Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by synovial inflammation, joint destruction, and functional impairment. Angiogenesis plays a key role in the pathological progression of RA with dysfunction of endothelial cells to promote synovial inflammation, sustain pannus formation, subsequently leading to joint damage. Colquhounia Root Tablets (CRT), a Chinese patent drug, has shown a satisfying clinical efficacy in treating RA, while the underlying mechanism by which CRT inhibits RA-associated angiogenesis remains unclear. In this study, we applied a research approach combining transcriptomic data analysis, bio-network mapping, and in vivo and in vitro experiments to explore the molecular mechanisms of CRT in suppressing angiogenesis in RA. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of Institute of Basic Theory for Chinese Medicine, China Academy of Chinese Medical Sciences (ratification number: IBTCMCACMS21-2307-06). Network analysis identified that key genes such as nucleotide-binding oligomerization domain-containing protein 2 (NOD2), SMAD family member 3 (SMAD3), and vascular endothelial growth factor A (VEGFA) significantly enriched in pathways related to NOD-like receptor signaling and VEGF signaling, indicating that CRT may inhibit angiogenesis by regulating vascular endothelial cell function with modulating angiogenesis-related pathways. In vivo data showed that CRT significantly reduced the positive expression of CD31 and VEGF in the ankle joint of adjuvant-induced arthritis (AIA) rats. In vitro data further confirmed that CRT effectively inhibited VEGF-induced migration, invasion, and tube formation in HUVECs, while significantly reduced the expression of angiogenesis-related factors VEGF/CD31/Ang-1, as well as the positive expression of VEGF and CD31 in HUVECs. Furthermore, CRT markedly decreased the protein expression of NOD2, VEGFA, and SMAD3. In conclusion, these findings indicate that CRT may inhibit the RA-related angiogenesis by targeting the NOD2/SMAD3/VEGF signaling axis to improve endothelial cell function, enriching the scientific connotation of CRT in inhibiting pathological angiogenesis in RA and also offer new insights for clinical prevention and treatment of RA.

: AbstractObjective: To investigate the effect of γδ T cells on the proliferation, apoptosis and autophagy of multiple myeloma cells. METHODS: Peripheral blood mononuclear cells (PBMNC) were isolated from healthy volunteers, and stimulated with zoledronic acid (Zol) in combination with rhIL-2. Flow cytometry analysis was used to detected the purity of γδ T cells. γδ T cells were collected and co-cultured with RPMI-8226 or U-266 cells at different effector target ratios. The proliferation of RPMI-8226 or U-266 cell lines were detected by CCK-8. Cell cycle and cell apoptosis were detected by flow cytometry and Western blot．The expressions of autophagy-related proteins were detected by Western blot. RESULTS: γδ T cells can be expanded in vitro. γδ T cells could inhibit the proliferation of RPMI-8226 or U-266 cells, induced cell cycle arrest and promoted apoptosis in an effector target-dependent manner. In addition, γδ T cells could induce autophagy of myeloma cells, inhibited the expression of autophagy-related PI3K, P-AKT and P-mTOR, while increased the expression of AMPK and Beclin-1. CONCLUSION γδ T cells can inhibit the proliferation of RPMI-8226 and U-266 myeloma cells, induce cell cycle arrest, promote apoptosis, and enhance autophagy in vitro. The mechanism may be related to inhibition of PI3K/AKT/mTOR signaling pathway and/or activation of AMPK/Beclin-1 signaling pathway.
AMP-Activated Protein Kinases/pharmacology* ; Apoptosis ; Autophagy ; Beclin-1/pharmacology* ; Cell Proliferation ; Humans ; Leukocytes, Mononuclear/metabolism* ; Multiple Myeloma/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; T-Lymphocytes ; TOR Serine-Threonine Kinases/metabolism*

This study investigated the anti-ascites effect of the total saponins of Phytolaccae Radix(PRTS) and the mechanism.H22 cell suspension was used(ip) to induce ascites in ICR male mice, and the model mice were randomized into model group, positive drug group(furosemide, 6 mg·kg~(-1)), total extract of Phytolaccae Radix(PRTE) group, and PRTS(1.29 g·kg~(-1)).Another 10 male mice were selected as the blank group.Mice in the blank group and model group were given(ig) normal saline containing 0.5% CMC-Na, and those in the positive drug group, PRTE group, and PRTS group received(ig) corresponding doses of drugs, once a day, for 8 consecutive days.The ascites volume, urine volume, and fecal water content in mice with ascites, serum levels of antidiure-tic hormone(ADH), renin in renin-angiotensin-aldosterone system(RAAS), angiotensin Ⅱ(AngⅡ), and aldosterone(ALD), expression of aquaporin(AQP)1-AQP4 in kidney, expression of AQP1, AQP3 in colon, and expression of phosphatidylinositol 3-kinase/protein kinase B(PI3 K/Akt) pathway-related proteins were detected to explore the anti-ascites mechanism of PRTS.The results showed that the PRTS can increase the urine volume and fecal water content and decrease the ascites volume of ascites mice.Moreover, PRTS significantly reduced the expression of AQP1-AQP4 in kidney and AQP1, AQP3 in colon, serum levels of renin, AngⅡ, ALD, and ADH, and the expression of p-PI3 K and p-Akt in the kidney of ascites mice.PRTS exerts anti-ascites effect by promoting urination and defecation.The mechanism is that it inhibits the activities of RAAS and ADH and suppresses the phosphorylation of PI3 K/Akt signaling pathway, thereby restricting the expression of AQPs in the kidney and colon.
Animals ; Aquaporin 1 ; Ascites/metabolism* ; Male ; Mice ; Mice, Inbred ICR ; Proto-Oncogene Proteins c-akt/metabolism* ; Renin/metabolism* ; Saponins/pharmacology* ; Water/metabolism*

The present study aims to investigate the correlation between irritant toxicity variation and lectin content variation during the processing of Pinelliae Rhizoma products and to explore the feasibility of Western blot as a method for the detection of lectin. We processed Pinelliae Rhizoma Praeparatum Cum Alumine, Pinelliae Rhizoma Praeparatum, and Pinelliae Rhizoma Praeparatumcum Zingibere et Alumine to different degrees and then analyzed their irritant toxicity via Draize rabbit eye test. Western blot was employed to determine the lectin content in Pinelliae Rhizoma products processed with different methods. The correlation between toxicity variation and lectin content variation was then analyzed. Different decoction pieces of Pinelliae Rhizoma were collected for the determination of lectin content. The three processed products of Pinelliae Rhizoma showed gradually decreased toxicity and lectin content as the processing continued. The decreasing trend of lectin content was consistent with that of irritant toxicity during processing, which indicated that the change in lectin content could reflect the trend of irritant toxicity. No band of lectin appeared in the Western blot of processed products of Pinelliae Rhizoma, which suggested that western blotting can be used for the detection of toxic lectin in the processed products of Pinelliae Rhizoma. Lectin should not be detected in the Pinelliae Rhizoma products processed according to the methods in the Chinese Pharmacopoeia.
Animals ; Drugs, Chinese Herbal/toxicity* ; Irritants ; Lectins ; Pinellia ; Rabbits ; Technology, Pharmaceutical/methods*

To determine the content of endogenous toxic substance Pinellia ternata lectin(PTL) protein in Pinelliae Rhizoma and the related processed products, this study prepared specific monoclonal antibodies against PTL by hybridoma cell technology, and established a quantitative double-antibody sandwich enzyme linked immunosorbent assay(ELISA) for PTL antigen. The detection conditions were 2.5 μg·mL~(-1) working concentration of the captured antibody and 1∶450 of the dilution multiple of detected antibody. The coating condition was staying overnight at 4 ℃. The blocking time and incubation times of antigen and detected antibody were all 90 minutes. The incubation time of horseradish peroxidase conjugated streptavidin-horseradish peroxidase(SA-HRP) was 15 minutes. The quantitative limit of the method for PTL antigen was 0.375 ng·mL~(-1). The linear range was 75.000-4 800.000 pg·mL~(-1), and R~2=0.997 1. The recovery rate was 90.0%-110.0%, and the variation coefficients of intra-test and inter-test precision were 2.0%-3.0% and 2.0%-8.5%.The content of PTL in three batches of Pinelliae Rhizoma and the related processed products was determined by the method, and the average content of PTL in Pinelliae Rhizoma was 35.42 mg·g~(-1). The average content of PTL in Pinelliae Rhizoma Praeparatum Cum Alumine, Pinelliae Rhizoma Praeparatum, and Pinelliae Rhizoma Praeparatum Cum Zingibere Et Alumine were 1.15 mg·g~(-1), 16.53 μg·g~(-1), and 122.63 ng·g~(-1), respectively, indicating that the content of PTL decreased significantly after processing. The quantitative double-antibody sandwich ELISA for PTL antigen established in this paper had good linearity, sensitive response, and high accuracy, which provided a simple and effective monitoring method for the detection of PTL content in the processing of Pinelliae Rhizoma.
Pinellia ; Drugs, Chinese Herbal ; Antibodies, Monoclonal ; Enzyme-Linked Immunosorbent Assay ; Horseradish Peroxidase

Objective:To study the clinical characteristics of children with adenovirus pneumonia complicated with liver injury and the understanding of clinicians.Methods:The children diagnosed with community-acquired adenovirus pneumonia were enrolled from January 1, 2013 to December 31, 2020. The clinical characteristics of the normal liver function group and the liver injured group were compared, and multivariate regression was applied.Results:Among the 3 294 enrolled children, 1 704 children had elevated alanine aminotransferase and/or aspartate aminotransferase on admission, and the liver function impairment rate was 51.7% (1 704/3 294), of which 1 671 cases were mildly injured (98.06%, 1 671/1 704), 28 cases were moderately injured (1.64%, 28/1 704), and 5 cases were severely injured (0.29%, 5/1 704). Compared with the normal liver function group, the proportion of male children in the liver injured group was higher (68.1% vs. 64.2%, P=0.017), and the age was younger[16(9, 30) months vs. 38(19, 53) months, P<0.001]. Children with liver injury were more likely to have wheezing (32.9% vs. 18.7%, P<0.001), shortness of breath (13.7% vs. 9.1%, P<0.001), and diarrhea (11.0% vs. 4.7%, P<0.001). The white blood cell counts, neutrophil counts and hemoglobin in the liver injured group were lower than those in the normal group, while the lymphocyte counts were higher than those in the normal group. The levels of creatine kinase, cardiac creatine kinase and lactate dehydrogenase were higher. The proportion of severe pneumonia in the liver injured group (26.9% vs. 11.2%, P<0.001) was higher than that in the normal liver function group, and the case fatality rate was significantly higher (1.47% vs. 0.25%, P<0.001). Comparing the probability of liver injury in children with mild and severe adenovirus pneumonia, it was found that the severe pneumonia group had more frequent liver injury (73.0% vs 46.6%, P<0.001), and both moderate and severe liver injury groups were higher than in the mild group. Further multivariate logistic regression analysis did not find a significant factor associated with the incidence of liver injury. Conclusions:The incidence of adenovirus pneumonia in children with liver injury was high, and mild injury is more common. Children with liver injury were younger, had more severe clinical symptoms such as wheezing and shortness of breath, and higher proportion of severe pneumonia and mortality. Among them, children with severe pneumonia had a higher proportion of liver injury, and moderate to severe injury was more common. The monitoring of children with different degrees of liver injury should be strengthened.

This paper reports 3 cases of acute hepatitis of unknown cause in children who met the case definition of WHO. Human adenovirus group C was detected in case 3, and the quantity of viral DNA was relatively high, which may be related to the liver function damage in the patient, but its role in pathogenesis needs further study to confirm.

Objective:To investigate the correlations of human leukocyte antigen (HLA)-A, B, C, and DRB1 genotypes with cross-reactivity caused by aromatic antiepileptic-drugs.Methods:A case-control association study was carried out on subjects who accepted treatments/physical examination in our hospitals from September 2016 and September 2020; 31 patients with aromatic antiepileptic drugs (carbamazepine, phenytoin, oxcarbazepine, lamotrigine and phenobarbital)-induced cross-reactivity were enrolled as patient group, 52 tolerant subjects who took the 5 antiepileptic drugs for more than 3 months without cross-reactivity were chosen as tolerant control group, and 500 healthy volunteers were recruited as normal control group. The ethnicity of all patients and controls was Han Chinese. High-resolution genotyping was performed to compare the HLA-A, B, C, and DRB1 genotypes in subjects of the 3 groups. χ2 test or Fisher's exact test were used to analyze the correlations of HLA genes with cross-reactivity caused by aromatic antiepileptic-drugs. Results:The presence of HLA-B*13:01 genotype in the patient group, the tolerant control group, and the normal control group was 45.2% (14/31), 15.4% (8/52) and 14.6% (73/500), respectively. The presence of HLA-B*13:01 genotype in the patient group was significantly higher as compared with that in the tolerant control group and normal control group ( Pc<0.017). No other HLA genotypes were found to be associated with cross-reactivity caused by aromatic antiepileptic-drugs. Conclusion:HLA-B*13:01 is the risk genotype for cross-reactivity caused by aromatic antiepileptic-drugs.

Objective: To analyze the clinical characteristics of fatal cases with confirmed severe adenovirus pneumonia in children in order to improve the diagnosis and treatment. Methods: The fatal cases of severe adenovirus pneumonia admitted to Pediatric Intensive Care Unit of Hunan Provincial People′s Hospital from January 2019 to July 2019 were collected, whose clinical features, diagnosis, treatment, and the causes of death were analyzed retrospectively. Results: A total of eight children were enrolled, and the age ranged from 3 months to 3 years old, and five cases were between 6 months to 2 years old.Three cases had underlying diseases.Adenovirus genotype identification was performed on six patients, and the results showed that all patients were infected with adenovirus type 7.All patients presented persistent high fever, with a peak temperature between 39.8℃ to 41.0℃, which persisted 10 to 37 days.Blood routine test before admission PICU showed that four cases had the decrease in white blood count and hemoglobin concentration, accompanied byincreased serum ferritin levels.Seven cases complicated with infection.Four cases had parainfluenza virus type 3 infection.Six cases had bacterial infection, and Gram-negative bacilli were predominant.One had fungal septicemia.Conventional mechanical ventilation were performed in all patients in this group.Four cases in this group died of severe acute respiratory distress syndrome.The other four cases died of disseminated intravascular coagulation associated with massive gastrointestinal bleeding, pulmonary hemorrhage, heart failure and septic shock combined with multiple organ failure. Conclusion Fatal adenoviruspneumonia mostly apppears in children between 6 months to 2 years old or with underlying diseases.Adenovirus type 7 was the main serotype.The occurrence of reactive hemophagocytic phenomenon should be worsen progression of the disease.Co-infection may be an important cause of death.

Objective To analyze the clinical characteristics of fatal cases with confirmed severe ad﹣enovirus pneumonia in children in order to improve the diagnosis and treatment. Methods The fatal cases of severe adenovirus pneumonia admitted to Pediatric Intensive Care Unit of Hunan Provincial People′s Hospital from January 2019 to July 2019 were collected,whose clinical features,diagnosis,treatment,and the causes of death were analyzed retrospectively. Results A total of eight children were enrolled, and the age ranged from 3 months to 3 years old,and five cases were between 6 months to 2 years old. Three cases had underly﹣ing diseases. Adenovirus genotype identification was performed on six patients,and the results showed that all patients were infected with adenovirus type 7. All patients presented persistent high fever,with a peak temper﹣ature between 39. 8℃ to 41. 0℃,which persisted 10 to 37 days. Blood routine test before admission PICU showed that four cases had the decrease in white blood count and hemoglobin concentration,accompanied by﹣increased serum ferritin levels. Seven cases complicated with infection. Four cases had parainfluenza virus type 3 infection. Six cases had bacterial infection,and Gram﹣negative bacilli were predominant. One had fun﹣gal septicemia. Conventional mechanical ventilation were performed in all patients in this group. Four cases in this group died of severe acute respiratory distress syndrome. The other four cases died of disseminated intra﹣vascular coagulation associated with massive gastrointestinal bleeding, pulmonary hemorrhage, heart failure and septic shock combined with multiple organ failure. Conclusion Fatal adenoviruspneumonia mostly app﹣pears in children between 6 months to 2 years old or with underlying diseases. Adenovirus type 7 was the main serotype. The occurrence of reactive hemophagocytic phenomenon should be worsen progression of the disease. Co﹣infection may be an important cause of death.