dc rongsheng llc fishers made in china

The S protein, the NP of SARS‐CoV‐2 WIV04 strain, and the RBDs of WIV04, Beta and Delta were expressed in 293‐FT cells. The NTD was purchased from Sino Biological Inc. (Cat. No. 40591‐V49H). The binding of COVID‐HIG (COVID‐HIG‐003) to the recombinant S protein, NTD, NP, and RBDs was analyzed using BLI with an Octet‐Red 96 device (Pall ForteBio LLC., CA, USA). All steps were performed in a black 96‐well plate with a working volume of 200 µL per well at 30 °C, with shaking at 400 rpm. By using RBDs as examples, 10 µg purified RBDs labeled with biotin per well were loaded onto streptavidin biosensors (ForteBio), which were activated in binding buffer (0.1% w/v bovine Serum Albumin (BSA), 0.01% w/v Tween‐20 in phosphate‐buffered saline) for 300 s. The sensors were then dipped into the serially diluted COVID‐HIG (0.16–20 μM) for 600 s for measurement of association kinetics after incubation for 180 s in the baseline buffer (0.1% w/v BSA, 0.01% w/v Tween 20 in PBS). Then, the dissociation was measured in a kinetic buffer (0.1% w/v BSA, 0.01% w/v Tween 20 in PBS) for 600 s. Octet Data Acquisition 9.0 was used for data analysis and curve fitting using an 1:1 model. The K

X.M.Y., H.C.Y., and R.J. are employees of China National Biotec Group Company Limited; D.Y., H.L., J.Z.W., and Y.L.D. are employees of Chengdu Rongsheng Pharmaceuticals Co., Ltd; D.Y., H.L., T.J.L., Y.Z., C.Y.L., D.M.D., D.X.F., D.B.Z., Y.L.H., T.D., Y.X., and R.Z. are employees of Beijing Tiantan Biological Products Co., Ltd; C.S.L., Y.H., Y.P., R.H., Y.T.X., L.F., X.L.L., Z.J.Z., D.M. J., F.F.W., J.H.Y., K.P., D.M.X., and Y.L.H. are employees of Sinopharm Wuhan Plasma‐derived Biotherapies Co., Ltd. Sinopharm Wuhan Plasma‐derived Biotherapies Co. Ltd. filed patents on the production method of COVID‐HIG to China National Intellectual Property Administration. All other authors declare no conflict of interest. Figure 6b,c has been amended on May 16, 2022, after initial online publication.

The No 4 dock at Jiangsu Rongsheng Heavy Industries Co Ltd"s Nantong shipbuilding base on May 26, 2012. With a dimension of 139.5*580m，the dock is equipped with a 1600-T gantry crane, the world"s largest. [Photo/chinadaily.com.cn]

China Rongsheng Heavy Industries Group Holdings Ltd, the nation"s largest private shipbuilder, may seek "cooperation with one or two ship builders" in 2013 or 2014, grasping the opportunity emerging from an industry recession, according to Xu Yifei, assistant president of Jiangsu Rongsheng.

In response to this round of recession, Rongsheng has been actively upgrading technology and design. It has also put more focus on the offshore engineering sector to further diversify its business.

Rongsheng is setting up its offshore engineering company in Singapore, aiming to take advantage of Singapore"s technology and existing market to deepen its penetration in the global offshore engineering market, according to Xu.

The company entered the marine engineering sector years ago. China"s first deepwater pipe-laying crane vessel, known as Hai Yang Shi You 201, was built by Rongsheng. The vessel can lay pipes at depths of 3000 meters and lift 4000 metric tons and will operate at the South China Sea"s Liwan 3-1 gas field.

Rongsheng"s president, Chen Qiang, said in an earlier interview that he hoped orders from marine engineering will make up about 40 percent of the company"s new orders this year.

We obtained bone marrow-derived dendritic cells (BMDCs) from BM suspensions prepared from mouse femurs as described previously (16). Exosomes were purified from the supernatants of DCs infected with T. gondii by differential centrifugation as previously described (17). Briefly, we harvested supernatants of DCs infected with T. gondii from the DC-Me49-exo group, and those of control DCs from the DC-exo group. The different supernatants were centrifuged at 500 g for 10 min to remove cell debris and other small particles, and the supernatants was collected. Then, we centrifuged the supernatant at 16,500 g for 20 min, followed by filtration through a 0.22-μm filter (Millipore Sigma). Finally, supernatant solutions were ultracentrifuged at 120,000 g for 90 min, and the exosome pellet was resuspended with an appropriate amount of PBS. We measured the protein content of exosomes using a BCA protein assay kit (Thermo Fisher, USA). Exosomes were stored at −80°C for future use or directly used in co-culture experiments.

Mouse BM–derived MDSCs were obtained as previously described (18, 19). In brief, we flushed BM cells from the femurs and tibias of approximately 5-week-old BALB/c mice. Red blood cells (RBCs) were lysed with lysis buffer (Thermo Fisher). The BM cells were then cultured in RPMI-1640 supplemented with 10% FBS, 1% penicillin-streptomycin solution and stimulated with 40 ng/mL interleukin-6 (IL-6) and granulocyte-macrophage colony stimulating factor (GM-CSF) (Rocky Hill, NJ, USA) at 37°C for 4 d in a 5% CO2-humidified atmosphere. Then, we detected unique M-MDSCs and G-MDSCs using (FITC)-CD45, (APC)-CD11b, (PE)-Ly6G, and (APC-700)-Ly6C. Uniquely DCs and macrophages were detected by (FITC)-CD45, (PC5.5)-CD11c, and (APC)-CD11b, (PB450)-F4/80. All the antibodies were purchased from Becton, Dickinson (San Jose, CA, USA). After induction and maturation, MDSCs were harvested. Then, MDSCs incubated with exosomes of DC-exo, DC-Me49-exo, and PBS, respectively. After 24 h, MDSC was collected for flow cytometry and Western blot. All cells were lysed using RIPA buffer (Beyotime Institute of Biotechnology, China) with 1 nM PMSF, and total protein concentration was determined using a BCA protein quantification kit (Thermo Fisher). Next, we loaded 5 µg total proteins per lane and resolved them on 0.6–0.8% gels by SDS-PAGE. Proteins were transferred onto PVDF membranes and blocked for 1 h with 5% non-fat milk at RT, after which the membranes were washed three times with PBST. Membranes were incubated for 1 h with monoclonal antibodies of P-JAK2, JAK2, P-STAT3, STAT3 and tubulin in a buffer containing PBST and 1% milk. All primary antibodies were purchased from CST. After washing them with PBST, we incubated membranes for 1 h with goat anti-mouse IgG-HRP (Santa Cruz, CA, USA) and detected signals on a ChemiDoc Touch Imaging (Bio-Rad, USA).

BALB/c female mice (6 weeks old) were weighed, randomly divided into three groups (n = 10 each group), and injected subcutaneously with 5 × 106 CT26 cells. When the tumor was visible, treatment was carried out according to the following design: an intra-tumoral injection with 10 μg DC-exo (DC-exo group), 10 μg DC-Me49-exo (DC-Me49-exo group), and 10 μL PBS (PBS group). Two injections were performed on day 1 and 3 after tumor visualization (8). At day 19 post inoculation, the tumor progression was monitored by using an IVIS Spectrum imaging system (IVIS Spectrum, USA). Mice were sacrificed, and blood, spleens and tumors were collected for flow-cytometry analysis. Cytokines in serum were determined by using IL-6 and GM-CSF ELISA kits (Neobioscience Technology Company).

In the geopolitical conflict over the South China Sea (SCS), fishers are at the center of Chinese and Vietnamese cartographic imaginations that define the sea as either “Chinese” or “Vietnamese” and hence tied to the disputed territories of the Paracel and Spratly Islands. While their historical presence and customary fishing rights in the SCS have been much publicized in the context of this territorial dispute, the long-standing Cham seafaring trade networks and legacy are ignored by both countries. The ethnic and national categories of Cham, Việt, and Han intersect with occupational categories such as those of fisher, trader, shipbuilder, sailor, and pirate, which in the past represented shifting, relational, and situational activities by the same people. The contemporary use of such professional and national labels produces particular political effects by projecting recent closures and enclosures onto the past, in spite of the common historical, cultural, and ethnic flows that always existed in the SCS. Rather than aiming to legitimize or delegitimize Vietnam’s or China’s territorial claims to the SCS, this article argues that seafaring narratives should be liberated from abstract, anachronistic discourses of sovereignty, territoriality, and territorial anxieties that separate the interconnected histories of the Cham, Vietnamese, and Chinese.