OBJECTIVEIn order to explore the roles of Huoxueruanjian compound on liver fibrogenesis and its molecular mechanism, this paper has investigated the Influence of blood serum with such traditional Chinese medicine compound on the expression of Smad3, Smad7 and procollagen alpha2(I) gene in hepatic stellate cell (HSC).

METHODSHSC-T6 was deal with different Concentration of blood serum medicine with Heluoshugan which was made by routine way. Then expression change of Smad3, Smad7 and procollagen alpha2(I) mRNA among each groups were observed by RT-PCR. Furthermore, the expression change of Smad3 protein were examined by Western blot.

RESULTSExpression of Smad3 and procollagen alpha2(I) mRNA as well as Smad3 protein had been downregulated after treating with blood serum medicine of Heluoshugan (P<0.01, P<0.05, respectively). The expression of procollagen alpha2(I) mRNA changed at the same tendency as those of Smad3. The role of blood serum medicine was significant difference between different concentration, P<0.05. And the expression of procollagen alpha2(I) mRNA changed in concentration-dependent manner. Blood serum medicine has no effects on the Smad7 mRNA.

CONCLUSIONThe anti-fibrosis roles of HuoXueruanjian Compound maybe influence the function of TGF-beta and Smad by nonspecific action, thereby inhibit the transcription of procollegan alpha2(I) mRNA and decrease the production of ECM. As regards Smad3, it may be facilitating the development of liver fibrosis when its expression increases. Otherwise, it manifest with anti-fibrosis role.

Animals ; Collagen Type I ; genetics ; DNA-Binding Proteins ; genetics ; Liver Cirrhosis ; drug therapy ; etiology ; pathology ; Medicine, Chinese Traditional ; RNA, Messenger ; analysis ; Rats ; Smad3 Protein ; Smad7 Protein ; Trans-Activators ; genetics

BACKGROUND/AIMS: Previous studies have revealed that mast cells (MCs) may activate the protease-activated receptors and release of neuropeptides involved in the pathogenesis of irritable bowel syndrome (IBS). The levels of protease-activated receptor 2 (PAR-2) and tryptase can contribute to understanding the pathogenesis of IBS. METHODS: Colonoscopic biopsies were performed of 38 subjects (20 with IBS-diarrhea [IBS-D], eight with IBS-constipation [IBS-C], and 10 healthy volunteers). The mRNA and protein levels of tryptase and PAR-2 were assessed by real-time PCR and Western blot. The levels of vasoactive intestinal peptide (VIP), substance P (SP), and calcitonin gene-related peptide (CGRP) were measured by immunohistochemistry, and MCs were counted by toluidine blue staining. RESULTS: Significant increases in the mRNA expression of tryptase (p<0.05, IBS-D, IBS-C vs control) and PAR-2 (p<0.05, IBS-D, IBS-C vs control) and in the tryptase protein level (p<0.05, IBS-D, IBS-C vs control) were detected in IBS. Elevations of MCs, CGRP, VIP and SP (p<0.05, IBS-D vs control) were observed for IBS-D only. CONCLUSIONS: Tryptase levels may upregulate the function of PAR-2, resulting in the release of neuropeptide and they were correlated with clinical symptoms associated with IBS.
Biopsy ; Blotting, Western ; Calcitonin Gene-Related Peptide ; Immunohistochemistry ; Inflammation ; Irritable Bowel Syndrome* ; Mast Cells ; Neuropeptides ; Real-Time Polymerase Chain Reaction ; Receptor, PAR-2* ; Receptors, Proteinase-Activated ; RNA, Messenger ; Substance P ; Tolonium Chloride ; Tryptases* ; Vasoactive Intestinal Peptide

OBJECTIVETo evaluate the clinical outcomes between extreme lateral interbody fusion and conventional posterior operation in the treatment of upper lumbar disc herniation.

METHODSAmong 60 patients with upper lumbar disc herniation were treated with extreme lateral interbody fusion(XLIF) or conventional posterior operation from June 2010 to December 2014, 30 patients(19 males and 11 females) were treated with XLIF (XLIF group); and the other 30 patients(17 males and 13 females) were treated with conventional posterior operation (conventional group). In XLIF group, the lesions occurred at T₁₂L₁ segments in 2 patients, at L₁,₂ segments in 6 patients, at L₂,₃ segments in 10 patients, and at L₃,₄ segment in 12 patients. In conventional group, the lesions occurred at T₁₂L₁ segments in 1 patient, at L₁,₂ segments in 6 patients, at L₂,₃ segments in 8 patients, and at L₃,₄ segment in 15 patients. Operative incision lengths, time, blood loss, postoperative draining volume, hospital stays were recorded. Pre-and post-operative visual analogue score(VAS) and Japanese Orthopedic Association(JOA) were compared between two groups. According to the image data, the intervertebral fusion device was observed to be displaced and the rate of interbody fusion was analyzed.

RESULTSAll the patients were followed up, and the duration ranged from 12 to 48 months, with an average of 29 months. The complications included 2 femoral nerve damage in XLIF group (postoperative recovery within 3 months) and superficial incision infection in conventional group(cured by anti-infection). There were no patients with cerebrospinal fluid leakage(CSFL), cauda equina injuries or functional deterioration in the nerve root of lower limbs. In the XLIF group: the operative time was (65.6±20.5) minutes, blood loss was (48.8±15.3) ml, postoperative draining volume was 0 ml. In the conventional group: the operative time was (135.2±33.9) minutes, blood loss was (260.3±125.7) ml, postoperative draining volume was (207.1±50.2) ml. The operative time, blood loss, postoperative draining volume in XLIF group were less than those in the conventional group(<0.05). The JOA and VAS score were significantly improved in both groups during the follow-up period compared with those before operation(<0.05). But the difference of the JOA and VAS score between the two groups 1, 6, and 24 months after surgery had not significant differences(>0.05). There were no significant differences in the fusion rate between the two groups 6 and 12 months after operation(>0.05).

CONCLUSIONSThe XLIF fusion for the treatment of upper lumbar disc herniation has several advantages such as minimal invasive, stable vertebral plate, less complications and postoperative fusion rate, which has a better clinical effect.

OBJECTIVETo detect the expression of membrane-bound HLA-G (mHLA-G) and serum HLA-G (sHLA-G) in acute leukemia patients and investigate the correlation between HLA-G expression and the occurrence and development of acute leukemia.

METHODSEnzyme-linked immunosorbent assay and flow cytometry were used to detect the expression levels of sHLA-G and mHLA-G in 40 newly diagnosed leukemia cases, 10 refractory and relapsed leukemia cases, and 30 leukemia cases receiving chemotherapy. Ten normal individuals served as the normal control group.

RESULTSThe mean serum level of sHLA-G in normal individuals was 5.87±2.07 ng/ml, as compared to 10.05±6.58 ng/ml in newly diagnosed leukemia patients and 12.32±5.85 ng/ml in refractory and relapsed cases. The mean level of mHLA-G in normal individuals, newly diagnosed cases, and refractory and relapsed cases were (0.29±0.20)%, (0.60±0.44)%, and (0.77±0.41)%, respectively. The mean levels of sHLA-G and mHLA-G were significantly higher in the newly diagnosed cases than that in the normal controls (P<0.05), and significantly higher in patients before chemotherapy than in those with complete remission after chemotherapy (P<0.05).

CONCLUSIONHLA-G expression levels might influence the treatment outcomes and can serve as a prognostic factor for acute leukemia.

Acute Disease ; Adolescent ; Adult ; Aged ; Case-Control Studies ; Female ; HLA-G Antigens ; blood ; metabolism ; Humans ; Leukemia ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; Young Adult

OBJECTIVETo investigate the effect of granulocyte colony stimulating factor (G-CSF) on myeloid-derived suppressor cells (MDSCs) in the bone marrow and peripheral blood, and explore the relationship between MDSC and graft-versus-host disease (GVHD).

METHODSBone marrow, peripheral blood and peripheral blood stem cells were obtained from 12 healthy hemopoietic stem cell donors before and on day 5 after G-CSF mobilization. Flow cytometry was employed to examine the number of MDSC, and the relationship between MDSC number and the incidence of GVHD was analyzed.

RESULTSIn normal physiological conditions, MDSC could be detected in the peripheral blood and bone marrow with a cell percentages of (1.35±0.35)% and (2.44±1.11)%, respectively, showing a significantly higher cell percentage in the bone marrow (P=0.015). On the 5th day after G-CSF mobilization, the percentage of MDSCs increased to (4.01±1.82)% in the peripheral blood and to (4.38±2.19)% in the bone marrow, showing no significant difference between them (P=0.083). The mobilization caused a significant increase in the number of MDSCs in the peripheral blood (P=0.047) but not in the bone marrow (P=0.761). The number of MDSCs in the collected samples showed a significant inverse correlation to the incidence of GVHD (P=0.048).

CONCLUSIONSMDSCs are present in the peripheral blood and bone marrow of healthy donors, with a greater number in the bone marrow. G-CSF can mobilize the MDSCs from the bone marrow to the peripheral blood to increase number of MDSCs in the peripheral blood, which may contribute to a lowered incidence of GVHD in hematopoietic stem cell transplantation (HSCT).

Adolescent ; Adult ; Bone Marrow Cells ; cytology ; Female ; Graft vs Host Disease ; prevention & control ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; T-Lymphocytes ; cytology ; drug effects ; metabolism ; Young Adult

OBJECTIVETo study the optimal surgical resection length for esophageal carcinoma.

METHODSSpecimens of seventy patients with esophageal squamous cell carcinoma resected and collected in our hospital were made into pathologic giant sections. Direct intramural infiltration, multicentric carcinogenic lesion and leaping metastasis were observed in the large slice by microscope. The actual length during the operation was calculated by the ratio of shrinkage.

RESULTSDirect intramural infiltration was found in 51 (72.9%) patients, 39 proximal and 36 distal to the tumor. The mean length of direct intramural infiltration was 0.9 +/- 0.8 cm (4.0 cm maximum) proximally and 0.5 +/- 0.3 cm (2.0 cm maximum) distally. Multicentric carcinogenic lesion was found in 11 (15.7%) patients, 5 proximally, 8 distally and 2 on both sides. Proximal to the tumor, the mean distance between the multicentric carcinogenic lesion and the main lesion plus the length of the multiple carcinogenic lesion was 3.2 +/- 1.5 cm (4.7 cm maximum). Distal to the tumor, it was 3.6 +/- 2.4 cm (9.1 cm maximum). Leaping metastasis was found in 9 (12.9%) patients, 7 proximally and 4 distally. The mean distance between the leaping metastasis and the main lesion plus the length of the leaping metastatic lesion was 1.9 +/- 0.6 cm (2.9 cm maximum) proximally and 1.4 +/- 1.0 cm (2.7 cm in maximum) distally.

CONCLUSIONThe optimal surgical resection length for esophageal carcinoma should be at least 5 cm proximal to the tumor and total length on the distal side.

Esophageal Neoplasms ; pathology ; surgery ; Female ; Humans ; Male ; Neoplasm Invasiveness

To investigate the protective effect of spermine (Sp) on diabetic cardiomyopathy (DCM) and high glucose-induced cardiac fibroblasts (CFs), and to explore its mechanism. ①Animal experiments: 24 male Wistar rats were randomly divided into control group, type 1 diabetes group (TID) and spermine group (TID+Sp, each group n=8). TID rats were induced by streptozocin (STZ, 60 mg/kg), and TID+Sp rat were pretreated with spermine (Sp, 5 mg/(kg·d)) for 2 weeks before STZ injection. After 12 weeks of modeling, blood glucose, insulin levels, ejection fraction (EF) and shortening fraction (FS) were measured, and Masson staining and Sirius red staining were performed in the rat cardiac tissues. ②Cell experiments: primary CFs were extracted from newborn (1-3 d) Wistar rat hearts, and were randomly divided into control group, high-glucose group (HG) and HG+Sp group (n=6 per group). HG group was treated with 40 mmol/L glucose, and the HG+Sp group was pretreated with 5 μmol/L Sp for 30 min before HG treatment. The cell viability of CFs was detected by CCK8, the content of collagen in culture medium was analyzed by ELISA, and protein expressions of cell cycle related proteins (PCNA, CyclinD1 and P27) were detected by Western blot. Compared with control group, the blood glucose and collagen content were increased, and the insulin level and heart function were decreased in the T1D group. Meanwhile, HG induced an increasing of the cell viability, the collagen content in the medium and the expressions of PCNA and CyclinD1, while the expression of P27 was down-regulated. Spermine could reduce the above changes, manifested as improving the cardiac function, regulating the expression of cyclin and reducing the level of myocardial fibrosis. Spermine can alleviate myocardial fibrosis in diabetic cardiomyopathy, which mechanism is related to the regulation of cell cycle.

It is investigated that the hepatotoxicity of Polygonum multiflorum (PM)was attenuated by its processed products of nine times steaming and nine times sunning(RPM)based on immunological stress-mediated animal model by using metabolomics method. Sprague-Dawley(SD)rats were intragastrically administered with(5.4 g crude drug per kg body weight)of 50% alcohol extracts of PM and its processed products of nine times steaming and nine times sunning respectively or co-treated with non-toxic dose of lipopolysaccharide(LPS, 2.8 mg·kg^-1)via tail vein injection. The plasma alanine aminotransferase(ALT)and aspartate aminotransferase(AST)activities were assayed and the isolated livers were evaluated for histopathological changes. Global metabolomics profiling, multivariate analysis and data base searching were performed to discover common differential metabolites for idiosyncratic liver injury. The results showed that co-treatment with non-toxic dose of LPS and PM could result in significant liver injury, indicated by the elevation of plasma ALT and AST activities, as well as obvious liver histologic damage; whereas RPM failed to induce detectable liver injury. Furthermore, 10 potential metabolomics biomarkers that differentially expressed in LPS/PM group compared with LPS/RPM without liver injury were identified by untargeted metabolomics, mainly involved ten pathways: sphingolipid metabolism, linoleic acid metabolism, taurine and hypotaurine metabolism, steroid hormone biosynthesis, galactose metabolism, steroid biosynthesis, metabolism of xenobiotics by cytochrome P450, pyrimidine metabolism, biosynthesis of unsaturated fatty acids, primary bile acid biosynthesis. This work illustrated the idiosyncratic hepatotoxicity of heshouwu and provided a metabolomic insight into diosyncratic liver injury of PM and RPM.

Cyclophosphamide (CPA) is one of the most commonly used alkylating agents in the treatment of malignant cancer. CPA is metabolized by cytochrome P450 enzymes into 4-hydroxycyclophosphamide in vivo which can exhibit anti-tumor activity. Metabolic activation of CPA can cause adverse reactions such as myelosuppression, cystitis, and liver injury. The aim of this study was to evaluate the dynamic changes of hepatic injury induced by CPA in mice. Male BALB/c mice were injected CPA (200 mg·kg^-1) intraperitoneally. Both serum and liver samples were collected at 0, 2, 6, 12 and 24 hours after dosing. The animal experiment protocol was approved by the Institutional Animal Care and Use Committee at Sun Yat-sen University. The results showed that hepatotoxicity was observed at 2 hours after CPA dosing, and the most serious liver injury was measured at 12 hours. The level of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and malondialdehyde (MDA) was significantly increased, glutathione (GSH) level was significantly decreased, hepatocyte edema and vacuolar degeneration were widely observed in liver tissue, and began to recover 24 hours after dosing. In addition, due to oxidative stress damage caused by CPA, nuclear factor-erythroid 2-related factor 2 (NRF2) signaling pathway was activated and the mRNA and protein expression of its downstream targets such as quinine oxidoreductase 1 (NQO1), heme oxygenase-1 (HO-1), glutamate-cysteine ligase catalytic subunit (GCLC) and glutamate cysteine modifier subunit (GCLM) were up-regulated, which alleviated oxidative stress injury. In a summary, this study demonstrate the dynamic change of CPA-induced liver injury and the NRF2-mediated protective mechanisms, providing new insights into the CPA-induced liver injury.

Objective: To investigate the functional outcomes and postoperative complications of Cheng's GIRAFFE reconstruction after proximal gastrectomy. Methods: A descriptive case series study was conducted. Clinical data of 100 patients with adenocarcinoma of the esophagogastric junction who underwent Cheng's GIRAFFE reconstruction after proximal gastrectomy in Cancer Hospital of University of Chinese Academy of Sciences (64 cases), Zhejiang Provincial Hospital of Chinese Medicine (24 cases), Lishui Central Hospital (10 cases), Huzhou Central Hospital (1 case) and Ningbo Lihuili Hospital (1 case) from September 2017 to June 2021 were retrospectively analyzed. Of 100 patients, 64 were males and 36 were females; the mean age was (61.3 ± 11.1) years and the BMI was (22.7±11.1) kg/m(2). For TNM stage, 68 patients were stage IA, 24 were stage IIA and 8 were stage IIB. Postoperative functional results and postoperative complications of radical gastrectomy with Giraffe reconstruction were analyzed and summarized. Gastroesophageal reflux disease questionnaire (RDQ) score and postoperative endoscopy were used to evaluate the occurrence of reflux esophagitis and its grade (grade N, grade A, grade B, grade C, and grade D from mild to severe reflux). The continuous data conforming to normal distribution were expressed as (mean ± standard deviation), and those with skewed distribution were presented as median (Q1, Q3). Results: All the 100 patients successfully completed R0 resection, including 77 patients undergoing laparoscopic surgery and 23 patients undergoing laparotomy. The Giraffe anastomosis time was (38.6±14.0) min; the blood loss was (73.0±18.4) ml; the postoperative hospital stay was 9.5 (8.2, 13.0) d; the hospitalization cost was (6.0±0.3) ten thousand yuan. Fourteen cases developed perioperative complications (14.0%), including 7 cases of pleural effusion or pneumonia, 3 cases of anastomotic leakage, 2 cases of gastric emptying disorder, 1 case of gastrointestinal hemorrhage and 1 case of anastomotic stenosis, who were all improved and discharged after symptomatic management. Patients were followed up for (33.3±1.6) months. Eight patients were found to have reflux symptoms by RDQ scale six months after surgery, and 11 patients (11/100,11.0%) were found to have reflux esophagitis by gastroscopy, including 6 in grade A, 3 in grade B, and 2 in grade C. All the patients could control their reflux symptoms with behavioral guidance or oral PPIs. Conclusion: Cheng's GIRAFFE reconstruction has good anti-reflux efficacy and gastric emptying function; it can be one of the choices of reconstruction methods after proximal gastrectomy.
Adenocarcinoma/surgery* ; Aged ; Esophageal Neoplasms/surgery* ; Esophagitis, Peptic/etiology* ; Esophagogastric Junction/surgery* ; Female ; Gastrectomy/methods* ; Gastroesophageal Reflux/etiology* ; Humans ; Laparoscopy ; Male ; Middle Aged ; Reconstructive Surgical Procedures/methods* ; Recovery of Function ; Retrospective Studies ; Stomach Neoplasms/surgery*