[Clinical management of thrombocytopenia in cirrhosis].

Thrombocytopenia is one of the common complications of cirrhotic patients, which can induce an increasing bleeding risk and closely correlate with bleeding following invasive procedures. Consequently, how to respond to thrombocytopenia is crucial for improving the prognosis of patients with cirrhosis. This article reviews the main mechanisms of cirrhosis concurrent with thrombocytopenia, as well as the corresponding clinical management strategies.

[Clinical study of 19 cases of steroid-refractory gastrointestinal acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation with fecal microbiota transplantation].

Objective: To investigate the clinical efficacy of fecal microbiota transplantation (FMT) for treating steroid-refractory gastrointestinal acute graft-versus-host disease (GI-aGVHD) . Methods: This analysis included 29 patients with hematology who developed steroid-refractory GI-aGVHD after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Huaian Hospital Affiliated to Xuzhou Medical University from March 2017 to March 2022. Among them, 19 patients underwent FMT treatment (the FMT group) and 10 patients did not (the control group). The efficacy and safety of FMT were assessed, as well as the changes in intestinal microbiota abundance, lymphocyte subpopulation ratio, peripheral blood inflammatory cytokines, and GVHD biomarkers before and after FMT treatment. Results: ① Complete remission of clinical symptoms after FMT was achieved by 13 (68.4%) patients and 2 (20.0%) controls, with a statistically significant difference (P<0.05). Intestinal microbiota diversity increased and gradually recovered to normal levels after FMT and FMT-related infections did not occur. ②The proportion of CD3(+) and CD8(+) cells in the FMT group after treatment decreased compared with the control group, and the ratio of CD4(+), regulatory T cells (Treg), and CD4(+)/CD8(+) cells increased (all P< 0.05). The interleukin (IL) -6 concentration in the FMT group was lower than that in the control group [4.15 (1.91-5.71) ng/L vs 6.82 (2.40-8.91) ng/L, P=0.040], and the IL-10 concentration in the FMT group was higher than that in the control group [12.11 (5.69-20.36) ng/L vs 7.51 (4.10-9.58) ng/L, P=0.024]. Islet-derived protein 3α (REG3α) was significantly increased in patients with GI-aGVHD, and the REG3α level in the FMT group was lower than that in the control group after treatment [30.70 (10.50-105.00) µg/L vs 74.35 (33.50-139.50) µg/L, P=0.021]. Conclusion: FMT is a safe and effective method for the treatment of steroid-refractory GI-aGVHD by restoring intestinal microbiota diversity, regulating inflammatory cytokines, and upregulating Treg cells.

[Research on the effect of PD-L1 overexpression on CLL-1 CAR-T anti-acute myeloid leukemia].

Objective: To investigate the effects of programmed death receptor ligand 1(PD-L1)on CLL-1 CAR-T against acute myeloid leukemia(AML). Methods: In this experiment, the PD-L1 expression vector was constructed, and then the lentivirus vector was packaged by three-plasmid packaging system. THP-1 monoclonal cell lines stably expressing PD-L1 were set up. The CLL-1 CAR-T was developed by our team, as the effector cell for co-culture with the THP-1 or THP1-PDL1 cell lines, respectively. Then, the LDH was tested using the kit, the supernatant cytokine was detected by CBA, and the CLL-1 CAR-T cell proliferation was demonstrated by flow cytometry(FCM)with CSFE labeled. Results: ①The PD-L1 lentivirus vector was successfully constructed, and monoclonal cell lines of THP-1 with stable PD-L1 was set up and verified by FCM and PCR. ②The overexpression of PD-L1 inhibited CLL-1 CAR-T's ability to lyse THP-1 cells(E∶F ratio 10∶1); the killing efficiency of CLL-1 CAR-T on THP1-PDL1 cells was lower than that of THP-1 cells[(15.70±9.90)% vs(51.95 ± 2.52)%, P<0.05]. ③The overexpression of PD-L1 decrease the release of cytokine[THP1-PDL1 group vs THP-1 group: IFN-γ(115.66±3.13)pg/ml vs(1708.16 ± 26.76)pg/ml, P<0.05; IL-6(17.37±0.72)pg/ml vs(124.92±4.26)pg/ml, P<0.05; IL-10(5.69±0.13)pg/ml vs(124.12±3.02)pg/ml, P<0.05]. Additionally, the proliferation of CLL-1 CAR-T was also inhibited. Conclusion: Monoclonal cell lines of THP-1 with stable PD-L1 expression were successfully constructed, and the adverse effect of PD-L1 overexpression on CLL-1 CAR-T anti-AML was confirmed, which provided a theoretical basis for the regulation of CLL-1CAR-T through the PD-1/PD-L1 pathway.

MiRNA-26a promotes angiogenesis in a rat model of cerebral infarction via PI3K/AKT and MAPK/ERK pathway.

OBJECTIVE: To investigate the specific role of microRNA-26 (miRNA-26a) in a rat model of cerebral ischemic stroke and the underlying mechanism. MATERIALS AND METHODS: A rat model of middle cerebral artery occlusion (MCAO) was established to induce permanent cerebral infarction. Neuro-behavior was observed and scored after model establishment. The expression of miRNA-26a in brain tissue and brain microvascular endothelial cells (BMECs) of rats after cerebral ischemic stroke was detected by quantitative polymerase chain reaction (qPCR). The formation of the endothelial lumen was detected by Matrigel assay after BMECs were transfected with miR-26a mimics or inhibitors. Besides, cell proliferation after miRNA-26a transfection was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The protein levels related to PI3K/AKT and MAPK/ERK signaling pathway were detected by Western blot. RESULTS: miRNA-26a expression was elevated after cerebral infarction injury. Further investigation showed that miRNA-26a mimics could promote endothelial lumen formation and cell proliferation in BMECs, while miRNA-26a inhibitor inhibited the capacity of lumen formation and cell proliferation. Notably, we found that miRNA-26a might up-regulate the expression of HIF-1a via activating the AKT and ERK1/2 pathway, thus mediating the transcriptional activity of VEGF and promoting lumen formation and cell proliferation in BMECs. CONCLUSIONS: MiRNA-26a promotes angiogenesis in a rat model of cerebral ischemic via PI3K/AKT and MAPK/ERK pathway.

LncRNA MEG3 participates in neuronal cell injury induced by subarachnoid hemorrhage via inhibiting the Pi3k/Akt pathway.

OBJECTIVE: To explore the effect of LncRNA MEG3 in the subarachnoid hemorrhage (SAH) and its underlying mechanism. PATIENTS AND METHODS: The expressions of lncRNA MEG3 in SAH patients and animal model were detected by quantitative real-time PCR (qRT-PCR). After LncRNA MEG3 was overexpressed in neurons by lentivirus, viability and apoptosis abilities were detected by cell counting kit-8 (CCK-8) assay, flow cytometry, and TUNEL assay, respectively. The apoptosis-related genes and Pi3k/Akt pathway-related proteins were further detected by a Western blot. RESULTS: The expressions of lncRNA MEG3 in SAH patients were remarkably higher than normal controls, which were positively correlated with SAH severity. After lncRNA MEG3 overexpression, neuronal cell activity was decreased and cell apoptosis was increased. Moreover, the expressions of Bax, p53, and cleaved Caspase-3 were increased, whereas the expression of Bcl-2 and Pi3k/Akt pathway-related proteins were decreased after lncRNA MEG3 overexpression. CONCLUSIONS: LncRNA MEG3 is up-regulated in SAH, which may promote SAH-induced neuronal cell injury via inhibition of the Pi3k/Akt pathway.

Study on the treatment of the p15 gene combined with Bcr-abl-specific siRNA and STI571 for chronic myeloid leukemia.

The aim of this study was to investigate the effect of the p15 gene combined with Bcr-abl-specific siRNA and STI571 on the proliferation, cell cycle and apoptosis of K562 chronic myeloid leukemia cells. Using the gene sequence results, we amplified the p15 gene from normal peripheral blood by RT-PCR, and constructed a p15-pcDNA3.1 vector. The K562 cell line with G418 resistance was screened, synthesized and transfected for bcr-abl gene fusion point for 21-nt siRNA. In p15-pcDNA3.1-K562 cells, the growth rate was slower than that of the control K562 cells, G0/G1-phase was increased and S-phase was decreased significantly. In the siRNA group, bcr-abl fusion gene expression was significantly decreased in K562 cells accompanied by the downregulation of BCL-xL protein expression and G1-phase arrest. Cell survival rate was significantly decreased compared with the sole p15-K562 cell group and the sole RNA interference-K562 cell group. In the combination of p15-pcDNA3.1-K562 cells with STI571, the proportion of apoptosis was significantly increased and the cell survival rate was significantly decreased compared with the p15-K562 cell group and STI571-K562 cell group. siRNA at 30 pM combined with 0.5 µM STI571 promoted apoptosis compared with sole application. The p15 gene combined with siRNA had a synergistic effect on the inhibition of proliferation and the induction of apoptosis in K562 cells. Exogenous p15 protein expression combined with STI571 appeared to have a synergistic effect on proliferation inhibition and apoptosis induction in K562 cells. The combination of low-dose RNA interference with STI571 showed a synergistic effect in inducing apoptosis.

A laser-Compton scattering prototype experiment at 100 MeV linac of Shanghai Institute of Applied Physics.

As a prototype of the Shanghai Laser Electron Gamma Source in the Shanghai Synchrotron Radiation Facility, an x-ray source based on laser-Compton scattering (LCS) has been installed at the terminal of the 100 MeV linac of the Shanghai Institute of Applied Physics. LCS x-rays are generated by interactions between Q-switched Nd:yttrium aluminum garnet laser pulses [with wavelength of 1064 nm and pulse width of 21 ns (full width at half maximum)] and electron bunches [with energy of 108 MeV and pulse width of 0.95 ns (rms)] at an angle of 42 degrees between laser and electron beam. In order to measure the energy spectrum of LCS x-rays, a Si(Li) detector along the electron beam line axis is positioned at 9.8 m away from a LCS chamber. After background subtraction, the LCS x-ray spectrum with the peak energy of 29.1+/-4.4|(stat)+/-2.1|(syst) keV and the peak width (rms) of 7.8+/-2.8|(stat)+/-0.4|(syst) keV is observed. Normally the 100 MeV linac operates with the electron macropulse charge of 1.0 nC/pulse, and the electron and laser collision repetition rate of 20 Hz. Therefore, the total LCS x-ray flux of (5.2+/-2.0) x 10(2) Hz can be achieved.

Development of a new reaction system for the synthesis of highly optically active alpha,gamma-substituted gamma-butyrolactones.

A highly useful method for the synthesis of optically active alpha,gamma-substituted gamma-butyrolactones has been developed. The SmI(2)-induced reductive coupling of chiral 2-alkyl acrylates derived from isosorbide with ketones in the presence of (1S)-(-)-2,10-camphorsultam as a proton source give the chiral alpha,gamma-substituted gamma-butyrolactones in good yields and high enantiomeric purities (up to >99% ee for trans and 75% ee for cis). The reaction system has been investigated with various ketones, and it is demonstrated that this system is very effective for trans-alpha,gamma-substituted gamma-butyrolactones. Both the chiral auxiliary and the hindered proton source in this system are necessary for the observed excellent ee values of the products. The absolute configuration of the trans products is assigned on the basis of the X-ray crystal structure.

A highly efficient asymmetric synthesis of optically active alpha, gamma-substituted gamma-butyrolactones using a chiral auxiliary derived from isosorbide.

Using an easily accessible and inexpensive chiral auxiliary derived from isosorbide, optically active alpha,gamma-substituted gamma-butyrolactones were obtained in high enantiomeric purity (up to >99% ee for trans) by the SmI(2)-induced reductive coupling of chiral methacrylate 7 with ketones in the presence of (-)-sultam as a proton source.

Microbial transformation of 2-hydroxy and 2-acetoxy ketones with Geotrichum sp.

Biotransformation of a series of o-, m- and p-substituted alpha-hydroxy- and alpha-acetoxyphenylethanones 1a-h and 9a-g with Geotrichum sp. led to the corresponding 1,2-diols 2 and/or monoacetates 10 in moderate to excellent enantiomeric excesses. Alpha-hydroxy- and alpha-acetoxyphenylethanones and their m- and p-derivatives gave preponderantly the S-configuration products while in the case of the o-derivatives R-alcohol was provided as the major enantiomer. The results of stereoselectivity were discussed.

An efficient and convenient approach to the total synthesis of sphingofungin.

As a new member of the sphingofungin family, sphingofungin F exhibits interesting physiological activities with a structural unit of an alpha-substituted alanine. Described herein is an efficient and convenient stereoselective synthesis of sphingofungin F from L-(+)-tartaric acid, which utilizes Sharpless asymmetric epoxidation of allylic alcohol and Lewis acid-catalyzed intramolecular epoxide-opening reaction with an N-nucleophile, to introduce the other two desired stereogenic centers. Side chain functionality was incorporated into the chiral segment using a Wittig reaction.

Enzymatic synthesis of (2E,4E)-(6R,10R)-4,6,10,12-tetramethyl-2,4-tridecadien-7-one, the sex pheromone of Matsucoccus matsumurae Japanese pine bast scale.

The sex pheromone of Matsucoccus matsumurae Japanese pine bast scale (2E,4E)-(6R,10R)-4,6,10,12-tetramethyl-2,4-tridecadien-7-one (1) was synthesized with stereocontrol from (2R,4S)-5-acetoxy-2,4-dimethyl-pentanol (3), which in turn was prepared by lipase-catalyzed transesterification of meso-2,4-dimethyl-1,5-propanediol (2).

Sex phermone components of mulberry looper,Hemerophila atrilineata butler (lepidoptera: Geometridae).

(6Z-9S, 10R)-Epoxy-octadecene (SR-1) and (3Z, 6Z-9S, 10R)-epoxy-octadecadiene (SR-2) are sex pheromone components of the mulberry looper (MBL),Hemerophila atrilineata Butler. Compounds extracted from female MBL pheromone glands were identified by coupled gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometry. In field experiments in China,SR-2,RS-2, or both combined were hardly attractive, butSR-2 in combination withSR-1 attracted significant numbers of MBL males. Synergistic behavioral activity ofSR-1 plusSR-2, but not of corresponding antipode mixtures, indicates enantiospecificity of MBL pheromone communication. Because blends of racemic and enantiospecific (SR)1 plus2 were similarly attractive, racemic1 plus2 may have potential for mass trapping or confusion of MBL males in commercial mulberry plantations.