OBJECTIVES: To study the molecular mechanisms of LDH-loaded si-NEAT1 for regulating paclitaxel resistance and tumor-associated macrophage (TAM) polarization in breast cancer. METHODS: qRT-PCR and Western blotting were used to detect the expression of lncRNA NEAT1, miR-133b, and PD-L1 in breast cancer SKBR3 cells and paclitaxel-resistant SKBR3 cells (SKBR3-PR). The effects of transfection with si-NEAT1 and miR-133b mimics on MRP, MCRP and PD-L1 expressions and cell proliferation, migration and apoptosis were investigated using qRT-PCR, Western blotting, scratch and Transwell assays, and flow cytometry. Rescue experiments were conducted using si-NEAT1 and miR-133b inhibitor. Human THP-1 macrophages were cultured in the presence of conditioned media (CM) derived from SKBR3 and SKBR3-PR cells with or with si-NEAT1 transfection for comparison of IL-4-induced macrophage polarization by detecting the surface markers. LDH@si-NEAT1 nanocarriers were constructed, and their effects on MRP, MCRP and PD-L1 expressions and cell behaviors of the tumor cells were examined. THP-1 cells were treated with the CM from LDH@si-NEAT1-treated tumor cells, and the changes in their polarization were assessed. RESULTS: SKBR3-PR cells showered significantly upregulated NEAT1 and PD-L1 expressions and lowered miR-133b expression as compared with their parental cells. Transfection with si-NEAT1 and miR-133b mimics inhibited viability, promoted apoptosis and enhanced MRP and BCRP expressions in SKBR3-PR cells. NEAT1 knockdown obvious upregulated miR-133b and downregulated PD-L1, MRP and BCRP expressions. The CM from SKBR3-PR cells obviously promoted M2 polarization of THP-1 macrophages, which was significantly inhibited by CM from si-NEAT1-transfected cells. Treatment with LDH@si-NEAT1 effectively inhibited migration and invasion, promoted apoptosis, and reduced MRP, BCRP and PD-L1 expressions in the tumor cells. The CM from LDH@si-NEAT1-treated SKBR3-PR cells significantly downregulated Arg-1, CD163, IL-10, and PD-L1 and upregulated miR-133b expression in THP-1 macrophages. CONCLUSIONS LDH@si-NEAT1 reduces paclitaxel resistance of breast cancer cells and inhibits TAM polarization by targeting the miR-133b/PD-L1 axis.
Humans ; MicroRNAs/genetics* ; RNA, Long Noncoding/genetics* ; Paclitaxel/pharmacology* ; Breast Neoplasms/metabolism* ; Drug Resistance, Neoplasm ; B7-H1 Antigen/metabolism* ; Cell Line, Tumor ; Female ; Tumor-Associated Macrophages ; Apoptosis ; Cell Proliferation ; Macrophages ; Cell Movement

Objective:To explore the impact of early and late antibody-mediated rejection (AMR) on treatment options and allograft outcomes after kidney transplantation (KT).Methods:From January 2013 to December 2022, the study retrospectively enrolled 141 KT allograft recipients receiving allograft biopsy and diagnosed as AMR according to the Banff 2019 criteria. Recipients with a diagnosis of AMR within 30 days post-KT were classified into early AMR group (n=19) while the remainders assigned as late AMR group (n=122). The outcome endpoints included recipient survival rate, death-censored graft survival rate, follow-up estimated glomerular filtration rate (eGFR) and immunodominant donor-specific antibody (DSA) intensity. Wilcoxon's test was utilized for assessing the differences in eGFR and DSA intensity while Kaplan-Meier curve and Log-rank test were employed for evaluating graft survival impact. Treatment regimens for AMR were collected and categorized.Results:The median follow-up duration was 2.6(1.2, 5.2) year. No graft failure was noted in early AMR group while 44 recipients in late AMR group experienced graft failure, with 34 cases (77.2%) due to AMR progression. The 5-year death-censored graft survival rate was significantly better in early AMR group than that in late AMR group [100% vs 60.1%(50.5%, 71.6%), P=0.002]. The one-year change in eGFR for early AMR group was significantly superior to that of late AMR group [19.3(-2.6, 38.1) vs -3.3(-14.0, 5.4), P=0.001]. One-year mean fluorescent intensity (MFI) of early AMR group was 1 158(401.5, 3 126.5). It was significantly lower than that when diagnosed with early AMR [3 120.5(2 392.8, 9 340.0)] and one-year MFI of late AMR group [8 094(2 251.5, 13 560.5)] ( P=0.005, P<0.001). Early AMR group primarily received standard treatment (3/19, 15.8%) and regimens centered on rituximab and/or bortezomib (7/19, 43.8%). Late AMR group mainly received standard (16/122, 13.1%) or intensified regimens (9/122, 7.4%) and regimens focused upon rituximab and/or bortezomib (32/122, 26.2%) and MP monotherapy (21/122, 17.2%). Conclusion:The outcome for early AMR is significantly better than that for late AMR. For early AMR, early and robust immunosuppression is recommended. For late AMR, early detection and timely treatment are crucial and individualized strategies should be implemented.

Although sympathetic blockade is clinically used to treat pain, the underlying mechanisms remain unclear. We developed a localized microsympathectomy (mSYMPX), by cutting the grey rami entering the spinal nerves near the rodent lumbar dorsal root ganglia (DRG). In a chemotherapy-induced peripheral neuropathy model, mSYMPX attenuated pain behaviors via DRG macrophages and the anti-inflammatory actions of transforming growth factor-β (TGF-β) and its receptor TGF-βR1. Here, we examined the role of TGF-β in sympathetic-mediated radiculopathy produced by local inflammation of the DRG (LID). Mice showed mechanical hypersensitivity and transcriptional and protein upregulation of TGF-β1 and TGF-βR1 three days after LID. Microsympathectomy prevented mechanical hypersensitivity and further upregulated Tgfb1 and Tgfbr1. Intrathecal delivery of TGF-β1 rapidly relieved the LID-induced mechanical hypersensitivity, and TGF-βR1 antagonists rapidly unmasked the mechanical hypersensitivity after LID+mSYMPX. In situ hybridization showed that Tgfb1 was largely expressed in DRG macrophages, and Tgfbr1 in neurons. We suggest that TGF-β signaling is a general underlying mechanism of local sympathetic blockade.
Mice ; Animals ; Receptor, Transforming Growth Factor-beta Type I/metabolism* ; Transforming Growth Factor beta/pharmacology* ; Transforming Growth Factor beta1/metabolism* ; Hyperalgesia/metabolism* ; Radiculopathy/metabolism* ; Pain/metabolism* ; Analgesics/pharmacology* ; Ganglia, Spinal/metabolism*

Objective:To investigate the clinical value of peripheral monocyte and neutrophil count in predicting the response of patients with metastatic non-small cell lung cancer (mNSCLC) to immunosuppressive checkpoint inhibitors (ICI).Methods:The clinical data of 34 adult mNSCLC patients who received nafulizumab or pabolizumab in Danzhou People's Hospital of Hainan Province from January 2017 to March 2019 were retrospectively analyzed. The correlation of the demographic characteristics, clinical data, hematological examination results in the first two weeks before the treatment and two weeks after ICI treatment with prognosis was recorded and observed.Results:The baseline mean monocyte count [(0.52±0.09)×10 9/L vs. (0.60±0.12)×10 9/L] and neutrophil count [(4.27±0.87)×10 9/L vs.(5.39±1.02)×10 9/L] of patients with ICI reaction were lower than those of patients without ICI reaction, and the differences were statistically different ( t = -2.572, -2.727, all P < 0.05). However, there was a negative correlation between the monocyte count of the patients who responded to ICI and the reaction time ( r = -0.507, P < 0.05). The median reaction time in patients with monocyte count >0.70×10 9/L was shorter than that in patients with monocyte count ≤0.70×10 9/L (8 weeks vs. 12 weeks, χ2=4.162, P = 0.041). There was no correlation between monocyte count and time of reaction duration, progression of free survival (PFS) and overall survival (OS) ( r = -0.214, 0.182, 0.232, all P > 0.05). The decrease rate of neutrophil count in response group was higher than that in non-response group (22% vs. 2%, P < 0.05). After the first administration, cutoff value of neutrophil count was 4.2×10 9/L; the response rate of patients with neutrophil count ≤ 4.2×10 9/L was higher than that of patients with neutrophil count > 4.2×10 9/L [86.7% (13/15) vs. 36.8% (7/19), χ2=6.657, P < 0.05]. Conclusion:Peripheral blood monocyte and neutrophil count can predict the response to ICI therapy in patients with mNSCLC.

Objective:The recombinant shuttle peptide-single chain antibody with neutralizing activity is expected to be obtained by combining the shuttle peptide sequence with rabies single chain antibody and optimizing the expression conditions in E. coli.Methods:In this study, the SO57 single-stranded antibody sequence was recoded and the shuttle peptide of Arg 12 was fused at the N-terminal to construct pET22 (b)-rSO57 expression vector, which was then expressed in E. coli. The growth density OD 600, induction time, induction temperature and inducer concentration were optimized to obtain a higher expression effect. The recombinant protein was purified by immobilized metal chelating chromatography, and the relative affinity of the recombinant single-chain variable fragment (scFv) was determined. The neutralization effect of the recombinant scFv was verified by rSO57/CVS-11 virus strain mixed with infected cells and rapid fluorescent focus inhibition test (RFFIT). Results:The recombinant rSO57 was successfully constructed and induced in BL21 (de3). The optimal expression condition was that when the cell density of OD 600 was 0.8, IPTG of 0.4 mmol/ml was used for induction. After induction at 16 ℃ for 24 hours, rSO57 was expressed in soluble form, accounting for 19.8% of the soluble protein in the cell. After chromatography purification, the recombinant rSO57 with a purity of 84% was obtained. ELISA showed that the relative affinity coefficient Ka of rSO57 was 4.3×10 5. When rSO57 was mixed with CVS-11 strain, the infection rate of cells increased with the increase of dilution, indicating the neutralization activity of recombinant antibodies. Using the RFFIT, 50 μg rSO57 was equivalent to 2.17 IU of rabies neutralizing serum. Conclusions:In this study, the recombinant scFv fusion with shuttle peptide was expressed, which could neutralize rabies virus, in order to prepare specific and targeted antiviral drug carriers for rabies treatment.

Objective:To assess the application value of China consensus on the protocol of early gastric cancer screening in Guangdong province.Methods:A new quantitative scoring system was used in Cantonese residents who underwent early gastric cancer screening from March 2018 to March 2019. According to the scores of initial screening, patients were divided into high-risk, medium-risk and low-risk groups. The detection rates of early gastric cancer, precancerous diseases and precancerous lesions under gastroscopy in each group were compared. Chi-square test was performed for statistical analysis.Results:A total of 545 individuals were selected for gastroscopy, in which 32 cases were classified into high-risk group, 184 into medium-risk group and 329 into low-risk group. The results of gastroscopy examination showed that high-risk group had the highest detection rate of early gastric cancer (12.5%), followed by medium-risk group (1.1%) and low-risk group (0) ( χ2=41.85, P<0.01); the detection rates of precancerous diseases exhibited a similar pattern: high-risk group (60.9%) > medium-risk group (52.4%) > low-risk group (34.3%) ( χ2=18.00, P<0.01). The detection rates of precancerous lesions were 17.9%, 8.8% and 8.8%, respectively, with no significant difference ( χ2=2.58, P=0.28). In terms of the positive rate of endoscopy, high-risk group (71.9%) showed the highest positive rate, followed by medium-risk group (57.1%) and low-risk group (40.1%) ( χ2=21.54, P<0.01). Conclusion:China consensus on the protocol of early gastric cancer screeing is of application value for the screening of early gastric cancer and precancerous lesions in the populations at risk of gastric cancer in Guangdong province.

Objective:To systematically evaluate the incidence of coronavirus disease 2019 (COVID-19) syndrome in traditional Chinese medicine (TCM), and to provide a basis for the standard of COVID-19 syndrome differentiation.Methods:CNKI, China Medical Journal Network, PubMed and Embase databases were searched by computer, and TCM syndrome data of COVID-19 cross-sectional surveys and case series were collected from the establishment of the database to March 31st in 2020. Two researchers independently screened the literature, extracted relevant data such as TCM syndromes, and evaluated the quality of the literature according to the quality assessment tool recommended by the JBI Evidence-Based Health Care Center. Stata 14.0 software was used to perform a single-group Meta-analysis of TCM syndromes.Results:Thirteen cross-sectional studies and 5 case series studies were included, with a total of 2 139 patients. The analysis of the cross-sectional studies showed that the risk bias was higher in the identification, control and accuracy of the confounding factors, and the description of the reliability and validity of the outcome indicators was unclear. The analysis of the case series studies showed that the overall risk bias was small, but there was a lack of a comprehensive description of the assessment when selecting patients. A total of 33 TCM syndromes of COVID-19 were sorted out by combining the TCM syndromes with the same disease position and pathogenic syndrome in the included 18 literatures, and there were 10 types of TCM syndromes of which the number of occurrences was ≥50, the incidence rate was ≥10%, and had statistical significance [indicated that 95% confidence interval (95% CI) didn't cross the meaningless line 0], including cold dampness syndrome (469 cases, incidence rate was 28.2%, 95% CI was 5.9%-50.6%), damp heat syndrome (247 cases, incidence rate was 32.1%, 95% CI was 10.7%-53.6%), epidemic closed lung syndrome (228 cases, incidence rate was 28.9%, 95% CI was 12.5%-45.2%), pulmonary and spleen Qi deficiency syndrome (202 cases, incidence rate was 22.1%, 95% CI was 9.5%-34.7%), dampness stagnation lung syndrome (180 cases, incidence rate was 41.2%, 95% CI was 18.4%-64.1%), dampness obstructing lung and spleen syndrome (81 cases, incidence rate was 56.3%, 95% CI was 48.1%-64.4%), evil heat syndrome lung syndrome (76 cases, incidence rate was 31.1%, 95% CI was 25.3%-36.9%), dampness-blocking lung and stomach syndrome (70 cases, incidence rate was 13.4%, 95% CI was 10.4%-16.3%), heat poisoning lung closure syndrome (55 cases, incidence rate was 16.9%, 95% CI was 8.3%-25.5%), and Qi-Yin deficiency syndrome (53 cases, incidence rate was 13.7%, 95% CI was 2.7%-24.8%). Subgroup analysis showed that there were 6 types of TCM syndromes that met the above conditions in cross-sectional studies, namely cold and damp lung syndrome (200 cases, incidence rate was 20.9%, 95% CI was 12.6%-29.2%), damp heat syndrome (221 cases, incidence rate was 41.8%, 95% CI was 9.4%-74.2%), dampness-disease lung syndrome (120 cases, incidence rate was 41.4%, 95% CI was 8.6%-74.3%), lung and spleen Qi deficiency syndrome (115 cases, incidence rate was 19.2%, 95% CI was 6.8%-31.7%), heat-fever lung syndrome (76 cases, incidence rate was 31.1%, 95% CI was 25.3%-36.9%) and Qi-Yin deficiency syndrome (53 cases, incidence rate was 13.7%, 95% CI was 2.7%-24.8%). There were 3 kinds of TCM syndromes in case series studies, which were virus closed lung syndrome (133 cases, incidence rate was 44.0%, 95% CI was 24.5%-63.4%), lung and spleen Qi deficiency syndrome (87 cases, incidence rate was 38.7%, 95% CI was 32.3%-45.0%), and dampness and depression lung syndrome (60 cases, incidence rate was 40.6%, 95% CI was 29.3%-52.0%). Conclusions:The TCM syndromes of COVID-19 syndrome were widely distributed and complex. The main TCM syndromes were cold dampness syndrome, damp heat syndrome, epidemic closed lung syndrome, pulmonary and spleen Qi deficiency syndrome, dampness stagnation lung syndrome, dampness obstructing lung and spleen syndrome, evil heat syndrome lung syndrome, dampness-blocking lung and stomach syndrome, heat poisoning lung closure syndrome, Qi-Yin deficiency syndrome, which can provide reference for the standard of TCM syndrome differentiation of COVID-19.

Objective:To explore the correlation between symptoms and their contribution to syndrome based on syndrome of lung damp-heat accumulation in coronavirus disease 2019 (COVID-19), thus to provide methodological basis for the syndrome diagnosis.Methods:Based on 654 clinical investigation questionnaires data of COVID-19 patients, a model based on syndrome of lung damp-heat accumulation was set. Using SPSS Modeler 14.1 software, association rules and Bayesian network were applied to explore the correlation between symptoms and their contribution to syndrome.Results:There were 121 questionnaires referring to syndrome of lung damp-heat accumulation in total 654 questionnaires. The symptoms with frequency > 40% were fever (53.72%), cough (47.93%), red tongue (45.45%), rapid pulse (43.80%), greasy fur (42.15%), yellow tongue (41.32%), fatigue (40.50%) and anorexia (40.50%). Association rule analysis showed that the symptom groups with strong binomial correlation included fever, thirst, chest tightness, shortness of breath, cough, yellow phlegm, etc. The symptom groups with strong trinomial correlation included cough, yellow phlegm, phlegm sticky, anorexia, vomiting, heavy head and body, fever, thirst, fatigue, etc. Based on SPSS Modeler 14.1 software, with syndrome of lung damp-heat accumulation (yes = 1, no = 0) as target variable, and the selected symptoms with frequency > 15.0% as input variables, the Bayesian network model was established to obtain the probability distribution table of symptoms (groups), in which there was only one parent node (the upper node of each input variable) of fever, and the conditional probability was 0.54. The parent node of cough had yellow phlegm and syndrome of lung damp-heat accumulation, indicating that there was a direct causal relationship between cough and yellow phlegm in syndrome of lung damp-heat accumulation, and the conditional probability of cough was 0.99 under the condition of yellow phlegm. The common symptom groups and their contribution to syndrome were as follows: fever and thirsty (0.47), cough and yellow phlegm (0.49), chest tightness and polypnea (0.46), anorexia and heavy cumbersome head and body (0.61), yellow greasy fur and slippery rapid pulse (0.95).Conclusions:It is feasible and objective to analyze the correlation between symptoms and their contribution to syndromes by association rules combined with Bayesian network. It could provide methodological basis for the syndrome diagnosis.

Objective To summarize the outcomes and clinical experiences of renal transplantation in human immunodeficiency virus (HIV)-positive patients .Methods The clinical data were retrospectively analyzed for one HIV-positive case of renal transplantation .Diagnosed as chronic renal insufficiency 1 year ago ,he received hemodialysis .After a positive screen for HIV ,he received highly active antiretroviral therapy (HAART) and HIV RNA turned negative 3 months later .CD4 + T cell count was 331 cell/μl at pre-operation and there was no HIV-rated opportunistic infection or cancer . Her mother donated her kidney . Basiliximab and steroid pulse therapy were used preoperatively and immunosuppressants were used after transplantation , including tacrolimus , corticosteroids and mycophenolate mofetil .Results The kidney was transplanted successfully and serum creatinine declined to a normal level at day 4 after transplantation .Because of an interaction between efaverenz and tacrolimus ,the blood concentration of tacrolimus was extremely low and the dose of tacrolimus had to be raised to 0 .2 mg/(kg·d) .Antiroviral therapy remained unchanged .No rejection and other complications were observed .And HIV RNA remained negative .Conclusions Renal transplantation is optimal for HIV-positive patients whose HIV status is completely under control .However ,drug interactions needs to be considered during perioperative and postoperative periods .

Injury to peripheral nerves can lead to neuropathic pain, along with well-studied effects on sensory neurons, including hyperexcitability, abnormal spontaneous activity, and neuroinflammation in the sensory ganglia. Neuropathic pain can be enhanced by sympathetic activity. Peripheral nerve injury may also damage sympathetic axons or expose them to an inflammatory environment. In this study, we examined the lumbar sympathetic ganglion responses to two rat pain models: ligation of the L5 spinal nerve, and local inflammation of the L5 dorsal root ganglion (DRG), which does not involve axotomy. Both models resulted in neuroinflammatory changes in the sympathetic ganglia, as indicated by macrophage responses, satellite glia activation, and increased numbers of T cells, along with very modest increases in sympathetic neuron excitability (but not spontaneous activity) measured in ex vivo recordings. The spinal nerve ligation model generally caused larger responses than DRG inflammation. Plasticity of the sympathetic system should be recognized in studies of sympathetic effects on pain.
Action Potentials ; physiology ; Animals ; Disease Models, Animal ; Female ; Ganglia, Sympathetic ; pathology ; Glial Fibrillary Acidic Protein ; metabolism ; Hyperalgesia ; etiology ; Ligation ; adverse effects ; Macrophages ; pathology ; Male ; Neurogenic Inflammation ; etiology ; Pain ; etiology ; pathology ; Patch-Clamp Techniques ; Peripheral Nerve Injuries ; complications ; Rats ; Rats, Sprague-Dawley ; Receptors, Antigen, T-Cell, alpha-beta ; metabolism