OBJECTIVE:To determine content of paeoniflorin in shaofuzhuyu superfine powder granules by HPLC. METHODS: The separation was performed on C18 chromatographic column, the mobile phase was methanol -0.05mol/L KH2PO4(40∶65)with a flow speed at 1ml/min, detection wavelength at 230nm and column temperature at 30℃. RESULTS:The linear range of paeoniflorin was 15.04～752.00?g/ml(r=0.9 998,n=6),the mean recovery rate was 103.3%(RSD=1.53%,n=3).CONCLUSION:This method was simple, accurate, good in recurrence and high in precision, and which can be used for the contents determination and the quality control of the shaofuzhuyu superfine powder granules.

Objective To observe the distribution of sentinel nodes(SN) in gastric cancer,and investigate their clinical significance.Methods The extent and features of intraoperatively stained sentinel nodes in 288 cases of gastric cancer were analyzed retrospectively.Methylene blue was injected intraoperatively into the(subseral) muscle and submucosa of the normal gastric wall adjacent to the neoplasms,then lymphadenography was performed,and the nodes were resected for histopathological examination.Results The pathological(diagnosis) of the 288 cases with gastric cancer was T_1 in 102 cases,T_2 106 cases and T_3 60 cases.Stained nodes were detectable in 270 cases(success rate: 93.8%).One hundred and two cases were found to have lymph node metastasis;66 cases had metastasis in both SNs and non-SNs;18 cases in SNs alone;and 18 cases in non-SNs alone.Conclusions Sentinel lymph nodes can accurately predict the lymph node metastatic status of gastric cancer patients during operation.Intraoperative sentinal lymph node mapping can avoid routine lymph node dissection in patients with gastric cancer with negative lymph node metastasis.

Cadherins ; genetics ; Hearing Loss, Sensorineural ; genetics ; pathology ; Humans

Objective To investigate the intra-laboratory turnaround time(ILTAT) of the emergency biochemistry tests and to analyze the factors influencing ILTAT in order to adopt the corresponding improvement measures for perfecting the service quality and ensuring the patient medical safety.Methods ILTAT of the emergency biochemical specimens in our hospital from June to November 2015 was performed the retrospective statistics for comparing the determination timely rate between ILTAT≤60 min and ILTAT2 ≤120 min.ILTAT at different time periods in laboratory was emphatically analyzed.Results The determination timely rate of ILTAT ≤120min(ILTAT 1) was 98.8%(8638/8743),and which of ILTAT ≤60min(ILTAT 2) was 83.7%(7317/8743).The determination timely rate of ILTAT1 had no statistical difference among different time periods (χ2=3.36,P>0.05).The determination timely rate of ILTAT2 had statistical difference among different time periods(χ2=134.5,P<0.01).The determination timely rate of T 2(10:01-12:00) was highest (88.1%),which of T1 (8:01-10:00) was lowest(76.8%),which of T3(12:01-14:00) and T7 (6:01-8:00) was lower (79.4% and 80.2% respectively).Conclusion At present,ILTAT in our laboratory meets the requirements of the current regulations.Analyzing the ILTAT influencing factors in the emergency biochemistry,optimizing the workflow,improving the equipments and staffing allocation and improving the degree of information processing can further shorten the emergency biochemical ILTAT,and better meet the clinical and patient′s needs.

Objective:To evaluate the effect of ribosomal protein L34 (RPL34) gene knockdown on the proliferation and apoptosis of human cutaneous squamous cell carcinoma (cSCC) cells.Methods:From January 2016 to January 2017, 14 paraffin-embedded skin samples of cSCC and 16 paraffin-embedded normal skin tissue samples were collected from Department of Dermatology and Venereology, the Affiliated Hospital of Inner Mongolia Medical University, and RPL34 expression in the skin tissues was analyzed by immunohistochemical study. A lentivirus vector containing short hairpin RNA targeting RPL34 gene was constructed and used to transfect a human cSCC cell line SCL-1 (shRNA group) , SCL-1 cells transfected with an empty lentivirus vector served as control group, and the knockdown efficiency was verified by real-time quantitative PCR (RT-PCR) and Western blot analysis. At 72 hours after the transfection, flow cytometry was performed to analyze the cell cycle and detect apoptosis of SCL-1 cells, and methyl thiazolyl tetrazolium (MTT) assay to evaluate the cellular proliferative activity of SCL-1 cells. Comparisons between 2 groups were performed by using t test or rank sum test. Results:Immunohistochemical study showed that the cytoplasmic expression score of RPL34 was significantly higher in the cSCC tissues (2.143±1.956) than in the normal control tissues (0.500±0.516, z=3.53, P< 0.05) . RT-PCR showed that the relative mRNA expression of RPL34 in the SCL-1 cells was significantly lower in the shRNA group (0.149±0.016) than in the control group (1±0.018, t=36.95, P< 0.05) ; Western blot analysis revealed that the relative protein expression of RPL34 in the SCL-1 cells was significantly lower in the shRNA group than in the control group. Compared with the control group, the shRNA group showed a significantly increased proportion of S-phase cells ( t=13.76, P< 0.05) , but a significantly decreased proportion of G1-phase cells ( t=36.62, P< 0.05) ; the apoptosis rate was significantly higher in the shRNA group (9.42%±0.16%) than in the control group (4.58%±0.41%, t=19.02, P< 0.05) . MTT assay showed that the cell viability was significantly decreased in the shRNA group (0.815±0.005) than in the control group (1.886±0.005, t=265.91, P< 0.05) after additional 120-hour culture. Conclusion:The RPL34 gene was overexpressed in the cSCC tissues, and knockdown of the RPL34 gene in SCL-1 cells could interfere with cell cycle, decrease their proliferative activity, and promote their apoptosis.

Methanotrophs are a group of bacteria capable of utilizing methane as the sole carbon and energy source for their anabolism and catabolism. Since methanotrophs contain the unique enzymes of methane monooxygenases (MMOs), which can catalyze the oxidation of methane and short-chain alkanes and alkenes, they have potential applications in carbon recycle of nature and industrial biotechnology. Therefore, methanotrophs have been paid much more attention by the researchers in recent 20 years. In this paper, the latest progresses in studies of methanotrophs and MMOs were reviewed, including taxonomy, function and distribution of methanotrophs, and structure, function and genetic engineering of MMOs. The future research directions of methanotrophs and MMOs as well as their applications were also discussed.
Methane ; metabolism ; Methylococcaceae ; enzymology ; genetics ; Oxidation-Reduction ; Oxygenases ; metabolism

OBJECTIVETo investigate the influence of early enteral nutrition with synbiotics on the plasma endotoxin level, the nutritional state, the inflammatory response and the incidence of infectious complications in severely burned patients.

METHODSRandomized double blind and control method was employed im the study. Forty severely burned patients were randomly divided into A and B groups with 20 in each group. The patients in group A received early enteral nutrition with synbiotics including four kinds of lactic acid bacteria and four kinds of fibers, while those in group B received early enteral nutrition with synbiotics including only four kinds of fibers. The patients with 80% to 280% coefficient unit burned surface(UBS) were further divided into A1 (n = 10) and B1 (n = 11) groups. The plasma endotoxin level in group A and B was determined dynamically on the 1st, 3rd, 7th, 10th, 14th, and 21st postburn days (PBD), and its abnormal rate in both groups was statistically analyzed in correlation with the normal endotoxin level. meanwhile, the mortality, the incidence of infectious complications and the blood bacterial culture results were compared between the two groups. The plasma levels of IL-1, IL-6 and prognostic inflammatory nutrition index (PINI) were also determined at the above time points.

RESULTSThe plasma endotoxin level in group A (37.9 +/- 5.4) ng/L was evidently lower than that in group B (59.1 +/- 7.9) ng/L (P < 0.05) on 10 PBD. The abnormal rate of plasma endotoxin in group A (36.7%) was evidently lower than that (49.2%) in group B (P < 0.05). Blood culture was positive in 3 patients in group A, and 5 in group B. There was no obvious difference in the incidence of infectious complication between the two groups. Two patients died in group A and 1 group B. There was no obvious difference in plasma IL-1 level between A1 and B1 groups at different time points. The plasma IL-6 level in A1 group in 10th and 14th PBD was evidently lower than that in B1 group (P < 0.05). The PINI in A1 group on the 10 PBD was remarkably lower than that in B1 group.

CONCLUSIONEarly enteral nutrition with synbiotics was helpful in decreasing inflammatory stress response and lowering the plasma endotoxin level. Enteral supplementation of synbiotics might be beneficial to the controlling of burn infection.

Adult ; Burns ; therapy ; Dietary Fiber ; administration & dosage ; therapeutic use ; Double-Blind Method ; Endotoxins ; blood ; Enteral Nutrition ; methods ; Female ; Humans ; Intestinal Mucosa ; drug effects ; Male ; Probiotics ; administration & dosage ; therapeutic use

Angiogenic gene therapy and cell-based therapy for peripheral arterial disease(PAD) have been studied intensively currently. This study aimed to investigate whether combining mesenchymal stem cells(MSCs) transplantation with ex vivo human hepatocyte growth factor(HGF) gene transfer was more therapeutically efficient than the MSCs therapy alone in a rat model of hindlimb ischemia. One week after establishing hindlimb ischemia models, Sprague-Dawley(SD) rats were randomized to receive HGF gene-modified MSCs transplantation(HGF-MSC group), untreated MSCs transplantation (MSC group), or PBS injection(PBS group), respectively. Three weeks after injection, angiogenesis was significantly induced by both MSCs and HGF-MSCs transplantation, and capillary density was the highest in the HGF-MSC group. The number of transplanted cell-derived endothelial cells was greater in HGF-MSC group than in MSC group after one week treatment. The expression of angiogenic cytokines such as HGF and VEGF in local ischemic muscles was more abundant in HGF-MSC group than in the other two groups. In vitro, the conditioned media obtained from HGF-MSCs cultures exerted proproliferative and promigratory effects on endothelial cells. It is concluded that HGF gene-modified MSCs transplantation therapy may induce more potent angiogenesis than the MSCs therapy alone. Engraftment of MSCs combined with angiogenic gene delivery may be a promising therapeutic strategy for the treatment of severe PAD.
Animals ; Bone Marrow ; metabolism ; pathology ; Bone Marrow Transplantation ; Cells, Cultured ; Hepatocyte Growth Factor ; genetics ; Hindlimb ; pathology ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; pathology ; Neovascularization, Physiologic ; genetics ; Rats

OBJECTIVETo explore the pathological changes of pulmonary fibrosis induced by SiO2 in rats and pigs.

METHODSThe silicosis models in rats and pigs were established by non-exposure method. The pathologic changes in lung tissues of rats and pigs were observed with HE staining under a light microscopy and under a transmission electron microscopy (TEM), the expression of cytokines was detected by immunohistochemistry.

RESULTS(1) The main pathologic changes of silicosis models in rats and pigs included: in 7 ∼ 15 days after treatment, silica dusts, dust cells, a lot of macrophages, lung epithelial cells, a few neutrophils, macrophage alveolar inflammation and nodules of stage I were found in alveolar space; in 30 ∼ 90 days after treatment, many nodules of stage I-III or IV with lymphocytes infiltration were observed in respiratory bronchioles, alveoli, interlobular septa, the subpleural and around blood vessels and bronchi. (2) The expression levels of CK protein, SP-A protein, CD68, b-FGF, TNF-α, IL-6, TGF-β1, NFKappa/P50, Kappa/P65 and VEGF reduced with exposure time, but still were higher than those of the control. (3) The shed alveolar type I cells, proliferation of alveolar type II cells or macrophages and activated cellular function induced by silica were observed under TEM.

CONCLUSIONThe development of pulmonary fibrosis in silicosis models corresponded with the process from macrophages alveolar inflammation to pulmonary fibrosis.

Animals ; Cytokines ; metabolism ; Disease Models, Animal ; Epithelial Cells ; metabolism ; Female ; Lung ; cytology ; pathology ; Macrophages, Alveolar ; metabolism ; Male ; Neutrophils ; metabolism ; Rats ; Rats, Sprague-Dawley ; Silicosis ; pathology ; Swine

Methanotrophs use methane as the sole carbon and energy source, which cause slow growth, low cell density and hinder its industrial applications. One promising solution is to heterologously express methane monooxygenase (MMO) in other host cells that can be easily cultivated at high cell density. We systematically exploited the possibility of functional expression of pMMO by choosing different promoters and different host cells. The results showed that the recombinants could oxidize methane to methanol. In particular, ethanol could also be detected in the oxidized products, but the enzyme activity was instable, implying that some changes of pMMO expressed in the host cells might have occurred. In addition, SDS-PAGE analysis showed that many recombinants could express the subunits of pMMO, but the enzyme activity could not be detected. In conclusion, correct fold of pMMO in the host cells is important for its functional expression.
Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation, Enzymologic ; Genetic Vectors ; genetics ; Methane ; metabolism ; Methanococcaceae ; enzymology ; Methanol ; metabolism ; Oxygenases ; biosynthesis ; genetics ; Promoter Regions, Genetic ; Protein Folding ; Recombinant Proteins ; biosynthesis ; genetics