Research trends and hotspots of recurrent pregnancy loss with thrombophilia: a bibliometric analysis.

BACKGROUND: Thrombophilia is a group of disorders that result in a blood hypercoagulable state and induce thrombosis, which was found widely existed in recurrent pregnancy loss (RPL). More and more research about thrombophilia has been conducted but the association between thrombophilia and RPL remains uncertain. Thus, it's necessary to combine relevant literature to find the research hotspots and analyze the internal link between different study points, and then predict the development trend in RPL with thrombophilia. METHODS: Relevant articles between 1970 and 2022 were obtained from the Web of Science (WoS) database. Software VOSviewer and CiteSpace were used to perform the analysis and conduct visualization of scientific productivity and emerging trends. RESULTS: Seven hundred twenty-five articles published in recent 30 years by 3205 authors from 1139 organizations and 68 countries were analyzed. 37authors, 38 countries, and 53 organizations published papers ≥5. The United States was the most productive country and Univ Amsterdam was the most productive institution. Journal thrombosis and haemostasis had the most total citations. In keyword and clusters, factor-v-Leiden, inherited thrombophilia, activated protein-c, low-dose aspirin, molecular-weigh heparin, polymorphism had high-frequency focus on its etiology, diagnostics, and therapeutics. The strongest keyword bursts showed the research hotspots changed over time. CONCLUSIONS: There could be differences in the clinical relevance of different type of thrombophilia, as well as single and multiple thrombophilic factors. Anticoagulation and immunotherapy are currently the main treatment options. More clinical trials and basic research are expected and we should attach more attention to the whole management of in-vitro fertilization in the future.

[Prediction and modification analysis of low-affinity HLA-A*0201 restricted CTL epitopes derived from HIV-1 pol antigen].

AIM: To screen the possible HLA-A*0201 restricted low-affinity CTL epitopes derived from HIV-1 pol antigen and to predict and identify the possible change of the affinity between epitope and the HLA-A*0201 molecule when the epitope is modified. METHODS: HLA-A*0201 restricted low-affinity CTL epitopes were predicted by CTL epitope prediction software based on super motif, proteasome cleavage probability, HLA affinity and so on. The candidates were modified acid substitution and analyzed by computer. T2 cells were used to determine the peptide by amino binding affinity and HLA-A*0201-peptide complex stability. RESULTS: Among the three predicted peptides by softer ware, YVSLSFPQI (pol52-60Y1), YVSQIIEQL pol(673-681Y1), YIQKETWEA(pol548-556Y1)could bind to HLA-A*0201 with high affinity, and the dissociation time of 50% HLA-A*0201 peptide complex was over to 8 h. CONCLUSION: Our results suggest that the three predicted peptides, as modification, might be HLA-A*0201 restricted CTL epitopes.

[Cloning and expression of PbTIP analogous gene from Plasmodium berghei ANKA and preparation of its polyclonal antibody].

OBJECTIVE: To clone and express a novel protein analogous to TIP (T cell immunomodulatory protein) of Plasmodium berghei ANKA and prepare its polyclonal antibody. METHODS: The PbTIP encoding nucleotide sequence was searched from the Plasmodium berghei genomic database and amplified by PCR. The gene was sub-cloned into prokaryotic expression vector pGEX4T-1 and expressed in E.coli BL21 (DE3). After induction by IPTG, the expression of PbTIP-GST fusion protein was characterized by SDS-PAGE and Western blotting. The inclusion bodies of GST-PbTIP fusion protein were injected into BALB/c mouse. Anti-sera were identified by indirect fluorescent antibody test and western blotting. RESULTS: The PbTIP-GST fusion protein was successfully expressed in the form of inclusion bodies, by controlling the temperature and concentration of IPTG. Anti-PbTIP-GST sera were acquired with high titer. The sera specifically recognized the PbTIP with a band of 60 000 in P.berghei infected erythrocyte protein. CONCLUSION: PbTIP/GST fusion protein and polyclonal antibody have been obtained.

[Analysis on the significance of pelvic hemodynamics in efficacy evaluation of TCM treatment for chronic pelvic inflammation].

OBJECTIVE: To explore the significance of pelvic hemodynamics as an index in evaluating efficacy of TCM treatment for chronic pelvic inflammation (CPI). METHODS: Sixty patients with CPI received treatment with Penyanping, a self-formulated TCM recipe, for 30 days, and the changes of pelvic hemodynamic indexes in them were measured before and after treatment within the 3 - 7 days after menstruation using color Doppler. RESULTS: Improvement of pelvic hemodynamics indexes were shown after treatment in ovarian left arteriopalmus index, bilateral resistance index, maximal speed of left arterial blood flow and score of time-velocity, as compared with those before treatment, the difference was significant respectively (P < 0.05). CONCLUSION: Pelvic hemodynamic indexes could be taken as one of the objective parameters for evaluating efficacy of TCM treatment of CPI according to principle of activating blood circulation to remove stasis, clearing heat and detoxifying.

[Expression, purification and transduction of Tat-p53 fusion protein].

AIM: To express and purify Tat-p53 fusion protein and investigate its transduction efficiency. METHODS: The gene encoding wide-type p53 was isolated using RT-PCR from A549 cell line and cloned into pTAT-HA and pET32a prokaryotic expression vectors. Recombinant plasmids were transformed into E.coli BL21(DE3)LysS, then the transformed cells were induced with IPTG. The expression and purification of the Tat-p53 and p53 were analyzed by SDS-PAGE. BALB/c mice were immunized with purified p53 protein. The serum was isolated and the antibody specific to p53 was measured by ELISA. The transduction efficiency of Tat-p53 was detected using indirect immunofluorescence assay. RESULTS: Prokaryotic expression vectors of Tat-p53 and p53 were constructed correctly. Tat-p53 fusion protein and p53 protein were successfully expressed and purified. p53 specific mouse antiserum was obtained. IFA result indicated that Tat-p53 fusion protein transduced into HepG2 cells efficiently. CONCLUSION: The obtained Tat-p53 fusion protein may be valuable for the basic research on therapy for liver carcinoma.

Quantifying anti-HBV effect of targeted ribonuclease by real-time fluorescent PCR.

AIM: To quantify the inhibition of HBV replication by targeted ribonuclease by using real-time fluorescent PCR. METHODS: Targeted ribonuclease was introduced into 2.2.15 cells by liposome-mediated transfection or HIV-TAT mediated protein transduction. Forty-eight hours after the transfection and 24 h after the transduction, supernatants of 2.2.15 cells were collected and HBV DNA in the supernatants was quantified by real-time fluorescent PCR with a commercial kit. RESULTS: HBV DNA concentrations in the supernatants of 2.2.15 cells transfected or transducted with targeted ribonuclease were 4.9+/-2.4 x 10(8) copies/L and 8.3+/-4.0 x 10(8) copies/L, respectively. Compared with controls, transfection or transduction of targeted ribonuclease reduced HBV DNA concentration in the supernatants of 2.2.15 cells by 90.4% and 90.1%, respectively (P<0.05). CONCLUSION: Targeted ribonuclease can inhibit HBV replication in 2.2.15 cells.

Effect of vector-expressed siRNA on HBV replication in hepatoblastoma cells.

AIM: To study the effect of siRNA expressed from DNA vector on HBV replication. METHODS: Human U6 promoter was amplified from genomic DNA and cloned into plasmid pUC18 to construct a mammalian siRNA expression vector pUC18U6. Then oligonucleotides coding for a short hairpin RNA against HBV were cloned into pUC18U6 to form pUC18U6HBVsir which was introduced into 2.2.15 cells by using liposome-mediated transfection. 2.2.15 cells transfected by pUC18U6 and pUC18U6GFPsir which expressed siRNA against green fluorescent protein and mock-transfected 2.2.15 cells were used as controls. Concentration of HBsAg in the supernatant of the transfected cells was measured by using solid-phase radioimmunoassay. RESULTS: A mammalian siRNA expression vector pUC18U6 was constructed successfully. Compared with controls, pUC18U6HBVsir which expressed siRNA against HBV decreased concentration of HBsAg significantly by 44%(P<0.05). CONCLUSION: HBV replication in 2.2.15 cells is inhibited by siRNA expressed from the DNA vector.

Targeted ribonuclease can inhibit replication of hepatitis B virus.

AIM: To study the effect of a novel targeted ribonuclease (TN), the fusion protein of HBVc and human eosinophil-derived neurotoxin (hEDN), on the HBV replication in vitro. METHODS: The gene encoding the targeted ribonuclease was cloned into pcDNA3.1 (-) to form recombinant eukaryotic expression vector p/TN. Control plasmids, including p/hEDN, p/HBVc, and p/TNmut in which a Lys113-Arg mutation was introduced by sequential PCR to eliminate the ribonuclease activity of hEDN, were also constructed. Liposome-mediated transfection of 2.2.15 cells by p/TN, p/TNmut, p/hEDN, p/HBVc, and pcDNA3.1 (-), or mock transfection was performed. After that, RT-PCR was used to verify the transgene expression. Morphology of the transfected cells was observed and MTT assay was performed to detect the cytotoxicity of transgene expression. Concentration of HBsAg in the supernatant of the transfected cells was measured using solid-phase radioimmunoassay. RESULTS: Transgenes were successfully expressed in 2.2.15 cells. No obvious cytotoxic effect of transgene expression on 2.2.15 cells was found. The HBsAg concentration in the p/TN transfected cells was reduced by 58 % compared with that of mock transfected cells. No such an effect was found in all other controls. CONCLUSION: The targeted ribonuclease can inhibit HBV replication in vitro while it has no cytotoxicity on host cells. The targeted ribonuclease may be used as a novel antiviral agent for human HBV infection.