A new benzene derivative microintegerrin C (1) and a new norsesquiterpenoid microintegerrin D (2), along with six known compounds (3-8), were isolated and identified from stems and leaves of Micromelum integerrimum by various chromatographies such as silica gel, Sephadex LH-20, RP-18 column chromatography and HPLC. Their structures were mainly identified based on the spectral data analysis such as 1D-, 2D-NMR and HR-EI-MS. All known compounds were isolated from this plant for the first time.
Chromatography, High Pressure Liquid ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Rutaceae ; chemistry ; Sesquiterpenes ; isolation & purification

To study the hemolytic effect of polyphyllin II (PP II) mediated by anion channel protein and glucose transporter 1 (GLUT1), in order to initially reveal its hemolytic mechanism in vitro. In the experiment, the spectrophotometric method was adopted to detect the hemolysis of PP II in vitro and the effect of anion channel-related solution and blocker, glucose channel-related inhibitor and multi-target drugs dehydroepiandrosterone (DHEA) and diazepam on the hemolysis of PP II. The scanning electron microscope and transmission electron microscope were used to observe the effect of PP II on erythrocyte (RBC) morphology. The results showed that PP II -processed blood cells were severely deformed into spherocytes, acanthocyturia and vesicae. According to the results of the PP II hemolysis experiment in vitro, the anion hypertonic solution LiCl, NaHCO3, Na2SO4 and PBS significantly inhibited the hemolysis induced by PP II (P < 0.05), while blockers NPPB and DIDS remarkably promoted it (P < 0.01). Hyperosmotic sodium chloride, fructose and glucose at specific concentrations notably antagonized the hemolysis induced by PP II (P < 0.05). The glucose channel inhibitor Cytochalasin B and verapamil remarkably antagonized the hemolysis induced by PP II (P < 0.01). The hemolysis induced by PP II could also be antagonized by 1 gmol x L(1) diazepam and 100 μmol x L(-1) DHEA pretreated for 1 min (P < 0.01). In conclusion, the hemolytic mechanism of PP II in vitro may be related to the increase in intracellular osmotic pressure and rupture of erythrocytes by changing the anion channel transport activity, with GLUT1 as the major competitive interaction site.
Animals ; Diosgenin ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Erythrocytes ; cytology ; drug effects ; Hemolysis ; drug effects ; Hemolytic Agents ; pharmacology ; Sheep

Objective To measure the renal tissue texture or flexibility with virtual touch quantization (VTQ) and to tentatively examine its clinical application in patients with chronic kidney disease(CKD).Methods A total of 750 patients (1500 kidneys) were performed with VTQ,including 400 cases in the control group,and 350 cases in the CKD group.A conventional ultrasound examination (two-dimensional,color Doppler) were first taken,and then the shear wave velocity (Vs) was measured which reflected the textural elastic.Results In both groups the Vs was the highest in renal cortex with significant difference (P<0.05); renal cortical region Vs in CKD group was lower than those in control group (P<0.05),while Vs of renal medulla and renal sinus had no significant difference in the two groups.The severity of renal dysfunction was increased along with a Vs decrease of renal cortex.Conclusion VTQ is helpful to assess renal function of patients with CKD.

Objective To analyze indications,complications and outcomes of amputation.Methods A total of 15 patients undergone amputation in field or at tent hospital were collected for analy-zing injury severity,place where amputation was done,whether open or closed amputation and stitch re-moval time. Results There were 9 males and 6 females.at an average age of 32 years(11-51years).There were 16 amputations including Gustilo IIIB in 2 patients, Gustilo IIIC in 9 and Tscheme Ⅲin 5 according to Gustiln classification or Tscheme classification.Four patients who received amputation in field or at tent hospital developed infection and had to receive amputation again at a higher level on the limb and drainage of open wounds because of a higher infection rate due to the amputation location.Ten patients received first amputation at higher levels with open wound at station hospital but only 2 manifested infected incision.High level amputation with one stage closure was done in 1 patient who was infected and suppurated after operation and even developed bacteremia. Conclusions Infection rate following am-putation 4n field and tent clinics is rather higher,so secondary open amputations should be performed at a higher level as soon as possible.One-time and high-level open amputation plays an important role in treat-ment of severe lower limb injuries following earthquake.

OBJECTIVETo evaluate the efficacy of minocycline as an adjunct to scaling and root planning (SRP) in treating chronic periodontitis.

METHODS64 male smokers with moderate to advanced periodontitis were randomly divided into two groups: SRP alone (SRP) and SRP plus minocycline (SRP+M). All clinical parameters including plaque index (Pll), gingival index (GI), bleeding on probing (BOP), probing depth (PD) and attachment gain were recorded at baseline, 3 and 6 months after treatment.

RESULTSAccording to PlI, GI and BOP, there was no significant difference between the two groups at 3 and 6 months after periodontal therapy (1 > 0.05). However, PD reduction and attachment gain were significantly greater for SRP+M than that for SRP (P < 0.05). For SRP+M and SRP groups, PD reduction were 1.98 mm and 1.32 mm, and attachment gain were 1.87 mm and 1.14 mm respectively. Deep pockets in SRP+M group showed more obvious PD reduction (3.48 mm versus 2.21 mm, P < 0.01) and attachment gain (2.62 mm versus 1.23 zmm, P < 0.01) than that in SRP group.

CONCLUSIONTreatment with SRP plus locally delivered minocycline is more effective than SRP alone in male smokers with chronic periodontitis. Mechanical debridement plus locally delivered antibiotics are recommended especially for smokers with deep pocket periodontitis.

Adult ; Anti-Bacterial Agents ; Chronic Periodontitis ; Dental Plaque Index ; Dental Scaling ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Minocycline ; Periodontal Attachment Loss ; Periodontal Index ; Periodontal Pocket ; Periodontitis ; Root Planing

OBJECTIVE Human metapneumovirus (hMPV) is semblable to respiratory syncytial virus (RSV) which causes respiratory infections typically characterized by cough, runny nose, fever, and nasal congestion but sometimes progressing to bronchiolitis and pneumonia. Whereas, there is no corresponding drug to inhabit the virus. Studies of new compounds with potential anti-HMPV activity could produce clinical value. Chinese herbal medicine played a great role during COVID-19, therefore we choose some small molecular (JH001) extracted from botany to investigate therapeutic effect on hMPV and the underlying mechanisms. METHODS In this study, 16HBE cells were used as a model to explore in vitro antiviral effect. Cytotoxicity assays were performed before the antiviral tests, cell viability of 16HBE cells handled by different concentration of JH001 was estimated by Cell Counting Kit-8 (CCK-8). Then RT-qPCR, immunofluores?cence, and flow cytometer were used to test the viral titer after cells infected with hMPV. Eventually, 6-8 weeks mice were infected intranasally with 60 μL of hMPV, the control group was treated with 0.9％ saline water, other groups were administered with JH001 and ribavirin, then the lung virus titer and protective effect in lung were judged. RESULTS The obtained JH001 exhibited no cytotoxicity to 16HBE cells during 6.25 - 200 μmol · L-1. RT-QPCR demonstrated that JH001 showed obvious inhabitation to the viral replication and showed great significance compared with saline. And fluo?rescence exhibited distinct decrease of hMPV-N protein, flow cytometer results showed that MFI decrease evidently. Sig?nificant reduction of N-gene expression was observed in those mice treated with JH001 compared with saline group, which indicated that JH001 probably had protective and therapeutic effect on viral replication. CONCLUSION This study illustrated that JH001 might be a promising option for small molecular against hMPV and JH001 might be worthy of fur?ther development and used as a potential therapeutic strategy for other respiratory viruses in the future.

OBJECTIVETo assess the prognostic factors associated with the sudden idiopathic sensorineural hearing loss, to predict the prognosis of patient with idiopathic sensorineural hearing loss precisely.

METHODSEight hundreds and eighty two patients with idiopathic sudden sensorineural hearing loss were retrospectively reviewed during January 2006 to March 2007. Patients whose initial hearing threshold < or =40 dB were excluded. The patients with initial hearing threshold >40 dB were recruited, which was divided into six subgroups based on the patterns of audiogram: downgrade audiogram subgroup, upgrade audiogram subgroup, flat audiogram subgroup, concave audiogram subgroup, profound audiogram subgroup and total deafness subgroup.

RESULTSRegarding to the relationship between the time point for initial intervention and the prognosis, better prognosis was obtained in patients whose initial intervention was within 3 days of the disease, good prognosis was achieved within 1 or 2 weeks of the disease, poor prognosis was noted beyond 2 weeks of this disease. Furthermore, comparison with the initial intervention within 3 week, 1 month and 1 month later, the prognosis among them was not statistical different. 97.7% hearing recovery was achieved in the concave subgroup with the initial hearing threshold >40 dB group. Comparison with the other subgroup (except total deafness subgroup), the cure rate and recovery rate was 23.8% and 57.9% respectively in the profound subgroup. Poor prognosis was demonstrated in the total deafness subgroup and inefficacy rate was 67.4% in the total deafness subgroup. Comparison with patients without companying complications, the prognosis of patients with companying complications such as diabetes or high blood pressure has negative impact in hearing recovery. The age was correlated with the prognosis, elder had poor prognosis, patients more than 50 years old present with worse hearing than that less than 50 years old (H = 7.851, P = 0.0051).

CONCLUSIONSThe initial intervention beyond 2 weeks had negative impact on the prognosis. The initial audiogram patterns and hearing threshold were both significant factors on the prognosis of idiopathic sudden sensorineural hearing loss. In addition, old patient had poor prognosis. The companying complications such as high blood pressure and diabetes had negative impact on the prognosis of idiopathic sudden hearing sensorineural loss.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Audiometry ; Auditory Threshold ; Child ; Female ; Hearing Loss, Sensorineural ; complications ; diagnosis ; physiopathology ; Hearing Loss, Sudden ; complications ; diagnosis ; physiopathology ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Risk Factors ; Young Adult

OBJECTIVETo observe the influence of high mobility group box-1 protein (HMGB1) derived from spleen on the phenotype of regulatory T lymphocytes (Treg) and HMGB1-mediated immune function in severely scalded rats after delayed resuscitation.

METHODSOne hundred and four Wistar rats were divided into normal control group (NC, n = 8), sham scald group (SS, n = 32), scald group (S, n = 32), and ethyl pyruvate (EP) treatment group (EPT, n = 32) according to the random comparison table. Rats in the latter 2 groups were subjected to 30%TBSA full-thickness scald, which were intraperitoneally injected with Ringer solution or EP solution at post scald hour (PSH) 6 (delayed antishock treatment) and administered with 4 mL Ringer solution or EP solution per 12 hours after PSH 12 till PSH 48. Rats in SS group were treated the same as that of S group except for sham scald with 37 degrees C water. Injured rats were sacrificed at post scald day (PSD) 1, 3, 5, 7 (rats in NC group were also sacrificed), and CD4(+)CD25(+)Treg were isolated from spleen with magnetic-activated cell sorting method. The content of HMGB1 in spleen and IL-2 level in supernatant were determined with ELISA. The expression of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) on Treg was determined with flow cytometry, and the proliferation activity of T lymphocytes was also detected (recorded as absorbance value). Data were processed with analysis of variance among groups and independent samples t test.

RESULTS(1) Compared with that of rats in SS group and EPT group, the expression of splenic HMGB1 in S group increased significantly on PSD 1 through PSD 7 [peaked on PSD 1: (46.7 +/- 8.3) ng/mg protein]. (2) Compared with that in SS group, the expression of CTLA-4 in S group was enhanced significantly on PSD 1 through PSD 5 (with t value respectively 10.459, 12.051, 4.029, P < 0.05 or P < 0.01); while that in EPT group decreased significantly on PSD 1 through PSD 7 as compared with that from S group (with t value respectively 2.796, 9.913, 9.581, 10.022, P < 0.05 or P < 0.01). (3) Compared with that of rats in SS group, the proliferation activity of T lymphocytes in S group was markedly suppressed on PSD 1 through PSD 7 (nadir on PSD1: 0.167 +/- 0.059), and release of IL-2 was decreased significantly [nadir on PSD 5: (44 +/- 24) pg/mL]. T lymphocytes proliferation activity was restored and excretion of IL-2 increased in EPT group as compared respectively with that of S group at each time point.

CONCLUSIONSThe release of HMGB1 may stimulate splenic Treg to mature, thereby induce suppression of proliferation activity of T lymphocytes and immune function. EP can ameliorate immune dysfunction in animals with delayed resuscitation through inhibiting the synthesis and release of HMGB1.

Animals ; Antigens, CD ; metabolism ; Burns ; immunology ; CTLA-4 Antigen ; Cell Proliferation ; HMGB1 Protein ; metabolism ; Interleukin-2 ; metabolism ; Male ; Pyruvates ; pharmacology ; Rats ; Rats, Wistar ; Spleen ; cytology ; immunology ; T-Lymphocytes, Regulatory ; cytology ; immunology

OBJECTIVETo investigate the effects of extracellular signal-regulated kinase (ERKs) inhibition by AG126 on tissue tumor necrosis factor-alpha (TNF-alpha) expression and multiple organ dysfunction in rats with postburn Staphylococcus aureus sepsis and its potential signal regulating mechanism.

METHODSTo reproduce postburn sepsis model, male Wistar rats were inflicated with 20% total body surface area third-degree scald followed by Staphylococcus aureus challenge. 34 rats were randomly divided into four groups as follows: normal control group (n = 6), scald control group (n = 6), postburn sepsis group (n = 12), and AG126 treatment group (n = 10). Tissue samples from the liver, kidneys and lungs were collected to determine phosphorylated ERKs by Western blot analysis, and TNF-alpha mRNA expression as well as its protein levels.

RESULTSIt was revealed that phosphorylated ERKs in the liver, lungs, and kidneys from postburn septic animals were up-regulated rapidly at 0.5 - 2.0 hours, being 1.94-fold (P < 0.05), 2.86-fold (P < 0.01), and 1.41-fold at 2.0 hours compared to normal controls, respectively. Treatment with AG126 could significantly reduce phosphorylated ERKs in lung tissue by 70.6% (P < 0.01) at 2.0 hours postburn sepsis, and almost completely inhibited ERKs activation in the liver and kidneys at various time points. Meanwhile, both TNF-alpha mRNA and protein expressions in the liver, lungs, and kidneys were significantly decreased in AG126-treated group following septic challenge (P < 0.05 or 0.01). In addition, 2.0 hours after Staphylococcus aureus infection, treatment with AG126 markedly improved hepatic and renal function parameters, including serum ALT, AST, Cr, as well as BUN levels (P < 0.05 or 0.01), together with significant decrease in pulmonary myeloperoxidase activity, compared to those without AG126 treatment.

CONCLUSIONThese data suggested that ERKs signal transduction might be involved in the pathogenesis of systemic inflammatory response and multiple organ dysfunction in postburn gram-positive bacterial sepsis. Early treatment with AG126 could significantly down-regulate TNF-alpha mRNA expression as well as protein levels in vital organs and attenuate multiple organ dysfunction induced by scald injury combined with Staphylococcus aureus challenge.

Animals ; Blotting, Western ; Burns ; complications ; Enzyme Inhibitors ; pharmacology ; Gene Expression ; drug effects ; Kidney ; metabolism ; physiopathology ; Liver ; metabolism ; physiopathology ; Lung ; metabolism ; physiopathology ; Male ; Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism ; Multiple Organ Failure ; metabolism ; physiopathology ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Staphylococcal Infections ; etiology ; physiopathology ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; Tyrphostins ; pharmacology

OBJECTIVETo investigate the therapeutic value of inhibiting the expression of insulin-like growth factor-I receptor (IGF-IR) using picropodophyllin (PPP) by studying the effects on proliferative and metastatic potentials of human hepatocellular carcinoma (HCC) using an in vitro cultured cell system.

METHODSIGF-IR expression in human HCC cell lines (Bel-7404, Bel-7402, HepG2, and Huh-7) and human hepatocytes (L02) was assessed at baseline (pre-treatment) and after PPP treatment by western blotting. Changes in cell cycle were analyzed by flow cytometry and in cell viability by sulforhodamine B staining. Early apoptosis was detected by annexin-V/FITC and propidium iodide double-staining assay. Caspase-3/7 activity was suppressed by z-VAD-FMK and analyzed by homogeneous luminescence assay. Effects on cell motility were tested by wound-scratch test. Between-group differences were assessed by t-test or one-way analysis of variance.

RESULTSIGF-IR was markedly up-regulated in all HCC cell lines (vs. non-hepatoma hepatocytes). HCC cells with PPP-inhibited IGF-IR showed time-dependent decreases in cell motility and viability. After treatment with PPP for 24 hours, the proportion of HCC cells in G1 phase was 2.1% +/- 0.4%, in S phase was 11.0% +/- 0.7%, and in G2/M phase was 87.1% +/- 0.6%, and no healing was observed in the wound-scratch assay. The PPP treatment induced cell apoptosis, as evidenced by enhanced caspase-3/7 activity; the proportion of annexin-V+/PI- cells was significantly higher in the HepG2 cells than in the non-hepatoma hepatocytes (16.4% +/- 0.4% vs. 5.8% +/- 0.2%, t = 14.05, P less than 0.01). After z-VAD-FMK treatment, the apoptosis rate was significantly higher in the HepG2 cells than in the non-hepatoma hepatocytes (11.3% +/- 0.7% vs. 5.8% +/- 0.2%, t = 11.83, P less than 0.01).

CONCLUSIONIGF-IR is associated with proliferation, cell motility, and apoptosis of HCC cells, and may be a promising molecular target for HCC.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Podophyllotoxin ; analogs & derivatives ; pharmacology ; Receptor, IGF Type 1 ; metabolism