Discovery of Novel PD-L1 Small-Molecular Inhibitors with Potent In Vivo Anti-tumor Immune Activity.

Programmed death-ligand 1 (PD-L1) has surfaced as a promising therapeutic target for various cancers due to its pivotal role in facilitating tumor immune evasion. Herein, we report a series of novel small-molecule PD-L1 inhibitors exhibiting remarkable inhibitory activity against the PD-1/PD-L1 interaction (X18: IC50 = 1.3 nM) and reinstating the suppressive effect of PD-L1 on T cells (X18: EC50 = 152.8 nM). Crystallographic studies revealed the binding mode of X18 and PD-L1. Through a rational prodrug design approach, we have successfully optimized the oral pharmacokinetic properties of X22, effectively addressing the poor oral pharmacokinetic profile of PD-L1 small-molecule inhibitors. Notably, X22 demonstrated significant antitumor efficacy in murine models of MC38 and CT26 colon cancer through the upregulation of tumor infiltration and cytotoxicity of CD8+ T cells partially. These findings offer promising prospects for the advancement of PD-L1 inhibitors as innovative agents in cancer immunotherapy.

First report of Black Spot on Eucalyptus grandis × Eucalyptus urophylla caused by Pseudoplagiostoma eucalypti in Guangxi, China.

Eucalyptus grandis × Eucalyptus urophylla hybrid clone is an economically and ecologically important forest variety and is widely planted in Guangxi, China. Black spot, a newly found disease, occurred nearly 5333.3 hectares in an E. grandis × E. urophylla plantation of Qinlian forest farm (N: 21.866°, E: 108.921°) in Guangxi in October, 2019. Infected plants had lesions of black spots with watery margins on petioles and veins of E. grandis × E. urophylla. The size of spots ranged between 3 to 5 mm in diameter. When lesions expanded to girdle the petioles, wilt and death of leaves was observed, which subsequently affected growth of the trees. To isolate the causal agent, symptomatic plant tissues (leaves and petioles) were collected from two different sites, sampled from five plants each site. In the lab, infected tissues were surface sterilized with 75% ethanol for 10 seconds, then 2% sodium hypochlorite for 120 seconds, and rinsed with sterile distilled water three times. Small segments (5×5 mm) were cut from the margins of the lesions, then placed on potato dextrose agar (PDA) plates. The plates were incubated at 26°C in dark for 7 to 10 days. Fungal isolates YJ1 and YM6 with a similar morphology, which were obtained from 14 of 60 petioles and 19 of 60 veins respectively. These two colonies were initially light orange, then turned to olive brown as time progressed. Conidia were hyaline, smooth, aseptate, ellipsoidal, apex obtuse, and base tapering to flat protruding scar, 16.8 to 26.5µm long, and 6.6 to 10.4 µm wide (n=50). Some conidia had one or two guttules. The morphological characteristics were consistent with the description of Pseudoplagiostoma eucalypti Cheew., M. J. Wingf. & Crous (Cheewangkoon et al. 2010). For molecular identification, the internal transcribed spacer (ITS), ß-tubulin (TUB2) genes were amplified using primers ITS1/ITS4 and T1/Bt2b, respectively (White et al. 1990; O'Donnell et al.1998; Glass and Donaldson 1995). Sequences of the two strains were deposited in GenBank (ITS: MT801070 and MT801071; BT2: MT829072 and MT829073). Phylogenetic tree was constructed with a maximum likelihood method, revealing that YJ1 and YM6 were on the same branch with P. eucalypti. Pathogenicity tests of the two strains were performed on three-month-old E. grandis × E. urophylla seedlings, by inoculating 6 wounded (by stabbing on petioles or veins) leaves of seedlings with mycelial PDA plugs (5 ×5 mm) from the edge of a 10-day old colony of strain YJ1 or YM6. Another 6 leaves were treated in the same manner but with PDA plugs as controls. All treatments were incubated in humidity chambers at 27°C and 80% relative humidity, under ambient light. All experiments were conducted three times. Lesions were observed at the points of inoculation, the petioles or veins turned black on inoculated leaves after 7 days, wilting of the leaves were also observed after 30 days, however the controls remained asymptomatic. Re-isolation was made and the fungus had same morphological measurements as the inoculated fungus, thus completing Koch's postulates. P. eucalypti had been reported as a pathogen of leaf spot on E. robusta in Taiwan island (Wang et al. 2016), leaf and shoot blight on E. pulverulenta in Japan (Inuma et al. 2015). To our knowledge, this is the first report of P. eucalypti affecting E. grandis × E. urophylla in mainland China. This report provides basis for the rational prevention and control of this new disease in the cultivation process of E. grandis × E. urophylla.

First report of anthracnose caused by Colletotrichum siamense on Illicium verum in China.

Aniseed (Illicium verum) is a woody spice tree that has been grown in China for a long time. Anthracnose is an important disease of aniseed, which can cause severe leaf drop. In Sep. 2020, severe anthracnose was observed in Shanglin (23°35'5"N, 108°19'51"E), Nanning, Guangxi in China, and the incidence was 85%. The symptoms at the early stage were small, round and watery, then became larger and gradually turned brown. The acervuli would appear at the later stage, and contain many conidia. Leaves with disease were randomly sampled from 10 plants, and were cut into small rectangular pieces of 0.5×1 cm, and disinfected with 75% alcohol 1 min, with 0.1% HgCl2 3 min. After washing with sterile water 3 times, they were placed onto potato dextrose agar (PDA) medium and incubated at 25°C for 5 days. The average colony growth rate was 11.85 mm/d in 7 days. The colony was white or light gray in the initial stage, with dense aerial mycelium, and the central mycelium of the colony was dark grey in the later stage. Conidia were colorless, single spore, smooth, cylindrical, both ends obtuse, with an average size of 14.95 ± 0.97 µm × 5.46 ± 0.44 µm (n = 100). The conidial appressorium was oval or club-shaped, brown, margin intact, with an average size of 7.83 ± 1.21 µm × 5.82 ± 0.58µm (n = 50). Three strains GXNN02, GXNN03 and GXNN05 were selected for further study. Primer pairs T1/ßt2b, ACT512/ACT783, GDF/GDR, CHS1-79F / CHS1-354R and ITS1/ITS4 (Weir et al. 2012) were used to amplify tubulin (TUB), actin (ACT), 3-phosphate glyceraldehyde dehydrogenase (GAPDH), chitinase (CHS1) and the internal transcribed spacers of rDNA (ITS) respectively. BLASTn searches showed that the TUB (ON619861-63 ), ACT (ON619852-54), GAPDH (ON619855-57), CHS1 (ON619858-60) and ITS (ON573028-30) sequences had the highest similarity to Colletotrichum siamense with up to 99% (699/702, 676/679, 699/702) identity for TUB (JX010404.1); 99% (281/282, 253/254, 249/250) identity for ACT (JX009518.1); 99% (275/277, 275/277, 239/241) identity for GAPDH (JX009924.1); 99% (296/299, 296/299, 259/262) identity for CHS1 (JX009865.1); up to 99% (527/530, 485/487, 527/530) identity for ITS (JX010171.1) of ex-type ICMP 18578. A ML tree was constructed by combining 5 sequenced loci, and three isolates clustered in the C. siamense clade with 94% bootstrap support. Therefore, combined with the morphological characteristics, the pathogens were identified as C. siamense. In a pathogenicity test, these three isolates were tested on 9 healthy aniseed seedlings with at least 10 leaves, and 3 seedlings as control. The leaves were surface disinfected with 75% alcohol, and then wiped with sterilized water three times. Holes were made near the edge of the leaves and were sprayed with conidial solution (6×106 spores/mL) in test groups, and use sterilized water as control. Then the leaves were sealed inside a plastic bag for 48 h to retain moisture. Brown spot and black acervuli, similar to the symptoms in the field, were observed on the leaves in test groups within 10-15 days. No symptoms were observed on the negative control leaves. The pathogens were reisolated from the treated infected leaves and were identified as C. siamense, thus fulfilling Koch's postulates. The pathogenicity test was confirmed by repeating in triplicate. The isolation frequency of C. siamense in our samples was 82.50%. To our knowledge, this is the first report of C. siamense in China. Further research on the occurrence of the disease will help prevent the spread of the disease.

Closure of Complex Wounds by a Simple Skin Stretching System Associated With Vacuum Sealing Drainage-Clinical Outcome of 34 Patients.

Management of complex wounds with large skin defects presents a real challenge for orthopedic or reconstructive surgeons. We developed a simple skin stretching system associated with vacuum sealing drainage to examine the efficiency and complication. A total of 34 patients with different types of complex wounds were retrospectively included from January 2015 to March 2021. All patients in the study were underwent the treatment by 2 stages. The method was used to the wounds from 4.71 to 169.65 cm2 with a median defect size of 25.13 cm2. The median time for wound closure was 11.5 days (range: 5-32 days), although the median absolute reduction was 2.08 cm2/day (range: 0.15-25.66 cm2/day). Depending on the site of the wounds, the cause of the wound, and the rate of max-width/max-length (W/L), these complex wounds could be separately divided into several groups. There were statistically significant differences in the median value of the above variables (P < .05 Kruskal-Wallis test). The results showed that different anatomical sites had different viscoelastic properties, the complex wounds caused by trauma were easier to close than caused by diabetic foot and the complex wounds in group A (W/L ＞ 0.5) were more difficult to close than in group B (W/L ≤ 0.5). No major complications were encountered in this study. In summary, the results of our study showed that the simple skin stretching system associated with vacuum sealing drainage was a safe approach for closure of complex wounds. Nevertheless, more attention should be paid to the viscoelasticity of the wounds to ensure closure and avoid undue complications when applying the method.

First report of leaf blight on Eucalyptus cloeziana caused by Coniella quercicola in China.

Eucalypt species are among the most important for timber production worldwide. Eucalyptus cloeziana is increasingly culticated due to its desirable structural properties. Leaf blight is one of the most devastating diseases of E. cloeziana in China. In May 2019, leaf blight samples were collected from E. cloeziana in Chongzuo, Guangxi, China (22°20'37.70"N, 107°49'29.29"E). Lesions began at the leaf margin and extended to 1/4-3/4 of the total leaf surface area. Lesions (26.76±12.64 mm diameter) were round, yellow, and withered in appearance, and sometimes many black, round pycnidia were observed. Leaves with blight were collected randomly from 10 E. cloeziana plants. Tissue blocks (3 mm×3 mm) were sampled from diseased and healthy leaf portions, then surface disinfected with 75% ethanol for 20 s and 0.1% HgCl2 for 3 min. After washing with sterile water three times, dry tissue blocks were placed on potato dextrose agar (PDA) medium and incubated at 28°C for 5 days. Hyphae were milky white or whitish, and sparse. The colonies had petal-shaped edges and the conidiophores were clustered, branched and transparent. Spore-forming cells were solitary and smooth; conidia were smooth, fusiform or oblong, transparent, blunt-based, mostly erect, and 16.54±2.19 × 3.38±0.77 µm (n=100 in each isolate) in size. Three representative isolates (AB-6, AB-9, AB-16) were selected for further study. For molecular identification, the internal transcribed spacer (ITS) region of rDNA, translation elongation factor 1-α (TEF1), and large subunit ribosomal RNA (LSU) were amplified with primers ITS1/ITS4 (White et al. 1990), EF1-983F/EF1-1567R (Rehner and Buckleyet al. 2005), and LR0R/LR5 (Vilgalys and Hesteret al. 1990), respectively. BLASTn searches showed that the ITS (OM280456, ON026088-89), TEF1 (ON055278-80) and LSU (OM281346, ON026097-98) sequences had the highest similarity to Coniella quercicola strains with: 99% (600/605, 600/605, 600/604) identity for ITS (MH859478.1); 98% (326/333, 327/334, 325/332) identity for TEF1 (KX833698.1); 99% (870/872, 833/834, 830/831) identity for LSU (MH871258.1) of ex-type CBS 904.69. A Neighbor-Joining phylogenetic tree was constructed by combining 3 sequenced loci. Three isolates clustered in the C. quercicola clade with 100% bootstrap support. Thus, based on morphological (Maas et al. 1979; Wang and Lin et al. 2004) and molecular characteristics, the pathogen was identified as C. quercicola. In a pathogenicity test, 20 healthy E. cloeziana seedlings with at least 5 leaves were divided into 4 groups: groups 1-3 were used to inoculate three isolates respectively, and the fourth group acted as control. After surface disinfection with 75% ethanol and wiping with sterile water, tiny wounds were maked made by inoculation needle on each leaf. Fungal culture plugsblocks cut from 3 isolates were placed on wounds in groups 1-3 respectively,. withWarter- agar blockplugs served as control in group 4. The leaves were covered with wet cotton and sealed in airtight bags to retain moisture at room temperature with natural light. After 3 days, light brown lesions were observed in groups 1-3, with no symptoms present in the control group. The pathogenicity test was confirmed by repeating in triplicate and fungi re-isolated from symptomatic leaves were identified as C. quercicola. To our knowledge, this is the first report of leaf blight on E. cloeziana caused by C. quercicola in China. This study increases our understanding of E. cloeziana leaf blight and future research may allow the development of targeted prevention methods for more effective disease controls.

First report of branch blight caused by Diaporthe ueckerae on Eucalyptus citriodora in China.

Eucalyptus citriodora is a wood and oil dual-purpose tree with strong growth adaptability and high ornamental value. Recent years, it has been widely planted in Guangxi in China. In Nov. 2021, branch blight was found to be widespread on E. citriodora in Qinzhou in China (21°57'57"N, 108°42'6"E). The occurrence area was over 7000 m2, and the disease incidence was 23% (23/100). Twigs were withered in most of infected plants, and only top branch died in few plants. The lesions started from branch tips, then expanded and caused 5-20 cm branches died in final. The lesions were tiny and brown at early stage, then turned dark brown or black. Ten diseased branches were sampled randomly in field and were cut into 1 cm pieces. After surface disinfecting with 75% ethanol for 3 min, 0.1% HgCl2 for 5 min, washing with sterile water three times, samples were placed onto Potato Dextrose Agar (PDA) medium. Hyphae appeared after incubating for 5 days at 28 ℃. The diameter of colonies reached 64-71 mm after 7 days incubation. The colonies were white and felt-like, and then turned yellowish gradually with flourish aerial hyphae. Two weeks later, pycnidia appeared, which were nearly spherical, initial pale yellow and later black. Sometimes secretions overflowed from the aperture on pycnidia. There were 2 types of conidia (α and ß type), which were unicellular and hyaline. The α type was spindle to ellipse and had 1-2 oil globule, with 5.56±0.50 × 2.67±0.39 µm (n=100) in size. The ß type was linear and one end bent in hook shape, with 17.10±2.54 × 1.55±0.32 µm (n=50) in size. Three isolates (LEQZ01, LEQZ02, LEQZ03) were selected for further study. The internal transcribed spacers (ITS) region of rDNA, translation elongation factor 1-α (tef1 α) and ß-tubulin (tub2) genes were amplified using primer pairs ITS1/ITS4 (White et al. 1990), EF1-983F/EF1-1567R (Rehner et al. 2005) and Tub1/ Tub2 (Chauhan et al. 2007), respectively. BLASTn searches showed that the ITS (OM339849, ON075781, ON075782), tef1 α (ON093807-ON093809) and tub2 (ON093810-ON093812) sequences had the highest similarity with Diaporthe ueckerae strains, with 99% (543/549, 544/549, 544/549) identity for ITS (NR 147543.1), with 99% (350/351, 353/353, 349/350) identity for tef1 α (KY569388.1), with 99% (754/754, 753/754, 754/754) identity for tub2 (MW514128.1). A neighbor-joining tree constructed by combining 3 sequenced loci. Three isolates clustered in the D. ueckerae clade with 100% bootstrap support. Based on morphological (Yi et al. 2018) and molecular evidences, the pathogen was identified as D. ueckerae. In a pathogenicity test, 20 healthy E. citriodora seedlings were divided into 4 groups. Before inoculation, twigs were surface disinfected with 75% ethanol followed by washing 3 times using sterile water. Tiny artificial wounds at 5 cm below the seedling top were inoculated by hyphae taken from colonies incubated for 7 days at 25 ℃ in the dark, and covered with damp cotton in 1-3 groups (Yi et al. 2018). Yet the wounds were covered with damp cotton in control group. Two days later, wounds started to turn brown in test groups, and symptoms similar to field were obtained after 10 days. But no lesion emerged in control group. Then germs were re-isolated from symptomatic twigs and identified as D. ueckerae following the methods above. To our knowledge, this is the first report of branch blight caused by D. ueckerae on E. citriodora in China. Further researches on disease epidemiology would help to prevent spread to more locations.

First report of leaf blight caused by Apoharknessia eucalyptorum on Eucalyptus grandis × Eucalyptus urophylla in China.

Eucalypt GL-9 (Eucalyptus grandis × Eucalyptus urophylla) is one of the most widely grown genotypes of Eucalyptus in China. Each year, leaf blight causes serious economic losses in the eucalyptus industry in the south of China. In December 2019, a leaf blight disease was found to be widespread on eucalyptus GL-9 in Hechi in Guangxi, China (25°22'17"N, 108°15'32"E). Symptomatic lesions were usually brown at the early stage of infection and then turned off-white at the late stage. They had a large number of black round pycnidia randomly dispersed on the surface. Most of the lesions initially started from the leaf tip and then gradually expanded to the base of the leaf. Three randomly sampled leaves were washed using sterile water. Next, small pieces of tissue (5×10 mm) were removed from the margins of the lesions, surface disinfected with 75% ethanol for 1 min and 0.1% HgCl2 for 3 min, and then washed three times with sterile water. The tissues were placed on potato dextrose agar (PDA) and incubated at 28°C for 5 days to observe the fungus morphological characteristics. The hyphae on the PDA were milky yellow, and the PDA was light yellow when viewed from the bottom, with few aerial hyphae. The colonies had petal-like edges. In the later stage, hyphae in the center of the colonies turned brown. Three representative isolate (EC7, EC8, EC10) were selected for further study. Their conidia were olive-shaped, spindle-shaped, or obliquely globose, 8.80-11.93 µm in length (10.35 µm in average), 4.69-7.33 µm in width (6.06 µm in average) (n=100 in each isolate), with a conical apiculus and a hyaline basal appendage that was tubular, smooth, and thin-walled. For molecular identification, their genomic DNA was extracted using a Genomic DNA Kit (Tiangen, China). The internal transcribed spacer (ITS) region of rDNA and ß-tubulin (TUB) genes were amplified using ITS5/ITS4 and ßt2a/ßt2b primer sets, respectively (White et al. 1990). BLASTn searches showed that the ITS and TUB sequences had the highest identity with Apoharknessia eucalyptorum strains, with 100% (586/586 in EC7 and EC8, 590/590 in EC10) identity for ITS (KY979752.1) and 99% (502/505 in EC7, 506/508 in EC8 and 504/507 in EC10) identity for TUB (KY979919.1) of ex-type CBS 142519. The ITS and TUB sequences of three isolates were submitted to GenBank (EC7: OM060439 and OM103586, EC8: OM679378 and OM715153, EC10: OM679377 and OM715152). A maximum likelihood phylogenetic tree was constructed by combining the two sequenced loci in MEGA7. Three isolates clustered in the A. eucalyptorum clade with 92% bootstrap support. Thus, based on morphological (Crous et al. 2017; Garrett et al. 2018) and molecular characteristics, the pathogen was identified as A. eucalyptorum. In a pathogenicity test, twenty healthy GL-9 seedlings were collected, and were divided into four groups. Seedlings from groups 1-3 were used to inoculate three isolates respectively, and seedlings from another group were sprayed distilled water as control. Before test, leaves were washed with sterile water, surface disinfected with 75% ethanol, and then rinsed with sterile water. After drying, an inoculation needle was used to make tiny wounds near the leaf margin on each leaf. Next, conidia solution (1×107 conidia/ml) and sterile water were sprayed to leaves in different groups and moistened with airtight bags. After 3 days, airtight bags were moved. Lesions appeared on all the pathogen-inoculated leaves, whereas only the inoculation point turned brown on the control leaves. The pathogenicity test was repeated three times and the same results were obtained. Fungi were re-isolated from symptomatic leaves and identified as A. eucalyptorum following the same methodologies used for the initial identification. To our knowledge, this is the first report of A. eucalyptorum causing leaf blight on E. grandis × E. urophylla in China. This study expands the understanding of the pathogen of leaf blight on E. grandis × E. urophylla. More research is needed to develop effective strategies to manage this disease.

Syntheses, Biological Evaluations, and Mechanistic Studies of Benzo[c][1,2,5]oxadiazole Derivatives as Potent PD-L1 Inhibitors with In Vivo Antitumor Activity.

A series of novel benzo[c][1,2,5]oxadiazole derivatives were designed, synthesized, and biologically evaluated as inhibitors of PD-L1. Among them, compound L7 exhibited 1.8 nM IC50 value in a homogeneous time-resolved fluorescence (HTRF) assay, which was 20-fold more potent than the lead compound BMS-1016. In the surface plasmon resonance (SPR) assay, L7 bound to human PD-L1 (hPD-L1) with a KD value of 3.34 nM, without showing any binding to hPD-1. In the cell-based coculture assay, L7 blocked PD-1/PD-L1 interaction with an EC50 value of 375 nM, while BMS-1016 had an EC50 value of 2075 nM. Moreover, compound L24, an ester prodrug of L7, was orally bioavailable and displayed significant antitumor effects in tumor models of syngeneic and PD-L1 humanized mice. Mechanistically, L24 exhibited significant in vivo antitumor effects probably through promoting antitumor immunity. Together, this series of benzoxadiazole PD-L1 inhibitors holds promise for tumor immunotherapy. Preclinical trials with selected compounds are ongoing in our laboratory.

Application of a simple skin stretching system and negative pressure wound therapy in repair of complex diabetic foot wounds.

The management of complex diabetic foot wounds with large skin defects poses a challenge for surgeons. We presented a simple skin stretching system and negative pressure wound therapy for the repair of complex diabetic foot wounds to examine the effectiveness and safety.A total of 16 patients with diabetic foot ulcers were retrospectively reviewed between January 2015 and October 2020. All patients underwent the treatment by 3 stages. In stage 2, these difficult-to-close wounds of diabetes foot were residual. This method was applied to the wounds with a median defect size of 20.42 cm2 (range, 4.71-66.76 cm2).The median time for closure of complex diabetic foot wounds was 14 days ranging from 8 to 19 days. With respect to the absolute rates of reduction, it was observed with a median of 1.86 cm2/day, ranging from 0.29 cm2/day to 8.35 cm2/day. In accordance with the localization of the defect, the patients were divided into 3 groups: side of the foot (37.5%), dorsum of the foot (50.0%), and others (12.5%). There was no statistical difference between side of the foot and dorsum of the foot in terms of the median defect size with P = 0.069 (Kruskal-Wallis test). Otherwise, there were statistically significant differences regarding the median time and the median absolute rates (P < 0.05; Kruskal-Wallis test). No severe complications were encountered in this study.In summary, our results show that application of the simple skin stretching system and NPWT is an effective and safe approach to complex diabetic foot wounds. Nevertheless, more attention should be paid to the appropriate patient selection and intraoperative judgment to ensure wound closure and avoid undue complications.

Fabrication of superconducting niobium nitride nanowire with high aspect ratio for X-ray photon detection.

The niobium nitride (NbN) nanowires fabricated with the high-quality ultra-thin NbN film with a thickness of 3 nm-6 nm were widely used for single photon detectors. These nanowires had a low aspect ratio, less than 1:20. However, increasing the thickness and the aspect ratio of highly-uniformed NbN nanowires without reducing the superconductivity is crucial for the device in detecting high-energy photons. In this paper, a high-quality superconducting nanowire with aspect ratio of 1:1 was fabricated with optimized process, which produced a superconducting critical current of 550 µA and a hysteresis of 36 µA at 2.2 K. With the optimization of the electron beam lithography process of AR-P6200.13 and the adjustion of the chamber pressure, the discharge power, as well as the auxiliary gas in the process of reactive ion etching (RIE), the meandered NbN nanowire structure with the minimum width of 80 nm, the duty cycle of 1:1 and the depth of 100 nm were finally obtained on the silicon nitride substrate. Simultaneously, the sidewall of nanowire was vertical and smooth, and the corresponding depth-width ratio was more than 1:1. The fabricated NbN nanowire will be applied to the detection of soft X-ray photon emitted from pulsars with a sub-10 ps time resolution.

[Biomechanical study on femoroplasty-augmentation for prevention of osteoporotic hip fractures].

OBJECTIVE: To explore whether femoral plasty can improve the fracture resistance of osteoporotic femoral specimens and prevent hip fracture, and to compare the difference of mechanical strength changes between two different femoral plasty methods in osteoporotic femoral specimens, so as to determine the best strengthening area of the plasty. METHODS: Eighteen pairs of fresh osteoporotic femur specimens were collected and divided into two groups, A and B, 9 pairs in each group. Nine fresh osteoporotic femur specimens in each group were randomly selected for enhancement, and the corresponding contralateral specimens were used as control group. In group A1, the enhancement areas were femoral head, femoral neck, femoral trochanter and subtrochantericregion. And in group B1, the enhancement areas were femoral head, femoral neck and femoral trochanter region. The amount of cement injected into the femoral neck was recorded and the surface temperature of the femoral neck was measured. All specimens were biomechanically tested under simulated falls. Load-displacement curves, final loads were recorded. The final energy and stiffness of specimens were calculated. The biomechanical differences between the specimens of the enhancement group and those of the corresponding control group were compared, and the mechanical changes of the specimens by two different enhancement methods were compared. RESULTS: Compared with the control group, the ultimate load and energy of the specimens in the enhanced group increased significantly, but the stiffness did not change significantly. There was no significant difference in final load and energy between specimens strengthened by two different methods. CONCLUSION: Femoral plasty has the advantages of minimally invasive, simple operationand remarkable effect. It can be used as a new method to prevent osteoporotic hip fracture.

LINC00324 accelerates the proliferation and migration of osteosarcoma through regulating WDR66.

Osteosarcoma (OS) is a type of malignancy featured with high morbidity and easy metastasis. Although past years have witnessed the great improvement in the treatments of OS, there remains a long way to go. Therefore, further research on the underlying molecular mechanism of OS progression is in imminent need. Long noncoding RNAs (lncRNA) are recognized as a cluster of transcripts over 200 bases. Increasing studies have unveiled their significant regulatory roles in cancers, including in osteosarcoma. Long intergenic non-protein coding RNA 324 (LINC00324) is a newly identified lncRNA exerting oncogenic functions in several cancers, but its role in OS is yet to be uncovered. Therefore, the present study planned to explore the role of LINC00324 in osteosarcoma. We first validated the upregulation of LINC00324 in OS tissues and cell lines and established its correlation with OS tumor progression and metastasis. Importantly, the prognostic significance of LINC00324 was identified in patients with OS. Gain- and loss-of-function assays revealed that LINC00324 accelerated cell proliferation and migration in OS. Mechanistically, we revealed that LINC00324 stabilized WD repeat-containing protein 66 (WDR66) messenger RNA through interacting with Hu antigen R. Rescue assays verified that WDR66 was required for the regulation of LINC00324 in promoting proliferation and migration of OS cells. In conclusion, the present study proved that LINC00324 accelerated the proliferation and migration of osteosarcoma cells through regulating WDR66, providing a new prognostic target for osteosarcoma.

Occurrence and Molecular Phylogeny of Honey Bee Viruses in Vespids.

Since the discovery that honey bee viruses play a role in colony decline, researchers have made major breakthroughs in understanding viral pathology and infection processes in honey bees. Work on virus transmission patterns and virus vectors, such as the mite Varroa destructor, has prompted intense efforts to manage honey bee health. However, little is known about the occurrence of honey bee viruses in bee predators, such as vespids. In this study, we characterized the occurrence of 11 honey bee viruses in five vespid species and one wasp from four provinces in China and two vespid species from four locations in France. The results showed that all the species from China carried certain honey bee viruses, notably Apis mellifera filamentous virus (AmFV), Deformed wing virus (DWV), and Israeli acute paralysis virus (IAPV); furthermore, in some vespid colonies, more than three different viruses were identified. In France, DWV was the most common virus; Sacbrood virus (SBV) and Black queen cell virus (BQCV) were observed in one and two samples, respectively. Phylogenetic analyses of IAPV and BQCV sequences indicated that most of the IAPV sequences belonged to a single group, while the BQCV sequences belonged to several groups. Additionally, our study is the first to detect Lake Sinai virus (LSV) in a hornet from China. Our findings can guide further research into the origin and transmission of honey bee viruses in Vespidae, a taxon of ecological, and potentially epidemiological, relevance.

The temporal resolution and single-molecule manipulation of a solid-state nanopore by pressure and voltage.

The translocation of DNA molecules through nanopores has attracted wide interest for single-molecule detection. However, the multiple roles of electric fields fundamentally constrain the deceleration and motion control of DNA translocation. In this paper, we show how a single anchored DNA molecule can be manipulated for repeated capture using a transmembrane pressure gradient. Continuously and slowly changing the magnitude of the pressure provided two opposite directions for the force field inside a nanopore, and we observed an anchored DNA molecule entering the nanopore throughout the process from tentative to total entry. The use of both voltage and pressure across a nanopore provides an alternative method to capture, detect and manipulate a DNA molecule at the single-molecule level.

MiR-149 suppresses the inflammatory response of chondrocytes in osteoarthritis by down-regulating the activation of TAK1/NF-κB.

BACKGROUND: Osteoarthritis (OA) is a degenerative joint disease that with the complication of disability, while inflammation is the important response of OA. MiR-149 was down-regulated in the OA tissues, while the potential mechanism of miR-149 in OA is unclear. METHODS: A total of 20 OA patients and 20 healthy persons were enrolled in the present study. Real-time PCR was used to detect miR-149 and the mRNA expression of TAK1, western blot was used to detect the protein expression of TAK1. Luciferase reporter assay was performed to identify the targeting relationship between miR-149 and TAK1. Elisa assay was used to identify the level of several pro-inflammatory cytokines. RESULTS: MiR-149 was down-regulated in both OA tissues and IL-1ß-induced chondrocytes, while the expression of TAK1 was opposite. TAK1 was the target gene miR-149 targets TAK1 to regulate its expression. Human normal chondrocytes subjected to IL-1ß significantly promoted the inflammatory response, and also accelerated the activation of NF-κB signaling pathway, while alternatively si-TAK1, miR-149 mimic or PDTC reversed the effects of IL-1ß. Cells transfected with miR-149 inhibitor promotes the level of inflammation cytokines, as well as the activation of NF-κB, while cells co-transfected with si-TAK1 and miR-149 inhibitor abolishes the effects of miR-149 inhibitor. CONCLUSION: MiR-149 targets TAK1 to regulate the pathogenesis of OA, among which TAK1/NF-κB signaling acted as an important pathway in the inflammatory response that induced by IL-1ß.

rhPDGF-BB combined with ADSCs in the treatment of Achilles tendinitis via miR-363/PI3 K/Akt pathway.

To investigate the mechanism of recombinant human platelet-derived growth factor-BB (rhPDGF-BB) and human adipose-derived stem cells (hADSCs) in the treatment of Achilles tendinitis. Biomechanical indices of stiffness, stress, and maximum load-to-failure were detected by biomechanical test. mRNA and protein levels of miR-363, p-PI3K/AKT, tendon-related genes Collagen I, Scleraxis (Scx), and Tenascin C (TNC) were measured by qRT-PCR and western blot. The proliferation of hADSCs was accessed by MTT assay. Biomechanical indices of stiffness, stress, and maximum load-to-failure, and mRNA and protein levels of tendon-related genes could be improved by rhPDGF-BB or hADSCs alone, and could be further improved by rhPDGF-BB + hADSCs. rhPDGF-BB and hADSCs downregulated the expression of miR-363 and upregulated the levels of p-PI3K/Akt, and rhPDGF-BB + hADSCs further strengthened these effects. In addition, rhPDGF-BB promoted the proliferation of hADSCs in vitro and upregulated the expression of tendon-related genes. miR-363 mimic downregulated the levels of p-PI3K/Akt, miR-363 inhibitor upregulated the levels of p-PI3K/Akt, and miR-363 mimic and PI3K/Akt pathway inhibitor LY294002 reversed the positive effect of rhPDGF-BB on the proliferation of hADSCs, which suggested that rhPDGF-BB promoted the proliferation of hADSCs via miR-363/PI3K/Akt pathway. Biomechanical indices and tendon-related genes could be improved by rhPDGF-BB and hADSCs. Moreover, rhPDGF-BB promoted the proliferation of hADSCs via miR-363/PI3K/Akt pathway, indicating that rhPDGF-BB combined with ADSCs could treat Achilles tendinitis via miR-363/PI3K/Akt pathway.

Comparative Transcriptomics of Buzura suppressaria (Lepidoptera: Geometridae) Assembled De Novo Yield Insights Into Response After Buzura suppressaria Nuclear Polyhedrosis Virus Infection.

Buzura suppressaria Guenee (Lepidoptera: Geometridae) is a defoliator that seriously harms eucalyptus trees in South China. Buzura suppressaria nuclear polyhedrosis virus (BsNPV) is a baculovirus that infects B. suppressaria with high specificity and efficiency. Transcriptomes of B. suppressaria were sequenced before and after BsNPV infection using an Illumina-based platform to probe for differentially expressed genes (DEGs) of B. suppressaria after viral infection. On average, ∼57.4 million high-quality clean reads were generated and assembled de novo into 69,761 unigenes. The NCBI nonredundant protein, Swiss-Prot, Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene ontology (GO), and Cluster of Orthologous Groups databases were used to annotate unigenes through NCBI BLAST; 33,575 unigenes (48.1%) were then mapped to at least one of these databases, and 4,366 unigenes (6.3%) were mapped to all databases. Differential expression analysis showed that 25,212 unigenes were upregulated and 22,880 unigenes were downregulated in at least one pairwise comparison. Control versus 48 h had more DEGs than other two pairwise comparisons in either the GO or KEGG database, because the number of regulated gene would increase as BsNPV infected more tissues and would decrease as more tissues were disabled. To ascertain B. suppressaria immune response to BsNPV infection, DEGs were annotated to the GO and KEGG databases. In total, 89 GO categories are related to immune response and 1,007 DEGs are annotated to these GO categories. Furthermore, 7 downregulated DEGs and 37 upregulated were obtained simultaneously in all three groups. These DEGs were considered to possess a central role throughout viral infection.

Moving target detection for frequency agility radar by sparse reconstruction.

Frequency agility radar, with randomly varied carrier frequency from pulse to pulse, exhibits superior performance compared to the conventional fixed carrier frequency pulse-Doppler radar against the electromagnetic interference. A novel moving target detection (MTD) method is proposed for the estimation of the target's velocity of frequency agility radar based on pulses within a coherent processing interval by using sparse reconstruction. Hardware implementation of orthogonal matching pursuit algorithm is executed on Xilinx Virtex-7 Field Programmable Gata Array (FPGA) to perform sparse optimization. Finally, a series of experiments are performed to evaluate the performance of proposed MTD method for frequency agility radar systems.

[Iincidence of postoperative delirium after hip surgery in elderly patients: a meta-analysis].

OBJECTIVE: To evaluate incidence of postoperative delirium after hip surgery in elderly patients by meta-analysis. METHODS: From January 1, 2014 to December 31, 2013, clinical literatures about postoperative delirium after hip surgery in elderly patients,were searched from the Pubmed. Literature extract table were formed according to inclusion and exclusion criteria. Stata-12.0 was applied for Meta-analysis. P was used to test heterogeneity of study, random-effect model was performed when I2 > 50%. Subgroup analysis was used according to stage of age, assessment scale of delirium and statistical area of literature. Begg test was used to test publication bias. RESULTS: Twenty-one literatures were included. Incidence of postoperative delirium after hip surgery in elderly patients by weighted and combination was 17% [95% CI (16%, 18%)]. Incidence of postoperative delirium after optional hip surgery was decreased more than emergency operation in included 5 literatures [OR = 0.32, 95% CI (0.22, 0.45)]. Incidence of postoperative delirium in patients less than 80 years old was 21% [95% CI (19%, 23%)], while 21% [95% CI (19%, 24%)] in patients more than 80 years old. Incidence of postoperative delirium in CAM evaluation scale was 23% [95% CI (21%, 26%)], while 19% [95% CI (17%, 21%)] in other evaluation scales. Incidence of postoperative delirium in Asian area was 17% [95% CI (15%, 20%)], while 23% [95% CI (21%, 25%)] in European and American area. There was no publication bias tested by Begg test (P < 0.05). CONCLUSION: Incidence of postoperative delirium after hip surgery in elderly patients increases higher, especially in emergency operation. A standardizing research method is benefit for evaluate incidence of postoperative delirium after hip surgery in elderly patients, decreasing heterogeneity and publication bias.

Apo E gene polymorphism affects development of type 2 diabetic nephropathy in Asian populations, especially in East Asians: an updated meta-analysis.

BACKGROUND: Many studies have determined the correlation between the Apolipoprotein E (APO E) gene polymorphisms and diabetic nephropathy, but their results are inconclusive. MATERIAL/METHODS: With the aim to confirm this correlation, we performed a meta-analysis of 16 studies. The dichotomous data are presented as the odds ratio (OR) with a 95% confidence interval (CI). RESULTS: The results of our study indicate that APO ε2 allele among the pooled Asian populations were more likely to show high risk of DN development (2 allele vs. ε3 allele: pooled OR =1.629, 95% CI=1.010-2.628, P=0.045). For further analysis, the APO e2 allele was associated with progress of DN in the group with duration >10 years, but not in the group with duration <10 years (ε2 allele vs. ε3 allele: pooled OR=1.920, 95% CI=1.338-2.754, P<0.001). The APO e2 polymorphism increased the susceptibility to DN in Asian population compared with healthy people (ε2 allele vs. ε3 allele: pooled OR=1.629, 95% CI=1.010-2.628, P=0.045). CONCLUSIONS: Development of DN is associated with APO E polymorphisms in Asian populations, especially in East Asians.