Dramatic Response to Pyrotinib and T-DM1 in HER2-Negative Metastatic Breast Cancer With 2 Activating HER2 Mutations.

HER2 signaling is activated in response to somatic HER2 mutations, which are often found in invasive lobular breast cancer (ILC) and are associated with poor prognosis. Tyrosine kinase inhibitors (TKIs) have demonstrated considerable antitumor activity in patients with HER2-mutated advanced breast cancer (BC). Further, several clinical trials have indicated that HER2-targeted antibody-drug conjugates (ADCs) exhibit promising efficacy in lung cancer with HER2 mutations, and the efficacy of ADCs against HER2-mutated BC is currently being evaluated. Several preclinical studies have demonstrated that the therapeutic efficacy of ADCs in HER2-mutated cancer can be enhanced by the addition of irreversible TKIs, but the potential of such a combined treatment regimen for the treatment of HER2-mutated BC has not been reported. Herein, we describe a case in which a patient with estrogen receptor-positive/HER2-negative metastatic ILC with 2 activating HER2 mutations (D769H and V777L) exhibited a significant and durable response to anti-HER2 treatment with pyrotinib (an irreversible TKI) in combination with ado-trastuzumab emtansine, which was administered after multiple lines of therapy that had resulted in disease progression. Further, based on the evidence from the present case, TKI plus ADC seems to be a promising combination anti-HER2 regimen for patients with HER2-negative/HER2-mutated advanced BC, although further rigorous studies are warranted to confirm these findings.

Occurrence of Nigrospora osmanthi Causing Leaf Blight on Water Lettuce in China.

Water lettuce (Pistia stratiotes L.), is one of the emerging invasive weeds for inland water bodies in Asia and become a major problem for local water ecosystem. Biocontrol of water lettuce by mycobiota is being considered as a promising and sustainable method (Kongjornrak et al. 2019). During July 2021, a leaf blight of water lettuce was observed within about 1.5 ha in Shenxi stream (N25°66', E119°05') in Putian, Fujian, China. The disease severity was about 100% with 80% incidence, early symptoms appeared as small irregularly yellow or brown blight, severely infected leaves turned to be rot, then death and sink. Small pieces (5 × 5 mm) of symptomatic leaves were excised and surface disinfected with 75% ethanol and 0.1% HgCl2 solution, air dried and plated on potato dextrose agar (PDA). 3~5 days after incubation at 28°C, six fungal pure cultures showing similar morphology were obtained from the infected leaves. On PDA, colonies were flat, aerial mycelium grew sparsely, most of it grew inside the agar medium, it reverses white to grey to black with age. Hyphae were branched, septate, smooth and hyaline. Conidiophores mostly reduced to conidiogenous cells and setae were not observed. Conidiogenous cells were monoblastic, discrete and solitary, at first hyaline, subspherical, then turning to pale brown, ampulliform, 4.5-10 × 3.5-6 µm in size. Conidia were solitary, globose or ellipsoidal, black, smooth, some of it formed directly from the mycelia, aseptate, 8-12 µm diam (n=10). Genomic DNA was extracted from one of the representative isolate Z1. ITS1/ITS4 (Mills et al. 1992), Bt-2a/Bt-2b (Glass and Donaldson 1995) and EF1-728F/EF-2 (O'Donnell et al. 1998) primer pairs were used to amplify the isolate's internal transcribed spacer (ITS), the Beta-tubulin fragment (TUB) and the partial translation elongation factor (TEF1), respectively. The isolate's sequences were deposited in the GenBank with accession numbers of OM279539 (ITS), OM296034 (TUB) and OM296035 (TEF1). Phylogenetic analysis using maximum likelihood based on the ITS-TUB-TEF1 concatenated sequences from Nigrospora species revealed that isolate Z1 is closely clustered with N. osmanthi strain LC4487. The fungus was identified as N. osmanthi based on the morphological characteristics and molecular analyses (Hao et al. 2020; Wang et al. 2017). Pathogenicity test were performed using twenty inoculated and control plants, respectively. Conidial suspensions (107 CFU/ml) of Z1 isolate were spray-inoculated on the leaves of healthy water lettuce seedlings, while sterile distilled water was used as control. Inoculated and control plants were kept in the differential 50-liter plastic tanks and maintained in a greenhouse at room temperature (19 to 24°C) for one month. Symptoms appeared 7 days post inoculation, which was similar to what occurs in the field. No symptoms occurred on controls. Pathogen was reisolated and confirmed by morphology and molecular analysis. Koch's postulates were conducted twice. N. osmanthi is a pathogenic fungus of many crop plants, such as buckwheat (Shen et al 2021), Java tea (Ismail et al. 2022) or buffalograss (Mei et al. 2019) in Asia and particularly in China. However, to our knowledge, this is the first report of N. osmanthi causing leaf blight on water lettuce. Further studies on how to apply formulated N. osmanthi will be required so that the strain could be effectively used to control water lettuce, moreover, its environmental safety also need a rigorous experimental evaluation.

The complete chloroplast genome of Polygala subopposita, an endemic milkwort in China.

Polygala subopposita is an endemic milkwort species in China. In this study, we present the assembly of its chloroplast genome (plastome) for the first time. The total plastome size is 164,784 bp in length, consisting of a large single-copy (LSC) region of 83,235 bp, a small single-copy (SSC) region of 8,037 bp, and two inverted repeat (IR) regions of 36,756 bp that have expanded approximately 10 kb into the SSC region. A total of 111 unique genes were identified in the plastome, including 77 protein-coding, 30 tRNA, and four rRNA genes. Interestingly, the trnQUUG gene was found to have two additional copies in the IRs, and the clpP gene lost its entire intron 2. Phylogenetic analysis suggests a close relationship between P. subopposita and P. crotalarioides. These findings provide valuable genomic resources for further research on the phylogenetic and evolutionary studies of Polygalaceae.

Safety and feasibility assessment of extending the flushing interval in totally implantable venous access port flushing during the non-treatment stage for patients with breast cancer.

Aim: To investigate the safety and feasibility of extending the flushing interval for the totally implantable venous access port (TIVAP) during the non-treatment stage in patients with breast cancer (BC) by retrospectively analyzing the patients' clinical data, including the incidence of TIVAP-related complications. Methods: This single-center retrospective study included patients with BC who underwent TIVAP implantation at our hospital between January 2018 and March 2021 during their non-treatment phase and visited the hospital regularly for TIVAP flushing. Among the 1013 patients with BC who received TIVAP implantation, 617 patients were finally included on the basis of the inclusion and exclusion criteria and divided into three groups according to the length of the flushing interval: group 1 (≤30 days, n = 79), group 2 (31-90 days, n = 66), and group 3 (91-120 days, n = 472). The basic characteristics of patients in each group and the incidence of TIVAP-related complications (catheter obstruction, infection, and thrombosis) were analyzed. Results: No significant intergroup differences were observed in age, body mass index (BMI), tumor stage, pathological staging, implantation approach, chemotherapy regimen, duration of treatment, and TIVAP-related blood return rate (P > 0.05). Among patients from all three groups, 11 cases of catheter pump-back without blood and eight cases of TIVAP-related complications such as infection, thrombosis, and catheter obstruction were recorded. However, no significant differences in TIVAP-related complications were observed among the three groups (P > 0.05). Conclusion: Extending the TIVAP flushing interval beyond three months during the non-treatment stage in BC patients is safe and feasible and did not increase the incidence of TIVAP-related complications.

Detection of Escherichia coli, Pseudomonas aeruginosa, Salmonella paratyphoid B, and Shigella dysentery in live Bacillus licheniformis products using propidium monoazide-real-time-quantitative polymerase chain reaction.

To eliminate the influences of excipients and interference of dead bacterial DNA on the detection of Escherichia coli, Pseudomonas aeruginosa, Salmonella paratyphoid B, and Shigella dysentery in live Bacillus licheniformis capsules, a polymerase chain reaction (PCR) method with high sensitivity and specificity was established. By combining bromide with propidium monoazide (PMA) -real-time quantitative PCR (qPCR) with microporous membrane filtration, excipients were removed, the filtrate was collected, and the bacteria were enriched using the centrifugal method. The optimal PMA working concentration, dark incubation time, and exposure time were determined. Specific E. coli, P. aeruginosa, S. paratyphoid B, and S. dysentery primers were selected to design different probes and a multiplex qPCR reaction system was established. The PMA-qPCR method was verified using different concentrations of dead and live bacteria. This method is efficient and accurate and can be widely applied to the detection of aforementioned pathogenic bacterial strains in live Bacillus licheniformis products.

Synthesis of bimetallic nickel cobalt selenide particles for high-performance hybrid supercapacitors.

Supercapacitors are known as promising excellent electrochemical energy storage devices because of their attractive features, including quick charge and discharge, high power density, low cost and high security. In this work, a series of litchi-like Ni-Co selenide particles were synthesized via a simple solvothermal method, and the Ni-Co compositions were carefully optimized to tune the charge storage performance, charge storage kinetics, and conductivity for battery-like supercapacitors. Interestingly, the optimal sample Ni0.95Co2.05Se4 exhibits a high capacity of 1038.75 F g-1 at 1 A g-1 and superior rate performance (retains 97.8% of the original capacity at 4 A g-1). Moreover, an asymmetric supercapacitor device was assembled based on the Ni0.95Co2.05Se4 cathode and activated carbon anode. The device of Ni0.95Co2.05Se4//active carbon (AC) reveals a peak energy density of 37.22 W h kg-1, and the corresponding peak power density reaches 800.90 W kg-1. This work provides a facile and effective way to synthesize transition metal selenides as high-performance supercapacitor electrode materials.

First Report of Cercospora rodmanii Causing Leaf and Petiole Lesions on Water Hyacinth in China.

The rapid spread and colonization of water hyacinth (Eichhornia crassipes) leads to a series of serious environmental problems for water bodies, prompting microbiologists to develop effective mycoherbicides to alleviate the water hyacinth population (Julien et al. 2001). In September 2020, numerous leaftip diebacks and petiole rots of water hyacinth, with 40 to 50% incidence, were observed within an area of 2 ha (ca. 2 km) mat on Jinjiang River tributary, Fujian, China. Ten infected leaf samples were collected and symptomatic tissues were cut into small pieces, surface disinfected in 75% ethanol followed by 0.1% MgCl2 solution and placed on potato dextrose agar (PDA). Pure cultures (Isolates J1 and J5) were obtained and their colonies on PDA appeared as white villi with wrinkled surfaces and dense colorless mycelium on the upper surface, while they were dark olivaceous-gray at the bottom. Internal mycelium consisted of septate, branched, smooth hyphae. There lacked stromata. Conidiophores were solitary to 2 to 5 in loose fascicles, thick-walled, conically truncated, 40 to 80 × 5 to 8 µm. Conidiogenous cells integrated, terminal, 30 to 150 × 4 to 6 µm, slightly protuberant, apical and lateral. Conidia were solitary, hyaline, acicular to obclavate, slightly curved, acute at the apex, 50 to 80 ×3 to 6 µm, indistinctly 1 to 3 septate. Genomic DNA from the two isolates was extracted for PCR, and the internal transcribed spacers (ITS), calmodulin (CAL), translation elongation factor 1-alpha (TEF), actin (ACT), histone H3 (H3) and chitin synthase (CHS) were amplified and sequenced using the primer pairs ITS1/ITS4, CL1/CL2A, EF1Fd/EF1Rd, ACT1Fd/ACT1Rd, CYLH3F/CYLH3R and CHS-79F/CHS-345R (Weir et al. 2012; White et al. 1990), respectively. The ITS (MZ436974-MZ436975), CAL (MZ519385-MZ519386), TEF (OK340826-OK340827), ACT (OK340824-OK340825), H3 (OK340828-OK340829) and CHS (MZ519387-MZ519388) sequences were deposited in GenBank. A phylogenetic tree based on concatenated sequences of ITS-CAL-TEF-ACT-H3-CHS from the genus Cercospora was constructed using a maximum likelihood method, showing that the present isolates and Cercospora rodmanii formed a monophyletic group with 99% bootstrap support. Therefore, the fungus was identified as C. rodmanii (Groenewald et al. 2013; Nguanhom et al. 2015). To test Koch's postulates, petioles of a set of 20 water hyacinth seedlings of 30 to 40 days old were wounded using a sterile needle and then spray-inoculated with 20 µl of the spore suspension of each isolate at 106 CFU/ml. Another set of 20 seedlings inoculated with sterile distilled water served as the controls. Inoculated plants were kept in 50-liter plastic tanks and maintained in a greenhouse at room temperature 22 to 28°C with a relative humidity of 80 to 85%. After 7 to 20 days, lesions were observed on the inoculated leaves but not on the control leaves. The same fungus was reisolated and identified by microscopy and PCR-sequencing. The pathogenicity test was conducted twice. Cercospora rodmanii and C. piaropi have been reported on water hyacinth in America, Brazil, México, and Zambia (Charudattan et al. 1985; Montenegro-Calderón, 2011; Moran, 2015). To our knowledge, this is the first report of C. rodmanii causing leaf and petiole lesions on water hyacinth in China. This report will help identify indigenous plant pathogens in China and develop a novel bioherbicide strategy for control of water hyacinth.

The complete chloroplast genome of Ormosia nuda (fabaceae), an endemic species in China.

Ormosia nuda is a legume species endemic to China. The chloroplast genome (plastome) of this species was assembled in this study. The total plastome size is 173,789 bp in length, containing a large single-copy (LSC) region of 73,847 bp, a small single-copy (SSC) region of 18,744 bp, and two inverted repeat (IR) regions of 40,599 bp which have expanded about 15 kb into LSC. The plastome encodes a total of 111 unique genes, including 77 protein-coding, 30 tRNA, and 4 rRNA genes. Phylogenetic analysis well resolved that O. nuda clustered with O. xylocarpa and O. emarginata. The plastome of O. nuda will provide informative genomic resources for further phylogenetic studies.

Mesoporous anion-cation-codoped Co9S8 nanorings for enhanced electrocatalytic oxygen evolution reactions.

Recently, the design and synthesis of Co9S8 micro/nanostructures have attracted attention as electrochemical energy storage and conversion devices due to their low cost and environmental friendliness. Herein, Co9S8 nanorings were synthesized via a one-step solvothermal method with the incorporation of Fe ions, subsequently, properly selenized to boost their electrocatalytic performance. The morphology and structure of the series of cation and anion regulated Co9S8 nanorings were characterized, the electrochemical oxygen evolution reaction (OER) properties were assessed. It is worth noting that the as-prepared catalysts, especially the innovative Fe and Se ions double doped Co9S8 nanorings, denoted as Se/Fe-Co9S8-0.14, exhibited good electrocatalytic OER performance with low overpotential (298 mV) and high durability under alkaline conditions. This work provides a new perspective to develop non-noble metal Co9S8-based OER electrocatalysts with a superior electrocatalytic performance.

Exploration of Plastid Phylogenomic Conflict Yields New Insights into the Deep Relationships of Leguminosae.

Phylogenomic analyses have helped resolve many recalcitrant relationships in the angiosperm tree of life, yet phylogenetic resolution of the backbone of the Leguminosae, one of the largest and most economically and ecologically important families, remains poor due to generally limited molecular data and incomplete taxon sampling of previous studies. Here, we resolve many of the Leguminosae's thorniest nodes through comprehensive analysis of plastome-scale data using multiple modified coding and noncoding data sets of 187 species representing almost all major clades of the family. Additionally, we thoroughly characterize conflicting phylogenomic signal across the plastome in light of the family's complex history of plastome evolution. Most analyses produced largely congruent topologies with strong statistical support and provided strong support for resolution of some long-controversial deep relationships among the early diverging lineages of the subfamilies Caesalpinioideae and Papilionoideae. The robust phylogenetic backbone reconstructed in this study establishes a framework for future studies on legume classification, evolution, and diversification. However, conflicting phylogenetic signal was detected and quantified at several key nodes that prevent the confident resolution of these nodes using plastome data alone. [Leguminosae; maximum likelihood; phylogenetic conflict; plastome; recalcitrant relationships; stochasticity; systematic error.].

Screening of Concentration and Antimicrobial Effectiveness of Antimicrobial Preservative in Betastatin Besylate Nasal Spray.

OBJECTIVE: To explore the optimal concentration and antimicrobial effectiveness of antimicrobial preservative in betastatin besylate nasal spray. METHODS: By using Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, and Aspergillus niger as test strains, the antimicrobial effectiveness of betastatin besylate nasal spray containing different concentrations of antimicrobial preservative (0.02%, 0.0125%, and 0.005% benzalkonium chloride, respectively) was determined by using bacteriostatic effect test (Chinese Pharmacopoeia, 2015 edition). RESULTS: The antimicrobial effectiveness of betastatin besylate nasal spray containing 0.02% and 0.0125% benzalkonium chloride, respectively, complied with the regulations of Chinese Pharmacopoeia (2015 Edition) against five test strains. However, the antimicrobial effectiveness of betastatin besylate nasal spray containing 0.005% benzalkonium chloride against P. aeruginosa did not meet the requirements of Chinese Pharmacopoeia. CONCLUSION: Benzalkonium chloride at a concentration of 0.125% can be used as an added antimicrobial preservative in betastatin besylate nasal spray.

Plastid Genome Evolution in the Early-Diverging Legume Subfamily Cercidoideae (Fabaceae).

The subfamily Cercidoideae is an early-branching legume lineage, which consists of 13 genera distributed in the tropical and warm temperate Northern Hemisphere. A previous study detected two plastid genomic variations in this subfamily, but the limited taxon sampling left the overall plastid genome (plastome) diversification across the subfamily unaddressed, and phylogenetic relationships within this clade remained unresolved. Here, we assembled eight plastomes from seven Cercidoideae genera and conducted phylogenomic-comparative analyses in a broad evolutionary framework across legumes. The plastomes of Cercidoideae all exhibited a typical quadripartite structure with a conserved gene content typical of most angiosperm plastomes. Plastome size ranged from 151,705 to 165,416 bp, mainly due to the expansion and contraction of inverted repeat (IR) regions. The order of genes varied due to the occurrence of several inversions. In Tylosema species, a plastome with a 29-bp IR-mediated inversion was found to coexist with a canonical-type plastome, and the abundance of the two arrangements of isomeric molecules differed between individuals. Complete plastome data were much more efficient at resolving intergeneric relationships of Cercidoideae than the previously used selection of only a few plastid or nuclear loci. In sum, our study revealed novel insights into the structural diversification of plastomes in an early-branching legume lineage, and, thus, into the evolutionary trajectories of legume plastomes in general.

Interleukin-1ß induces the upregulation of caveolin-1 expression in a rat brain tumor model.

The aim of the present study was to investigate the expression of caveolin-1 in rat brain glioma tissue, and to determine whether interleukin-1ß (IL-1ß) has a role in this process. Using glioma cells, a tumor-burdened rat model was established, and the expression of caveolin-1 protein in the tumor sites was significantly increased following intracarotid infusion of IL-1ß (3.7 ng/kg/min), as indicated by western blot analysis. The maximum value of the caveolin-1 expression was observed in tumor-burdened rats after 60 min of IL-1ß perfusion, and which was significantly enhanced by vascular endothelial growth factor (VEGF). In addition, VEGF also significantly increased IL-1ß-induced blood tumor barrier (BTB) permeability. The results suggest that the IL-1ß-induced BTB permeability increase may be associated with the expression of caveolin-1 protein, and VEGF may be involved in this process.

Cyclooxygenase inhibitor induces the upregulation of connexin-43 expression in C6 glioma cells.

The present study was performed to determine whether aspirin, a cyclooxygenase (COX) inhibitor, has an effect on the expression of connexin 43 (Cx43) in C6 glioma cells. Using an in vitro glioma invasion model, the expression of Cx43 protein in C6 cells was significantly increased following aspirin treatment at a dose of 8 mmol/l for 30, 60 and 120 min via western blot analysis. The peak value of the Cx43 expression was observed in C6 cells after 120 min of aspirin treatment, which was significantly reduced by prostaglandin E2 (PGE2). In addition, aspirin also significantly increased the gap junction intercellular communication (GJIC) activity and reduced glioma invasion, which was induced by PGE2. This led to the conclusion that the aspirin-induced glioma invasion decrease may be associated with the increased expression of Cx43 protein and formation of GJIC.

miR-381 suppresses C/EBPα-dependent Cx43 expression in breast cancer cells.

Cx43 (connexin43) is an enhancer of the metastasis of breast cancer cells. Our previous study identified miR-381 as an indirect suppressor of Cx43 gene expression, with the precise mechanism being not understood. In the present study, using a reporter gene assay, we found that miR-381 suppressed Cx43 gene expression via the promoter region -500/-250. With site-directed gene mutation, we demonstrated that miR-381 could directly bind with the sequences CACUUGUAU in the 3'-UTR so as to inhibit C/EBPα (CCAAT/enhancer-binding protein α) expression. C/EBPα was further identified as a novel transcription factor by binding to a canonic element (AATTGTC) locating at -459/-453 in the promoter region of the Cx43 gene. Functionally, we demonstrated that miR-381 suppressed C/EBPα- and Cx43-dependent migration and invasion of breast cancer cells. Finally, we revealed that decreased levels of miR-381 as well as increased expression of C/EBPα and Cx43 in the metastatic breast cancer cells and tissues. Therefore we are the first to identify that miR-381 suppresses C/EBPα-dependent Cx43 expression in breast cancer cells. The miR-381-C/EBPα-Cx43 axis might be a useful diagnostic and therapeutic target of metastatic breast cancer.

Identification of miR-200a as a novel suppressor of connexin 43 in breast cancer cells.

Both miRNAs (miRs) and connexin 43 (Cx43) were important regulators of the metastasis of breast cancer, whereas the miRs regulating Cx43 expression in breast cancer cells were still obscure. In the present study, we scanned and found miR-1, miR-206, miR-200a, miR-381, miR-23a/b and miR-186 were functional suppressors of human Cx43 mRNA and protein expression. Specially, we demonstrated that only miR-200a could directly target the 3'-untranslated region (3'-UTR) of human Cx43 gene. Functionally, overexpression of Cx43 in MCF cells potentiated the migration activity, whereas additional miR-200a treatment notably prevented this effect. Finally, we demonstrated that decreased levels of miR-200a and elevated expression of Cx43 in the metastatic breast cancer tissues compared with the primary ones. Thus, we are the first to identify miR-200a as a novel and direct suppressor of human Cx43, indicating that miR200a/Cx43 axis might be a useful diagnostic and therapeutic target of metastatic breast cancer.

[Morphological observation of human gastric cancer cell SGC-7901 clones and identification of gastric cancer stem cells].

OBJECTIVE: To dynamically investigate the morphology of human gastric cancer SGC-7901 cell clones, and then compare the tumorigenic ability of different clones in order to identify the tumor stem cell clones. METHODS: Clones derived from gastric cancer SGC-7901 cells were assessed by morphological observation, and the clone formation rate and proportion of each clone were calculated. The expression of CD44 and CDX2 in different clones was detected by immunofluorescence microscopy and Western blot. Furthermore, different clones were isolated and cultured, and their self-renewal property was assayed. Cells of different clones were subcutaneously inoculated into nude mice and the tumorigenic ability of each group was determined. RESULTS: Clones derived from gastric cancer SGC-7901 cells had three types, i.e. clones of tight, transitional and loose types. The total clone formation rate was (9.80 ± 1.07)%, and the proportion of tight, transitional and loose type clones was 10.2%, 56.0% and 33.8%, respectively. The results of immunofluorescence microscopic examination showed that the signal of CD44 was significantly stronger in the tight clones than in the transitional and loose clones, however, the signal of CDX2 was weakest in the tight colonies. The results of Western blot were consistent with that of immunofluorescence microscopic observation. SGC-7901 cells of tight clones possessed strong ability of self-renewal and in vivo tumorigenicity in the nude mice. CONCLUSION: SGC-7901 cell clones vary in morphology and differentiation, and the tight type clones may include rich gastric cancer stem cells.

[Prokaryotic expression of Leptospira interrogans groEL gene and immunoprotection of its products in hamsters].

OBJECTIVE: To construct a prokaryotic expression system of groEL gene of Leptospira interrogans serogroup Icterohaemorrhagia serovar Lai strain Lai, and to determine the immunoprotective effect of recombinant GroEL protein (rGroEL) in LVG hamsters. METHODS: The groEL gene was amplified by high fidelity PCR and the amplification products were then sequenced. A prokaryotic expression system of groEL gene was constructed using routine genetic engineering technique. SDS-PAGE plus Bio-Rad Gel Image Analyzer was applied to examine the expression and dissolubility of rGroEL protein while Ni-NTA affinity chromatography was used to extract the expressed rGroEL. The immunoprotective rate in rGroEL-immunized LVG hamsters was determined after challenge with L.interrogans strain Lai. The cross agglutination titers of sera from immunized hamsters with different L.interrogans serogroups were detected using MAT. RESULTS: The nucleotide and amino acid sequences of the cloned groEL gene were the same as those reported in GenBank. The constructed prokaryotic expression system of groEL gene expressed soluble rGroEL. The immunoprotective rates of 100 and 200 µg rGroEL in LVG hamsters were 50.0 % and 75.0%, respectively. The sera from the rGroEL-immunized LVG hamsters agglutinated all the L.interrogans serogroups tested with different levels. CONCLUSION: The GroEL protein is a genus-specific immunoprotective antigen of L.interrogans and can be used to develop an universal genetically engineering vaccine of Leptospira.

The headgroup evolution of cationic lipids for gene delivery.

Cationic lipids are one of the most widely used nonviral vectors for gene delivery and are especially attractive because they can be easily synthesized and extensively characterized. Additionally, they can best facilitate the elucidation of structure-activity relationships by modifying each of their constituent domains. The polar hydrophilic headgroups enable the condensation of nucleic acids by electrostatic interactions with the negatively charged phosphate groups of the genes, and further govern transfection efficiency. The headgroups of cationic lipids play a crucial role for gene delivery; they can be quaternary ammoniums, amines, aminoacids or peptides, guanidiniums, heterocyclic headgroups, and some unusual headgroups. This review summarizes recent research results concerning the nature (such as the structure and shape of cationic headgroup) and density (such as the number and the spacing of cationic headgroup) of head functional groups for improving the design of efficient cationic lipids to overcome the critical barriers of in vitro and in vivo transfection.

Role of angiotensin-converting enzyme (ACE and ACE2) imbalance on tourniquet-induced remote kidney injury in a mouse hindlimb ischemia-reperfusion model.

In this study, the relationship between the local imbalance of angiotensin converting enzymes ACE and ACE2 as well as Ang II and Ang (1-7) and renal injury was observed in the different genotypes mice subjected to tourniquet-induced ischemia-reperfusion on hind limbs. In wild-type mice, renal ACE expression increased while renal ACE2 expression decreased significantly after reperfusion, accompanied by elevated serum angiotensin II (Ang II) level and lowered serum angiotensin (1-7) (Ang (1-7)) level. However, renal Ang (1-7) also increased markedly while renal Ang II was elevated. Renal injury became evident after limb reperfusion, with increased malondialdehyde (MDA), decreased super-oxide dismutase (SOD) activity and increased serum blood urea nitrogen (BUN) and creatinine (Cr), compared to control mice. These mice also developed severe renal pathology including infiltration of inflammatory cells in the renal interstitium and degeneration of tubule epithelial cells. In ACE2 knock-out mice with ACE up-regulation, tourniquet-induced renal injury was significantly aggravated as shown by increased levels of MDA, BUN and Cr, decreased SOD activity, more severe renal pathology, and decreased survival rate, compared with tourniquet-treated wild-type mice. Conversely, ACE2 transgenic mice with normal ACE expression were more resistant to tourniquet challenge as evidenced by decreased levels of MDA, BUN and Cr, increased SOD activity, attenuated renal pathological changes and increased survival rate. Our results suggest that the deregulation of ACE and ACE2 plays an important role in tourniquet-induced renal injury and that ACE2 up-regulation to restore the proper ACE/ACE2 balance is a potential therapeutic strategy for kidney injury.