OBJECTIVE To explore the material basis of the anti-inflammatory effect of the petroleum ether extract from Melastoma dodecandrum by establishing its fingerprint and combining it with cellular pharmacodynamics experiments. METHODS HPLC method was adopted； the fingerprints of 20 batches of petroleum ether extract from M. dodecandrum were drawn using The Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）； similarity evaluation and common peak identification were carried out. The lipopolysaccharide-induced inflammation model of mice mononuclear macrophages （RAW264.7） was established； the inhibitory rates of nitric oxide （NO） and tumor necrosis factor α （TNF-α） were used as indexes to investigate the anti-inflammatory activity of the petroleum ether extract from M. dodecandrum； grey correlation degree method and partial least square regression analysis were adopted to study the spectrum-effect relationship. Molecular docking was used to validate the binding activity of the anti-inflammatory active ingredients with TNF-α and iNOS protein receptor. RESULTS There were 19 common peaks in the fingerprint of the petroleum ether extract from M. dodecandrum， the similarity of 20 batches of samples ranged from 0.603-0.990， and five components were identified， such as vitexin （peak 5）， isovitexin （peak 6）， ellagic acid （peak 7）， quercetin （peak 9） and luteolin （peak 10）. The grey correlation degree between 19 common peaks of the petroleum ether extract from M. dodecandrum and the inhibition rates of NO and TNF-α were all greater than 0.7； peaks 19， 13， 9 （quercetin）， 12， 5 （vitexin）， 6 （isovitexin）， 8， 7 （ellagic acid）， 18， 1 were positively correlated with NO inhibition rate， and peaks 8， 10 （luteolin）， 13， 15， 3， 19， 17， 7 （ellagic acid）， 18， 1 were positively correlated with inhibition rate of TNF-α. The binding energies of vitexin， isovitexin and quercetin with iNOS protein receptor were less than －5.0 kcal/mol. CONCLUSIONS Vitexin， isovitexin and quercetin may be the basis of the anti-inflammatory effect of the petroleum ether extract from M.dodecandrum.

Irritable bowel syndrome （IBS） is a functional gastrointestinal disease， but it often causes extreme gastrointestinal discomfort and prolonged illness， which seriously affects the quality of life of patients. The global incidence rate is increasing year by year. Clinically， western medicine mainly uses oral antispasmodics， secretagogues， and antidepressants， but there are many disadvantages such as adverse reactions and poor long-term efficacy. Therefore， finding an efficient and safe treatment method is an urgent problem to be solved. A large number of studies have shown that traditional Chinese medicine has definite curative and long-lasting effects on the treatment of IBS， which has become a hot research direction in recent years. By searching Chinese and foreign literature， it is found that electroacupuncture， moxibustion， Chinese medicine monomers， and compound decoctions are the main methods in the mechanism research of traditional Chinese medicine in the treatment of IBS-related pathways， and their signaling pathways involve nuclear transcription factor kappa B （NF-κB）， transient receptor potential vanillin subfamily 1 （TRPV1）， 5-hydroxytryptamine （5-HT）， mitogen-activated protein kinase （MAPK）， and so on. Traditional Chinese medicine can repair intestinal inflammation， reduce visceral sensitivity， enhance intestinal mucosal barrier， and regulate intestinal motility by regulating this series of signaling pathways， thereby playing an important role in the treatment of IBS with multi-level， multi-link and multi-target characteristics. Based on the cell signaling pathways， this paper reviewed the research progress on the mechanism of traditional Chinese medicine in the treatment of IBS， hoping to provide theoretical support and diagnosis and treatment ideas for the clinical treatment of IBS with traditional Chinese medicine.

Functional dyspepsia (FD) is a common and frequently occurring disease in clinic. With the influence of environmental factors, social factors and dietary factors, the incidence rate of FD in the general population is yearly increasing. Traditional Chinese medicine has a long history and far-reaching influence in the treatment of FD. It can prevent and treat FD in the form of multiple-components, targets and channels, with obvious effect and prominent advantages. This article starts with the common syndrome types of FD, and discusses the research progress of single Chinese medicine, effective ingredients and the mechanism of traditional Chinese medicines in treating FD, in order to provide a theoretical basis for the treatment of FD with traditional Chinese medicines.

Objective Colon-targeting capsules based on gastric pellets and enteric pellets were prepared from Baizhu Huanglian prescription. The formulation composition and preparation process were optimized and the in-vitro release characteristics were investigated. Methods Optimum formulation composition and process parameters of Baizhu Huanglian pellets were screened out by single factor experiment and orthogonal design. The pellets core were prepared by extrusion-spheronization technique and coated in the fluid bed using bottom spray coating technique. To investigate the effect of coating level of the isolation layer, the proportion of polymer, the amount of plasticizer and weight gain of enteric coating on the release behavior of the enteric pellets. The pellets release behavior was fitted by model as well. Results The prescription of gastric pellets was drug loading 50%, PVPP 5%, MCC to lactose 1∶2 and wetting agent 40%. The process parameters were extrusion frequency 20 Hz, rounding speed 500 r/min and rounding time 5 min. The prescription of enteric pellets was drug loading 27%, PVPP 5%, MCC to lactose 5∶2, wetting agent 30% and adhesive 20%. The process parameters were extrusion frequency 20 Hz, rounding speed 700 r/min and rounding time 7 min. For enteric coating layer, the coating mixture of EUDRAGIT®L30D-55 to EUDRAGIT® FS30D was 1∶2. The amount of plasticizer was 10%. The increased weight of coating layer was 15%. The release time of enteric pellets in-vitro was up to 24 hours. The release behavior of the pellets conforms to the Higuchi model. Conclusion The colon targeting capsule of Baizhu Huanglian pellets were successfully prepared and showed the characteristics of sustained release and colon targeting.

Objective: To investigate the expression of HOPX gene in cervical cancer tissues and blood serum as well as its effect on cervical cancer HeLa cells, and to analyze its correlation to tumor maker CEAand CA125. Methods: 50 pairs of cervical cancer tissues and para-cancerous tissues as well as the peripheral blood samples from patients with cervical cancer, who were treated at Tianjin Binhai People’s Hospital and Tianjin Wuqing People’s Hospital from June 2015 to December 2017, were collected for this study; in addition, 50 samples of blood serum from healthy people were used as control. Real-time quantitative PCR (qRT-PCR) and immumohistochemical staining (IHC) were used to detect mRNA and protein expressions of HOPX in tissue and serum samples, NCBI-GEO data base and TCGA data base were used to collect the information on HOPX gene and patients’prognosis, and the correlation between HOPX expression and patients’prognosis was analyzed. Vectors over-expressing HOPX or control vectors were transfected into HeLa cells; MTT assay and colony formation assay were used to examine the proliferation ability of HeLa cells, Tranwell assay was used to detect the migration and invasion of HeLa cells, and Western blotting was used to detect the expression of EMT-related proteins. Results: Both sample examination and data base information showed that the expression level of HOPX was down-regulated in tissue and serum samples of cervical cancer patients and was positively related with the survival of patients (r=0.736， P<0.05); while it’s expression was negatively related to the level of CEAand CA125 in cervical cancer tissues and serum (r=-0.678, P<0.05). HOPX over-expression inhibited cell proliferation, migration and invasion, promoted the expression of E-cadherin but inhibited the expression of Vimentin and ICAM1 (all P<0.05 or P<0.001). Conclusion: HOPX is low expressed in cervical cancer tissues and blood samples, and negatively correlated with CEA and CA125, but positively correlated with the survival of patients. Thus, combination of HOPX and CEA/CA125 may improve the early diagnosis rate of cervical cancer and provide a new strategy for precision treatment of cervical cancer in future.

BACKGROUND:Nowadays, most of the studies regarding tissue engineering bone have mostly focused on critical-size bone defects of the backbone; however, there are less studies and reports on its spinal fusion. OBJECTIVE:To explore the feasibility of xenogeneic deproteinized cancelous bone as bone tissue engineering scaffold in the treatment of spinal intertransverse fusion. METHODS:The cancelous part in the distal femur of adult pigs was obtained to prepare xenogeneic deproteinized cancelous bone. After combined with the recombinant human bone morphogenetic protein, the xenogeneic deproteinized cancelous bone was combined with bone marrow mesenchymal stem cels to prepare tissue engineering bone. Twenty-four goats were obtained to prepare intertransverse bone bed, and randomly divided into two groups: observation and control groups. In the observation group, the tissue engineered bone was implanted into the left side, and the xenogeneic deproteinized cancelous bone of recombinant human bone morphogenetic protein was implanted into the right side. In the control group, the autologous iliac bone was implanted into the left side, and xenogenic deproteinization cancelous bone was implanted into the right side. At the 4th, 8th and 12th weeks after implantation, the fusion segment was obtained for gross observation, X-ray observation, histological observation and biomechanical testing. RESULTS AND CONCLUSION:X-ray films showed that the implant materials from these two groups were fixed wel and reliably. At different time points after implantation, the implant materials from each group were al in good position. There were no purulent and necrotic tissues around the material. Soft tissue ingrow and wraping were present. There were no effusions and necrosis surrounding the implant materials. The imaging and histological performance in the tissue engineering bone group outperformed that in the recombinant human bone morphogenetic protein xenogenic deproteinized cancelous bone group and xenogenic deproteinized cancelous bone group, which was the most close to the autogenous bone. At the 12th week after implantation, the maximum bending load in the tissue engineering bone group was the most close to the autogenous iliac bone group. There was no significant difference between these two groups. These results demonstrate that as bone tissue engineering scaffold, xenogenic deproteinized cancelous bone has a certain application feasibility in the treatment of spinal intertransverse fusion.

Objective To investigate the expression of CXCR7 in several cutaneous malignant tumors including cutaneous squamous cell carcinoma (SCC),basal cell carcinoma (BCC) and invasive cutaneous malignant melanoma and their cell lines,as well as its significance.Methods Tissue specimens were obtained from the lesions of 30 patients with cutaneous squamous cell carcinoma,25 patients with basal cell carcinoma and 30 patients with cutaneous malignant melanoma.Immunohistochemistry was performed to detect the expression of CXCR7 protein in these tissue specimens and several cell lines (A375 human melanoma cells,M14 human melanoma cells,A431 human epidermoid carcinoma cells,HaCaT human keratinocytes).The mRNA expression of CXCR7 in these cell lines was measured by reverse transcription PCR.Results CXCR7 protein was apparently expressed in invasive cutaneous malignant melanoma.The high expression rate of CXCR7 protein was significantly elevated in cutaneous malignant melanoma tissue specimens compared with SCC and BCC tissue specimens [80％ (24/30) vs.26.67％ (8/30) and 8％ (2/25),x2 =17.16,28.36,both P ＜ 0.05].CXCR7 mRNA was expressed in A375,M14 and A431 cells,but not in HaCaT cells,with the strongest expression observed in A375 cells.Immunohistochemistry revealed the expression of CXCR7 protein only in A375 cells.Conclusions CXCR7 is highly expressed in cutaneous malignant melanoma and A375 cells,which may be involved in the malignant invasion and metastasis of melanoma.

OBJECTIVE: To investigate the expression and clinical significance of MMP9 and MVD in the carcinogenesis of squamous cell carcinoma of external auditory canal and middle ear. METHOD: Immunohistochemical SP method was used to detect the expression of MMP9 and MVD proteins in 26 squamous cell carcinoma tissues of external auditory canal and middle ear and 20 normal external ear canal skin tissues. RESULT: The positive rate of MMP9 in squamous cell carcinoma tissues of external auditory canal and middle ear was 73.1% (19/26) lower than that in the normal external ear canal skin tissues 25.0% (5/20). The positive rates of CD34 were 33.58 +/- 3.04 and 22.50 +/- 5.22, respectively. The positive rates of MMP9 and CD34 were correlated with the histological grade and tumor grade, but had no relationship with age and sex. The positive rates between MMP9 and CD34 were related (r=0.42, P<0.05). CONCLUSION MMP9 may be involved in the carcinogenesis of squamous cell carcinoma of external auditory canal and middle ear, and may play an important role in the invasion and metastasis of squamous cell carcinoma of external auditory canal and middle ear. MMP9 and CD34 play a cooperative role in the process of squamous cell carcinoma of external auditory canal and middle ear.
Adult ; Aged ; Antigens, CD34 ; metabolism ; Carcinoma, Squamous Cell ; blood supply ; metabolism ; Ear Canal ; metabolism ; Ear Neoplasms ; blood supply ; metabolism ; Ear, Middle ; metabolism ; Female ; Humans ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Microvessels ; pathology ; Middle Aged ; Neovascularization, Pathologic

OBJECTIVETo compare effects of vinblastine (VLB) nanoparticles (NPS) and VLB physiologic saline solution on inhibiting glioma cell lines C6 growth and inducting its apoptosis.

METHODGlioma cell lines C6 were respectively treated with 500 micro x L(-1) VLB NPS and VLB physiologic saline solution for 7 days. Amount of cells were counted by blood cell counting chamber. Glioma C6 growth curve was draw according to cells amount. Clone formation rate of glioma C6 was detected after 500 microg x L(-1) VLB NPS and VLB physiologic saline solution incubation for 2 weeks. In addition, the whole morphology of glioma C6 were observed by inverted microscope and inverted fluorescence microscope after 500 microg x L(-1) VLB NPS and VLB physiologic saline solution incubation for 48 h.

RESULTEntrapment of VLB in NPS may significantly inhibit glioma cells C6 growth from 2 to 7 days compared with VLB physiologic saline solution in the same dose (P < 0.05). Clone formation rate of glioma C6 in VLB physiologic saline solution group is 1. 3 times better than VLB NPS. The difference between VLB NPS and VLB physiologic saline solution is significant (P < 0.05). Results of morphology change indicated glioma cells C6 with the VLB NPS treatment were intermediate or end stage, missed structure integrality. Amount of cells was distinctly decreased, and apoptosis cells number was apparently increased compared with VLB physiologic saline solution group.

CONCLUSIONVLB NPS have stronger cytotoxicity to glioma cells line C6 compared with VLB physiologic saline solution in the same dose. NPS may be effective as promising carrier for the transport of VLB into the glioma cells.

Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Glioma ; drug therapy ; physiopathology ; Nanoparticles ; chemistry ; Rats ; Vinblastine ; pharmacology

Objective To investigate the effect ofpeptidoglycan from Staphylococcus aureus on the release of several chemokines including intedeukin 8 (IL-8), regulated upon activation, normal T cell expressed and secreted (RANTES), macrophage-derived chemokine (MDC) by normal human epidermal keratinocytes (KCs) and the role of Toll-like receptor 2 (TLR2) in this process. Methods KCs were derived from the foreskin of a healthy boy and propagated. After 2 - 4 passages, KCs were collected and treated with various concentrations (3, 10, 30 and 100 mg/L) of peptidoglycan for 24 hours or with peptidoglycan of 100 mg/L for varying durations (3, 6, 12, 36 hours). A fi'action of KCs were pretreated with functional grade purified anti-TLR2 monoclonal antibody before the treatment with peptidoglycan of 100 mg/L. After additional 12-hour culture following the treatment, enzyme linked immunosorbent assay was used to detect the level of IL-8, RANTES and MDC in culture supernatants of KCs. Results KCs spontaneously released IL-8 and RANTES. Peptidoglycan increased the production of IL-8 but decreased that of RANTES by KCs. The levels of IL-8 were 209.96 ± 10.31 ng/L, 250.28 ± 9.52 ng/L, 285.11 ± 10.28 ng/L, 359.40 ± 6.93 ng/L in KCs treated with peptidoglycan of 3, 10, 30, 100 mg/L, respectively, compared to 135.41 ± 14.37 ng/L in untreated KCs (all P < 0.05). On the contrary, a significant decrement was seen in the secretion of RANTES by KCs treated with peptidoglycan of 10, 30, 100 mg/L compared with untreated KCs (110.72 ± 8.51 ng/L, 90.50 ±2.45 ng/L, 49.89 ± 13.74 ng/L vs 149.94 ± 18.71 ng/L, all P < 0.05). The monoclonal antibody to TLR-2 could markedly suppress the promotion of IL-8 production by peptidoglycan, but had no obvious influence on the inhibition of RANTES production by peptidoglycan. MDC could not be detected in the culture super-natants of KCs with or without peptidoglycan stimulation. Conclusion Peptidoglycan could inhibit RANTES secretion but induce IL-8 production by KCs likely via TLR2.