electrospray ionization source was used for detection in the multiple reaction monitoring （MRM） mode. Results The linear relationship was good when the mass concentration of 4-FMA in plasma samples was in the range of 5-1 000 ng/mL （r>0.999）. The limit of detection （LOD） was 3 ng/mL and the limit of quantification （LOQ） was 5 ng/mL. The accuracy was expressed as relative error （RE）, and in the range of ±5%, the intra-day precision and inter-day precision （relative standard deviation, RSD） less than 9%, and the extraction recovery rate was more than 90%. The analysis and detection of plasma samples were completed within 2.5 min. Conclusion This study developed a HPLC-MS/MS method for the determination of 4-FMA in rat plasma samples. This method is accurate, rapid, simple and sensitive and can be applied to the study of toxicokinetics of 4-FMA.

[An analysis of relative factors, bacteriological profile and antibiogram of neonatal sepsis].

Objective: This study was designed to identify the relative factors, bacteriological profile and their antibiotic susceptibility pattern of neonatal sepsis. Methods: A retrospective survey was conducted on the clinical information, pathogen identification and antibiotic sensitivity results of 425 newborns with neonatal sepsis admitted to Nanjing Maternity and Child Health Care Hospital from 2010 to 2017. Of the 425 positive blood-cultures, 148 (34.82%) were early-onset neonatal sepsis (EOS) and 277 (65.18%) were late-onset neonatal sepsis (LOS). Clinical information and pathogen identification were compared between EOS and LOS. Antibiotic sensitivity of gram negative organisms (G(-)) and gram positive organisms (G(+)) were also detected. Results: The rates of premature delivery (78.70%, n=218), low birth weights (67.15%, n=186) and cesarean delivery (59.57%, n=165) were significantly increased in LOS (P<0.05) compared with those rates in EOS, which were 41.89% (n=62), 37.84% (n=56) and 46.62% (n=69). Parturients fever (18.24%, n=27) and meconium like amniotic fluid (25.68%, n=38) were significantly increased in EOS (P<0.05) compared with those rates in LOS, which were 7.94% (n=22) and 5.42% (n=15). Among the identified pathogen, the incidence of G(-) and G(+) bacteria were 216 (50.83%) and 201 (47.29%) respectively, and the rest was Candida glabrata (1.88%, n=8). Escherichia coli 68 (16.00%) was the predominant isolate followed by Klebsiella pneumoniae (13.18%, n=56). The detection rate of Hemolytic staphylococcus (10.81%, n=16) was significantly increased in EOS (P<0.001) compared with LOS (1.44%, n=4). However, the incidence of Klebsiella pneumonia (5.88%, n=44) was higher in LOS (P=0.024) compared with EOS (8.11%, n=12). Most of the gram positive isolates exhibited high resistance to penicillin (90.32%-100.00%) and cephalosporin group antibiotics (25.00%-100.00%). Similarly, the majority of the gram negative isolates showed higher resistance to ampicillin (83.33%-100.00%), but susceptible to aminoglycosides (0-11.76%), quinolones (0-17.65%) and ß-lactams (0-5.88%). Conclusion: Among the study population, the percent of preterm, low birth weight and cesarean section were higher in LOS while parturients fever and meconium-like amniotic fluid were higher in EOS. The pathogens with the highest detection rate were Escherichia coli and Klebsiella pneumoniae. The results of antibiotic susceptibility test showed that common pathogens had high resistance to commonly used antibacterial drugs.

FHL2 participates in renal interstitial fibrosis by altering the phenotype of renal tubular epithelial cells via regulating the ß-catenin pathway.

OBJECTIVE: To investigate the potential role of FHL2 (four and a half LIM domains protein 2) in the renal interstitial fibrosis and its underlying mechanism. MATERIALS AND METHODS: NRK-52E, the rat tubular epithelial cell line, was selected for the in vitro experiments. The unilateral ureteral obstruction (UUO) mouse model and phenotype changes of NRK-52E cells were induced by TGF-ß (transforming growth factor-ß) treatment. Protein and mRNA expressions of biomarkers of tubular cells and renal fibrosis in NRK-52E cells were detected. Meanwhile, phenotype changes of NRK-52E cells were detected after FHL2 overexpression. Furthermore, CD1 mice were selected for constructing the UUO mouse model. Protein and mRNA expressions of biomarkers of tubular cells and renal fibrosis in kidney tissues were detected. CD1 mice with FHL2 overexpression were constructed by tail vein injection of FHL2 plasmid for further observation of renal interstitial fibrosis. Expressions of ß-catenin pathway-related genes were detected by Western blot and polymerase chain reaction (PCR), respectively. RESULTS: The FHL2 expression was increased during the phenotype change of NRK-52E cells induced by TGF-ß treatment. Overexpression of FHL2 led to a significant phenotype change. Similarly, the FHL2 expression was elevated in the UUO mouse model. Renal interstitial fibrosis was exaggerated and expression levels of genes related to the ß-catenin pathway were increased after injection of FHL2 plasmid. CONCLUSIONS: FHL2 is involved in renal interstitial fibrosis by altering the phenotype of renal tubular epithelial cells via regulating the ß-catenin pathway.

Silencing of LncRNA TCONS_00088786 reduces renal fibrosis through miR-132.

OBJECTIVE: To examine the role of long non-coding ribonucleic acid (LncRNA) TCONS_00088786 in the development of renal interstitial fibrosis and its potential mechanism in this process. MATERIALS AND METHODS: Unilateral ureteral obstruction (UUO) was used to induce tubulointerstitial fibrosis. Masson staining showed the degree of renal fibrosis in UUO mice. Immunohistochemistry and immunofluorescence were performed to detect the fibrosis-related proteins, the 24-h urine volume and protein content. The renal functions were reflected via serum creatinine (Scr) and blood urea nitrogen (BUN). Changes in lncRNATCONS_00088786, miR-132 and collagen I and III in the development process of renal fibrosis were detected through reverse transcription-polymerase chain reaction (RT-PCR). Small interfering RNA (siRNA) was transfected into NRK52E cells to mimic the knockdown. Western blot was adopted to detect the changes in miR-132, collagen I and III after the siRNA was transfected by transforming growth factor-ß (TGF-ß) for 24 h. RESULTS: With the development of renal fibrosis, lncRNA TCONS_00088786 and miR-132 were increased gradually. After the knockdown of lncRNA TCONS_00088786, miR-132 was decreased and fibrosis-related protein was also decreased. CONCLUSIONS: Decreased lncRNA TCONS _00088786 inhibits renal interstitial fibrosis by reducing miR-132 and it may be a potential novel molecular target for the treatment of renal interstitial fibrosis.

Local Electrical Imaging of Tetragonal Domains and Field-Induced Ferroelectric Twin Walls in Conducting SrTiO_{3}.

We demonstrate electrical mapping of tetragonal domains and electric field-induced twin walls in SrTiO_{3} as a function of temperature and gate bias utilizing the conducting LaAlO_{3}/SrTiO_{3} interface and low-temperature scanning electron microscopy. Conducting twin walls appear below 105 K, and new twin patterns are observed after thermal cycling through the transition or on electric field gating. The nature of the twin walls is confirmed by calculating their intersection angles for different substrate orientations. Numerous walls formed when a large side- or back-gate voltage is applied are identified as field-induced ferroelectric twin walls in the paraelectric tetragonal matrix. The walls persist after switching off the electric field and on thermal cycling below 105 K. These observations point to a new type of ferroelectric functionality in SrTiO_{3}, which could be exploited together with magnetism and superconductivity in a multifunctional context.

Aspirin-triggered Lipoxin A4 attenuates mechanical allodynia in association with inhibiting spinal JAK2/STAT3 signaling in neuropathic pain in rats.

Aspirin-triggered Lipoxin A4 (ATL), as a Lipoxin A4 (LXA4) epimer, is endogenously produced by aspirin-acetylated cycloxygenase-2 (COX-2) and plays a vital role in endogenous anti-inflammation via the LXA4 receptor (ALX). Recent investigations have indicated that spinal neuroinflammation and the activation of the Janus Kinase 2 (JAK2)/Signal Transducers and Transcription Activators 3 (STAT3) signaling pathway are involved in neuropathic pain states. However, the effect of ATL on neuroinflammation and JAK2/STAT3 signaling in chronic constriction injury (CCI)-induced neuropathic pain in rats has not been well-studied. The present study demonstrated the anti-inflammatory and analgesic effect of ATL on neuropathic pain and assessed the role of spinal JAK2/STAT3 signaling on the effect of ATL. Intrathecal administration of ATL significantly attenuated mechanical allodynia via spinal ALX and inhibited the upregulation of pro-inflammatory cytokines (IL-1ß, IL-6, and TNF-α) on day 7 of CCI surgery. In addition, ATL markedly suppressed the upregulation of p-STAT3 induced by the neuropathic pain. Blockade of JAK2-STAT3 signaling with intrathecal administration of the JAK2 inhibitor AG490 or the STAT3 inhibitor S3I-201 clearly reduced mechanical allodynia and the upregulation of pro-inflammatory cytokines in CCI rats. Interestingly, inhibition of JAK2/STAT3 signaling via ATL or the specific signaling inhibitor (AG49, S3I-201) further promoted the increased expression of suppressor of cytokine signaling 3 (SOCS3) mRNA in the spinal cord induced by CCI surgery. Taken together, our results suggested that the analgesic effect of ATL was mediated by inhibiting spinal JAK2/STAT3 signaling and hence the spinal neuroinflammation in CCI rats.

Involvement of the spinal NALP1 inflammasome in neuropathic pain and aspirin-triggered-15-epi-lipoxin A4 induced analgesia.

Neuroinflammation plays an important role in nerve-injury-induced neuropathic pain, but the explicit molecular mechanisms of neuroinflammation in neuropathic pain remain unclear. As one of the most critical inflammatory cytokines, interleukin-1ß (IL-1ß) has been regarded as broadly involved in the pathology of neuropathic pain. The inflammasome caspase-1 platform is one primary mechanism responsible for the maturation of IL-1ß. Lipoxins, a type of endogenous anti-inflammatory lipid, have proved to be effective in relieving neuropathic pain behaviors. The present study was designed to examine whether the inflammasome caspase-1 IL-1ß platform is involved in chronic constriction injury (CCI)-induced neuropathic pain and in lipoxin-induced analgesia. After rats were subjected to the CCI surgery, mature IL-1ß was significantly increased in the ipsilateral spinal cord, and the inflammasome platform consisting of NALP1 (NAcht leucine-rich-repeat protein 1), caspase-1 and ASC (apoptosis-associated speck-like protein containing a caspase-activating recruitment domain) was also activated in spinal astrocytes and neurons, especially at the superficial laminae of the spinal dorsal horn; The aspirin-triggered-15-epi-lipoxin A4 (ATL), which shares the potent actions of the endogenous lipoxins, was administered to the CCI rats. Repeated intrathecal injection with ATL markedly attenuated the CCI-induced thermal hyperalgesia and significantly inhibited NALP1 inflammasome activation, caspase-1 cleavage, and IL-1ß maturation. These results suggested that spinal NALP1 inflammasome was involved in the CCI-induced neuropathic pain and that the analgesic effect of ATL was associated with suppressing NALP1 inflammasome activation.

Intention to comply with post-exposure management among nurses exposed to blood and body fluids in Taiwan: application of the theory of planned behaviour.

Nurses are at significant risk from occupationally acquired bloodborne virus infections following a needlestick and sharps injury. This study aimed to apply the theory of planned behaviour (TPB) to predict nurses' intention to comply with occupational post-exposure management. A cross-sectional survey was applied to select registered nurses who worked in human immunodeficiency virus (HIV)-designated hospitals. An anonymous, self-administered questionnaire based on the TPB was distributed to 1630 nurses and 1134 (69.5%) questionnaires were returned. From these, a total of 802 nurses (71%) reported blood and body fluid exposure incidents during 2003-2005 and this group was used for analysis. Only 44.6% of the 121 exposed nurses who were prescribed post-exposure prophylaxis (PEP) by infectious disease doctors returned to the clinic for interim monitoring, and only 56.6% of exposed nurses confirmed their final serology status. Structural equation modelling was used to test the TPB indicating perceived behavioural control (the perception of the difficulty or ease of PEP management, ß=0.58), subjective norm (the perception of social pressure to adhere to PEP, ß=0.15), and attitudes (ß=0.12) were significant direct effects on nurses' intention to comply with post-exposure management. The hypothesised model test indicated that the model fitted with the expected relationships and directions of theoretical constructs [χ(2) (14, N=802)=23.14, P=0.057, GFI=0.987, RMSEA=0.039]. The TPB model constructs accounted for 54% of the variance in nurses' intention to comply with post-exposure management. The TPB is an appropriate model for predicting nurses' intention to comply with post-exposure management. Healthcare facilities should have policies to decrease the inconvenience of follow-up to encourage nurses to comply with post-exposure management.

Treating sewage using coimmobilized system of Chlorella pyrenoidosa and activated sludge.

Chlorella pyrenoidosa was coimmobilized with activated sludge to produce algae-bacteria beads for sewage treatment. Hydrolysis/acidogenesis pretreatment could improve the symbiotic microenvironment of coimmobilized Chlorella pyrenoidosa and activated sludge, and as a result, promote the removal of nutrients (COD(cr), inorganic nitrogen and inorganic phosphorus) in the sewage. A photo-bioreactor combining hydrolysis/acidogenesis pretreatment and coimmobilized technique was designed to treat sewage continuously. The results show that, the removal efficiencies of COD(cr), NH4(+)-N and TP reached steady state after 4-days of experiment. The removal efficiencies of COD(cr), NH4(+)-N and TP were 59.6%, 59.0% and 60.3% respectively.

Diversity of human leukocyte antigen-B27 alleles in Han population of Hunan province, southern China.

This study was to investigate the frequency of HLA-B27 and its subtypes in the Han population of Hunan province, southern China. One hundred and sixty-nine healthy unrelated donors were tested for HLA-B27 by polymerase chain reaction-sequence-specific primer (PCR-SSP). One hundred and twenty-eight B27-positive spondyloarthropathy patients and 18 B27-positive healthy controls were subtyped using the high-resolution PCR-SSP. The phenotype frequency of human leukocyte antigen (HLA)-B27 was found to be 2.36% in healthy population. Five B27 alleles were identified: B*2704, B*2705, B*2706, B*2707, and B*2724. No significant difference was found in the distribution of HLA-B27 subtypes between the patients and controls studied. Notably, B*2724 was observed in a juvenile patient with ankylosing spondylitis. This subtype has not been previously reported in Chinese ankylosing spondylitis (AS) patients and other ethnic groups.

Generalized granuloma annulare associated with chronic hepatitis B virus infection.

We describe a 63-year-old Chinese woman with generalized granuloma annulare (GGA) associated with chronic hepatitis B virus (HBV) infection for 10 years. She developed non-annular papule lesions after a previous infection of hepatitis B virus. Since then, her clinical course was aggressive with involvement of the trunk, extremities and neck. Histologically, granulomatous inflammations were found in the upper to mid dermis. The HBV DNA in these lesions was demonstrated by polymerase chain reaction (PCR). GGA was totally regressed after treatment of interferon-alpha for 3 months. To the best of our knowledge, GGA associated with chronic HBV infection has never been reported in literature. The prognostic significance of the association of granulomatous inflammation and virus infection is reviewed.

Human herpesvirus 8 open reading frame 26 and open reading frame 65 sequences from multiple myeloma patients: a shared pattern not found in Kaposi's sarcoma or primary effusion lymphoma.

Human herpesvirus 8 (HHV-8), also known as Kaposi's sarcoma-associated herpesvirus, has been implicated in the pathogenesis of Kaposi's sarcoma (KS), primary effusion lymphoma (PEL), multicentric Castleman's disease, and recently multiple myeloma (MM). DNA sequence analyses of HHV-8 suggest that multiple HHV-8 strains exist. We extracted DNA from 24 patients with MM and 3 patients with monoclonal gammopathy of undetermined significance and compared HHV-8 open reading frames (ORFs) 26 and 65 sequences with those derived from patients with KS, PEL, and two HHV-8-positive PEL cell lines KS-1 and BC-1. ORF26 sequence data suggest that MM patients are consistently carriers of HHV-8 strain subtype C3. All MM patients also consistently revealed either a single bp deletion or substitution at position 112197 in ORF65. This unique alteration is not present in patients with KS or PEL or in PEL cell lines. It occurs in the portion of ORF65 that is known to be responsible for a serological response to HHV-8.

Cloning of the first invertebrate MAGE paralogue: an epitope that activates T-cells in humans is highly conserved in evolution.

The MAGE (Melanoma Associated Antigen) family tumor-specific antigens are shared by a number of histologically different tumors. Till date, only human and mouse MAGE genes have been characterized. Our study describes the first non-mammalian member of MAGE super-family, DMAGE from D. melanogaster. A conceptual translation of the cDNA of DMAGE identifies a putative protein that contains a motif that shares eight out of nine amino acids with the previously identified promiscuous, HLA-A2 restricted antigenic epitope in the C-terminus of human MAGE-B1 and -B2. Similarly, this motif of DMAGE shares seven out of nine amino acids with the same antigenic epitope of human MAGE-A3 and -A12. Thus, the phylogeny of proteins that activate tumor specific T-cells in mammals as unmutated self-proteins began at least 100 million years earlier in evolution than the emergence of the adaptive immune system of higher vertebrates. Northern analysis revealed that DMAGE is a developmentally regulated gene highly expressed in adult fruit fly and in the embryo of D. melanogaster. In contrast, the expression level of the mRNA of DMAGE in fruit fly larva is substantially lower than in embryo and adult fly. We propose that studies of DMAGE on D. melanogaster may help define the function(s) of MAGE super-family genes.

[Transformation of sainfoin by Agrobacterium rhizogenes LBA9402 Bin19 and regeneration of transgenic plants].

Hypocotyl segments of Onobrychis viciaefolia were transformed by Agrobacterium rhizogenes LBA9402 which harboured pBin19 and pRi1855. Seedling age and preculture time of hypocotyl segments influenced the transformation frequency. Paper electrophoresis revealed that 70% of single hairy root cultures could synthesize agropine. Calli were induced from hairy root segments on MS medium containing 0-9.05 mumol/L 2,4-D and 0-2.22 mumol/L 6-BA at first, then they were transferred onto MS0 medium without kanamycin for regeneration. Constitution and concentration of phytohormones in callus induction media affected subsequent regeneration of calluses on MS0 medium remarkably. Regeneration frequency and shoot number per callus declined when 2,4-D concentration in callus induction media increased from 4.52 to 9.05 mumol/L, while they ascended when 6-BA in callus induction media increased from 0 to 2.22 mumol/L. On MS medium supplemented with 4.52 mumol/L 2,4-D and 2.22 mumol/L 6-BA, only 14.2% hairy root segments could produce calluses, but the regeneration frequency reached 58.1% and the shoot number per callus was 37.2. In 32 analysed plants regenerated from 8 kanamycin resistant hairy root lines, 25 were nptII positive and showed different copy numbers.