This study investigated the effects and action mechanism of tannins from Galla chinensis cream(TGCC) on the skin wound of rat tail. Male Sprague Dawley(SD) rats were randomly divided into a control group, model group, model+low-dose TGCC(50 mg per rat) group, model+high-dose TGCC group(100 mg per rat), and model+TGC+FAK inhibitor(Y15) cream(100 mg+10 mg per rat) group, with 10 rats in each group. After the rat tail skin injury model was successfully constructed, in the treatment group, corresponding drugs were applied to the wound surface, while in the control and model groups, the same amount of cream base as the TGCC group was applied by the same method. Then, sterile gauze was wrapped around the wound edge, and these operations were performed three times a day for 28 consecutive days. The wound healing status at the third, seventh, eleventh, fourteenth, twenty-first, and twenty-eighth days was recorded, and the wound healing rate and healing time were calculated. On the day after the last dose of medication, rat serum and tail skin wound tissue were collected for analyzing the activities of serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), creatinine(CREA), urea, reactive oxygen species(ROS), interferon gamma(IFN-γ), interleukin(IL)-1β, IL-6, IL-4, IL-10, tumor necrosis factor(TNF)-α, as well as catalase(CAT), glutathione(GSH), lactate dehydrogenase(LDH), malondialdehyde(MDA), myeloperoxidase(MPO), superoxide dismutase(SOD), total antioxidant capacity(T-AOC), platelet endothelial cell adhesion molecule-1(CD31), and leukocyte differentiation antigen 34(CD34) in the wound tissue of rat tail skin. Hematoxylin-eosin, Masson, and sirius red staining were used to observe the morphological changes in the wound tissue of rat tail skin. The thickness of the epidermis, the number of fibroblasts and blood vessels, and the contents of collagen fibers, typeⅠ collagen(COLⅠ), and COLⅢ were calculated. The mRNA expressions of keratin 10(KRT10), KRT14, vascular endothelial growth factor(VEGF), fibroblast growth factor(FGF), epidermal growth factor(EGF), CD31, CD34, matrix metallopeptidase-2(MMP-2), MMP-9, COLⅠ, COLⅢ, desmin, fibroblast specific protein 1(FSP1), IFN-γ, IL-1β, TNF-α, IL-4, IL-6, and IL-10 in skin wound tissue were determined by quantitative real-time polymerase chain reaction(PCR). Western blot was utilized to detect the protein expressions of KRT10, KRT14, VEGF, FGF, EGF, MMP-2, MMP-9, COLⅠ, COLⅢ, desmin, FSP1, focal adhesion kinase(FAK), phosphorylated focal adhesion kinase(p-FAK), phosphatidylin-ositol-3-kinase(PI3K), phosphorylated phosphatidylin-ositol-3-kinase(p-PI3K), protein kinase B(Akt), phosphorylated protein kinase B(p-Akt), mammalian target of rapamycin(mTOR), and phosphorylated mammalian target of rapamycin(p-mTOR). The results manifest that TGCC can dramatically elevate the healing rate of rat tail wounds and shorten wound healing time. Besides, it can reduce serum ROS levels, the contents of MDA, MPO, and LDH in the rat skin wound tissue, as well as the serum IFN-γ, IL-1β, IL-6, and TNF-α levels and the mRNA expression levels of IFN-γ, IL-1β, IL-6, and TNF-α in the skin wound tissue. It can elevate the activities of CAT, GSH, SOD, and T-AOC in wound tissue, the IL-4 and IL-10 contents in serum, and the mRNA expressions of IL-4 and IL-10 in the wound tissue. In addition, TGGC can inhibit inflammatory cell infiltration and increase the epidermal thickness, counts of fibroblasts and blood vessels, and contents of collagen fibers, COLⅠ, and COLⅢ. Besides, TGCC can elevate the mRNA and protein expressions of epidermal differentiation markers(KRT10 and KRT14), endothelial cell markers(CD31 and CD34), angiogenesis and fibroblast proliferation, differentiation markers(VEGF, FGF, EGF, COLⅠ, COLⅢ, desmin, and FSP1), reduce the mRNA and protein expressions of gelatinases(MMP-2 and MMP-9), and increase protein expressions of p-FAK, p-PI3K, p-Akt, p-mTOR, as well as ratios of p-FAK/FAK, p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR. These results suggest that TGCC can significantly facilitate skin wound healing, and its mechanism may be related to the activation of the FAK/PI3K/Akt/mTOR signaling pathway, inhibition of inflammatory cell infiltration in skin wound tissue, elevation of epidermal thickness, counts of fibroblasts and vessels, and contents of collagen fiber, COLⅠ, and COLⅢ, and reduction of MMP-2 and MMP-9 expressions, thus accelerating wound healing.
Animals ; Male ; Wound Healing/drug effects* ; Rats ; Rats, Sprague-Dawley ; Signal Transduction/drug effects* ; TOR Serine-Threonine Kinases/genetics* ; Phosphatidylinositol 3-Kinases/genetics* ; Skin/metabolism* ; Proto-Oncogene Proteins c-akt/genetics* ; Tannins/pharmacology* ; Humans ; Drugs, Chinese Herbal/administration & dosage* ; Focal Adhesion Kinase 1/genetics*

As the brain's resident immune cells, microglia perform crucial functions such as phagocytosis, neuronal network maintenance, and injury restoration by adopting various phenotypes. Dynamic imaging of these phenotypes is essential for accessing brain diseases and therapeutic responses. Although numerous probes are available for imaging pro-inflammatory microglia, no PET tracers have been developed specifically to visualize anti-inflammatory microglia. In this study, we present an ¹⁸F-labeled PET tracer (QTFT) that targets the P2Y₁₂, a receptor highly expressed on anti-inflammatory microglia. [¹⁸F]QTFT exhibited high binding affinity to the P2Y₁₂ (14.43 nmol/L) and superior blood-brain barrier permeability compared to other candidates. Micro-PET imaging in IL-4-induced neuroinflammation models showed higher [¹⁸F]QTFT uptake in lesions compared to the contralateral normal brain tissues. Importantly, this specific uptake could be blocked by QTFT or a P2Y₁₂ antagonist. Furthermore, [¹⁸F]QTFT visualized brain lesions in mouse models of epilepsy, glioma, and aging by targeting the aberrantly expressed P2Y₁₂ in anti-inflammatory microglia. In a pilot clinical study, [¹⁸F]QTFT successfully located epileptic foci, showing enhanced radioactive signals in a patient with epilepsy. Collectively, these studies suggest that [¹⁸F]QTFT could serve as a valuable diagnostic tool for imaging various brain disorders by targeting P2Y₁₂ overexpressed in anti-inflammatory microglia.

AIM: To explore the effect of Huatanjiangqi capsule medicated serum (HTJQ) on the resistance of human bronchial epithelial cells (16HBE) to glucocorticoid (GC) stimulated by cigarette smoke extract (CSE). METHODS: After 16HBE cells were treated with HTJQ, the effects of different concentrations of HTJQ on the viability of 16HBE cells were determined by CCK-8 method. 16HBE cells were pretreated with HTJQ, and then cultured with dexamethasone (DEX) and lipopolysaccharide (LPS) for 24 hours, the effect of HTJQ on glucocorticoid (GC) resistance of 16HBE cells was determined by Enzyme-linked immunosorbent assay (ELISA). The effects of HTJQ, sulforaphane (SFN) and glutathione (GSH) on the expression of NF-E2-related factors 2 (Nrf2), Heme oxygenase-1 (HO-1) and histone deacetylase 2 (HDAC2) in 16HBE cells stimulated by CSE were measured by Western blot, and the effects of HTJQ, SFN and GSH on interleukin-8 (IL-8) in 16HBE cells were measured by ELISA. RESULTS: HTJQ promoted the proliferation of 16HBE cells at 1 h, 2 h and 4 h, the results of ELISA and Western blot showed that CSE induced GC resistance and decreased the expression of Nrf2, HO-1 and HDAC2 in 16HBE cells, HTJQ significantly decreased IL-8 and improved GC sensitivity of 16HBE cells (P<0.01), and up-regulated the expression of Nrf2, HO-1 and HDAC2 (P<0.01). In addition, HTJQ significantly up-regulated the level of GSH in 16HBE cells (P<0.01). Nrf2 agonists SFN and GSH significantly improved the glucocorticoid sensitivity of 16HBE cells (P<0.01), and up-regulated the expression of Nrf2, HO-1 and HDAC2 (P<0.01). CONCLUSION: HTJQ improves the GC resistance of 16HBE cells by up-regulating the expression of Nrf2/HDAC2 protein and the level of intracellular GSH.

Objective To investigate the secretory capacity and apoptosis of interleukin ( IL)-21 induced normal B cells by co-culture with serum from patients with systemic lupus erythematosus (SLE). Methods Serum from twenty new-onset SLE patients and 20 healthy donors were collected .CD1+9 B cells from the normal controls were co-cultured with serum from SLE patients in the presence or absence of IL-21-R-FC(4 μg/ml).Supernatant IgG and IgM concentration were measured by immunoturbidimetric assay on day 5.Supernatant anti-dsDNA level was determined by ELISA .The percentage of apoptotic cells was detected by flow cytometer.Results IgG,IgM and anti-dsDNA levels in normal B cells with SLE serum were significantly higher than those in the serum of SLE patients alone [ ( 5.84 ±1.79 ) g/L vs ( 4.25 ± 1.48)g/L,P=0.000;(0.46 ±0.21)g/L vs (0.43 ±0.21)g/L,P=0.003;(127.76 ±70.24)IU/ml vs (115.15 ±63.88) IU/ml,P=0.014 respectively].However, no significant differences were found in the group of normal B cells with non-homologous serum from normal controls (P>0.05).Supernatant IgG, IgM and anti-dsDNA levels in normal B cells with SLE serum significantly decreased while IL-21R-fusion protein was added [(5.26 ±1.62)g/L vs (5.84 ±1.79)g/L, P=0.006;(0.42 ±0.20)g/L vs (0.46 ±0.21)g/L, P=0.002;( 118.00 ±69.62 ) IU/ml vs ( 127.76 ±70.24 ) IU/ml, P =0.012 respectively ] .The apoptotic rate of B cells with SLE serum was significantly higher than that with normal serum [ ( 47.88 ± 12.65)%vs (38.86 ±10.32)%,P =0.004].But adding IL-21R-fusion conversed the apoptotic rates [(42.08 ±12.52)%vs (47.88 ±12.65)%,P=0.001].Conclusions SLE serum could induce normal B cells to form immunoglobulin secreting cells and producing autoantibodies , or apoptosis in pathological conditions.IL-21 might be considered as a potential therapeutic target of SLE .

Objective:To evaluate the efficacy and safety of cytoreductive surgery (CRS) in conjunction with hyperthermic intra-peritoneal chemotherapy (HIPEC) for treating patients with peritoneal carcinomatosis (PC) from colorectal cancer (CRC). Methods:A total of 62 CRC patients with complication of PC were divided into the CRS group, namely, Group One (n=29, CRS and systemic adju-vant chemotherapy) and the CRS+HIPEC group, namely, Group Two (n=33, CRS+HIPEC). The primary end point of the study was overall survival (OS) and the secondary end point was serious adverse events (SAE). Results:Patients' clinicopathologic characteris-tics, peritoneal carcinomatosis index, and completeness of cytoreduction therapy were well balanced and comparable between the two groups. The median follow-up was 41.9 mo (6.5 mo to 110.0 mo) in Group One and 32.0 mo (10.5 mo to 95.9 mo) in Group Two. The median OS was 8.5 mo (95%CI:4.9 mo to 12.1 mo) in Group One and 14.5 mo (95%CI:11.9 mo to 17.1 mo) in Group Two (P=0.007). Within 30 days after the surgery, SAE occurred in 3 of the 29 patients in Group One, and 9 of the 33 patients in Group Two (P=0.126). Multivariate analysis revealed that HIPEC, CC0-1 score, and chemotherapy over six cycles were the independent factors for OS improvement. Conclusion:The CRS+HIPEC method improves the OS of patients with PC from CRC, suggesting an acceptable safety.

This study is to investigate the effect of the major chemical composition in rhizome of Pterocypsela elata, lactuside B, on expression of bcl-2, bax mRNA and their protein in rats' cerebral cortex after cerebral ischemia-reperfusion injury. First, middle cerebral artery ischemia-reperfusion injury model was established, and each group was treated with the corresponding medicines. Animals were separately sacrificed at 24 h and 72 h. The brain infarct volumes were detected by TTC dye, bcl-2 and bax mRNA expression was checked by RT-PCR, and the proteins of bcl-2 and bax were explored by two-step immunohistochemistry in cerebral cortex of rats. Lactuside B can reduce brain infarct volume of cerebral cortex of rats, increase the expression of bcl-2 mRNA and decrease that of bax mRNA. Moreover, the ratio of bcl-2 to bax mRNA is higher in 12.5 and 25 mg kg(-1) dose group, respectively, which is significantly different from that of model group (P < 0.05 or P < 0.01). Generally, either 12.5 or 25 mg kg(-1) dose group is better than positive control medicine nimodipine (P < 0.05 or P < 0.01). In addition, the expression of bcl-2 and bax protein is consistent with their gene expression. Infarct volume and the ratio of bcl-2 to bax mRNA expression are significantly different (P < 0.05 or P < 0.01) between 72 h and 24 h group. The results demonstrated that lactuside B could play a good role in resisting cerebral ischemia by upregulating the expression of bcl-2 mRNA and protein and downregulating that of bax mRNA and protein.

Objective To study the clinical, laboratory, radiological and pathological findings of patients with hypertrophic cranial pachymeningitis (HCP) in Wegner's granulomatosis (WG) to improve the recognition of the disease, even when it occurs in limited form. Methods Three patients were described and English literatures of biopsy-proven pachymeningitis in WG were reviewed. Results The features of WG-associated pachymeningitis included: ① Frequently occurred early in the course of active limited WG; ② Commonly presented with sever headache and cranial neuropathies in the absence of other meningeal irritative signs; ③ Variable cerebrospinal fluid findings with mild predominantly lymphocytic pleocytosis and elevated protein concentration were major laboratorg findings; ④Elevated ESR and positive serum anti-neutrophilic cytoplasmic antibody (ANCA) could be found in most patients; ⑤ Gadolinium-enhanced brain MRI is very senitive in the detection of pachymeningitis; ⑥A dural biopsy showed granulomatous necrotizing inflammation, giant cell, and evidence of vasculitis;⑦ A favorable response to standard treatment with corticosteroid, cyclophosphamide or other cytotoxic drugs could be observed. Conclusion HCP may be the initial or cardinal manifestation of the limited form of WG. Early diagnosis by ANCA, MRI and dural biopsy may facilitate diagnosis Corticosteroid and immunosupressant are the choices of treatment.

Objective To study the changes of clonality of T cell receptor (TCR) and complementarity determinative region 3(CDR3) before and after autologous peripheral blood CD34+ stem cell transplantation(auto-PBSCT) for severe/refractory connective tissue disease(CTD). Methods Thirteen patients with severe/refractory CTD were enrolled for auto-PBSCT in Peking Union Medical College Hospital, including systemic lupus erythematosus (8 cases), rheumatoid arthritis(4 cases), and primary Sjogren’s Syndrome(1 case). Blood samples were collected before/after mobilization, 2 weeks, 1, 3, 6, 12 and 18 months post-transplantation. Diversity of TCRBV and CDR3 were showed by reverse transcription-polymerase chain reaction(RT-PCR) and genescan. Results The TCR BV usage and CDR3 spectral pattern of pre-auto-PBSCT CTD patients were revealed skewed pattern and oligoclonality, Which developed severe oligoclonality within 1 months after auto-PBSCT. However, they showed diversity andpoly-clonality 3～6 months after auto-PBSCT. Conclusion Skewed pattern and oligoclonality of TCRBV and CDR3 which implied auto-reactive were depressed after auto-PBSCT, and inclined to change to normal pattern.

Objective To investigate the prevalence,their autoantigen and the clinical significants of antiendothelial cell antibodies(AECAs) in systemic vasculitis.Methods Western blotting was performed to detect specific AECA in serum of systemic vasculitis,SLE,RA,SS and healthy donors.Then to analyze the relationships of AECA with the disease manifestation.Results(1)The prevalence of AECA was 77.7% in systemic vasculitis,87.5% in SLE,66.7% in SS,7.14% in RA and 10% in normal group respectively.(2)AECA reacted with a heterogeneous series of endothelial proteins which ranged in molecular size from 16 to 120 ku Furthermore,AECA against a 47 ku endothelial cell antigen were more frequently found in a variety of systemic vasculitis and SLE.(3)Compared with those in AECA-negative patients,the mean levels of ESR in AECA-positive patients with TA and the mean levels of BVAS in patients with WG,MPA and CSS were both significantly higher in AECA-positive patients.Patients with BD who have AECA against 47 ku endothelial cell proteins were more frequently found to have neuropathy than those 47 ku-AECA-negative patients,and the prevalence of inhanced CRP are also more frequent.Conclusion AECA showed to be correlated with the disease manifestation,and the same molecular sizeantigen could be found in a variety of systemic vasculitis and SLE.

Objective To investigate the clinical significance of antiendothelial cell antibodies (AECA) in systemic vasculitis. Method With Human umbilical vein endothelial cell (HUVEC) as substrate cell, sera from 129 systemic vasculitis patients [including 59 Behcet′s disease(BD), 28 Takayasu arteritis (TA), 20 Wegener′s granulomatosis (WG), 8 polyarteritis nodosa (PAN), 9 microscopic polyangiitis (MPA), 5 Churg-Strauss syndrome (CSS)] were screened for the presence of AECA by ELISA. Sera from SLE, RA and healthy donors were examined as controls. The association of AECA to clinical disease activity was analyzed. Result The prevalence of AECA by HUVEC cell-ELISA was 59% in systemic vasculitis [48% in BD,79% in TA, 65% in WG, 63% in PAN, 44% in MPA, 80% in CSS], 46% in SLE, 4% in RA, and 2.4% in control group. Compared with patients with RA and control group, AECA were more frequently found in patients with systemic vasculitis and SLE (P