Objective : To explore the effects of piperlongumine ( PL) on the proliferation and apoptosis of triplenegative breast cancer MDA⁃MB⁃231 cells in vitro and in vivo as well as its possible molecular mechanism. Methods: MDA⁃MB⁃231 cells were treated with different concentrations of PL. MTT assay was performed to detect the level of cell proliferation. The apoptotic level was detected by flow cytometry. Western blot was performed to detect protein expression levels of proliferating cell nuclear antigen(PCNA) , B ⁃cell lymphoma⁃2 (Bcl⁃2) , cyclin⁃dependent kinase inhibitor 1A(CDKN1A/p21) , phosphorylated signal transducer and activator of transcription 3(p⁃STAT3), signal transducer and activator of transcription 3 (STAT3) , hypoxia inducible factor⁃1 α ( HIF⁃1α ) and Survivin. MDA⁃MB⁃231 cells were used to model tumor bearing nude mice and then PL was injected by intraperitoneal ( i.p. ) . The tumor protein expression levels of PCNA , p ⁃STAT3 , STAT3 , HIF⁃1 α and Survivin were detected by Western blot. Results : PL inhibited cell proliferation and induced apoptosis in a dose⁃dependent manner. PL down⁃regulated the protein expression levels of PCNA , Bcl⁃2 , p ⁃STAT3 , HIF⁃1 α and Survivin , and up⁃regulated the protein expression level of p21 . Furthermore , PL inhibited tumor growth and down⁃regulated the protein expression levels of PCNA , p ⁃STAT3 , HIF⁃1 α and Survivin in nude mice. Conclusion PL inhibits the proliferation of MDA⁃MB⁃231 cells in vitro and in vivo , and the underlying mechanism can be related to the negative regulation of STAT3/HIF⁃1 α pathway.

Objective: To investigate the expression of mannose binding lectin 2 (MBL2) in hepatocellular carcinoma(HCC) cell lines and its effects on the proliferation and prognosis. Methods: MBL2 expression levels in patients of different sexes and survival analysis of patients were subsequently performed using bioinformatics method. The mRNA and protein expression levels were detected with the qRT⁃PCR and Western blot. Cell proliferation was examined by MTT assay and cell clone formation assay;Flow cytometry was used to detect the cell apoptosis and cell cycle. Gene set enrichment analysis(GSEA) was used to analyze the signal pathways of MBL2 enrichment. Results: Bioinformatics analysis and qRT⁃PCR results showed that MBL2 was highly expressed in male HCC patients compared with female HCC patients , and was related to the prognosis of HCC patients. Compared with the control group ,over⁃expression of MBL2 inhibited the viability and clone formation rate of Huh7 cells and MHCC⁃97H cells;In addition , cell cycles were arrested and cell apoptosis increased. GSEA and Western blot results showed that MBL2 was enriched in the cell cycle signal pathway. Overexpression of MBL2 inhibited the expression of CDK4 protein and promoted the expression of P16 and BAX protein. Conclusion The expression of MBL2 is down regulated in HCC , which is related to the prognosis of male patients with HCC and participates in the process of cell proliferation. MBL2 may be a potential therapeutic target for male patients with HCC.

Objective: To study the characteristics of clinical diagnosis and treatment for 3 children with phytosterolemia. Methods: The different clinical manifestations of 3 children with phytosterolemia were retrospectively reviewed. The case 1 and case 2, who were 7 years and 2 months old twin sisters, hospitalized for frequent epistaxis and abdominal pain. The case 3, who was 5 years and 7 months old male, came to the hospital for cutaneous xanthoma. The phytosterol levels in serum of the children were analyzed by gas chromatography-mass spectrometry, and the second generation sequencing method was used to analyze the disease-causing gene. Sanger sequencing method was used to verify the ABCG5 gene mutation and parental source. Results: (1) The case 1 and case 2 showed moderate anemia, raised reticulocytes, total bilirubin and indirect bilirubin as well as splenomegaly. The blood smear showed that there were more irregular red blood cells, such as oral red blood cells, increased large/giant platelets, and ristomycin-induced platelet aggregation test was decreased. The urine routine examination indicated that there was bleeding in the urinary system. The results of blood lipid test were almost normal. The case 3 showed mild anemia with normal shape of erythrocyte and normal size of spleen. The large/giant platelets increased. The results of platelet aggregation test, bilirubin and urine routine examination were in normal range, but the levels of total cholesterol and low-density lipoprotein cholesterol increased significantly. (2) The levels of serum phytosterol were significantly increased in all the 3 children. (3) Two heterozygous mutations were detectable in ABCG5 gene of case 1 and 2 which were complex heterozygous mutation, i.e., c.9041G＞A and c.751C＞T. The variations were from their father and mother respectively. In case 3, only one homozygous mutation was detectable in ABCG5 gene which originated from their parents. Conclusion When the child showed increased large/giant platelets, hemolytic anemia, erythrocytosis or xanthoma of skin and rised total cholesterol and low-density lipoprotein cholesterol at first visit, the possibility of phytosterolemia should be considered. The blood phytosterol content and gene detection should be carried out as early as possible in order to treat early and improve prognosis.

Objective: To study the relation between the upper first molar root tips and the maxillary sinus floor in patients with different vertical facial patterns using cone-beam computed tomography (CBCT) and to explore the reference data for safe clinical orthodontic treatments. Methods: Three-dimensional images were reconstructed from CBCT data. The CBCT data from 120 adolescents and adults were divided into three groups (low-angle group, average-angle group, high-angle group) based on vertical facial type. For each subject, the distance from the maxillary sinus floor to the upper first molar root tips was measured, and the types of contacts were classified. ANOVA and LSD t tests were used for statistical comparisons and performed using SPSS 19.0. Results: Of the 120 samples, only 27% of the upper first molar root tips lost their contacts with the maxillary sinus floor, and the other 73% of the root tips contacted the sinus to different extents. Significant differences in the distances from the maxillary sinus floor to the upper first molar root tips were found for different vertical facial types (P < 0.05). The high-angle group had the lowest sinus floor, relative to the root tips, of the three adult groups (P < 0.05). In the adult group with a low angle, the measured value for the palatal root tips was the lowest and was significantly different from those in the other groups (P < 0.05). Conclusion Seventy-three percent of the upper first molar root tips contacted the maxillary sinus floor. The maxillary sinus floor tended to be lower relative to the first molar root tips in patients with a high-angle facial pattern than in others. The roots protruded into the sinus to a greater extent.

Objective: To explore the clinical manifestations, histopathological features, diagnosis, treatment and prognosis of Rosai-Dorfman&prime;s disease (RDD) in the maxillofacial region and to review the relevant literature in order to improve the understanding, diagnosis and treatment of oral and maxillofacial RDD. Methods : The clinical manifestations, histopathological features, diagnosis, treatment, and prognosis of a patient with RDD in the maxillofacial region admitted to Shenzhen People&prime;s Hospital were analyzed, and the literature was reviewed for analysis. Results: The clinical manifestations were palpable masses of 3.5 cm × 2.0 cm × 1.0 cm in the right cheek and 3.0 cm × 2.0 cm × 1.0 cm in the right submaxillary area, with clear boundaries, good mobility, medium and hard textures, respectively, no tenderness, smooth surfaces, and no obvious nodules. On contrast-enhanced and plain CT scans of the maxilla and neck, a diffuse soft tissue shadow was seen in the right maxillofacial region with an unclear boundary and uniform density, and the contrast-enhanced scan also showed moderate and uniform enhancement. The primary diagnosis was right maxillofacial lesions. The tumor was resected surgically. The pathological report was right buccal and right submaxillary extranodal RDD. Under light microscopy, nodular lesions in the fibrous fat tissue were found, which were composed of light and deep staining areas. The light staining areas consisted of patchy, polygonal cells with large volumes and rich cytoplasm, in which lymphocytes and neutrophils could be seen stretching into the movement; the deep staining areas were composed of lymphocytes and plasma cells. IHC: S-100 (+), CD68 (+), CD163 (+), CD1a (-), CD21FDC (+), langerin (-), IgG (+), IgG4 (+). No recurrence was found 11 months after the operation. RDD is a rare, benign and self-limited tissue and cell disease and consists of multiple lesions in the maxillofacial region. Its imaging features are similar to those of lymphoma. Its pathological features are large volumes, rich cytoplasm and phagocytosis of lymphocytes and plasma cells. Generally, RDD only needs to be observed, and individuals with symptoms or the involvement of important organs need to be treated; the first choice for the extranodal type is drug treatment, with radiotherapy administered if the central nervous system is involved. Surgery is recommended if involvement of important organs and compression of the trachea are observed; chemotherapy should be used for diffuse RDD. Most patients with RDD experienced relapse or remission of the disease; a few patients died because of the involvement of important organs or complications. Conclusion The clinical manifestations of maxillofacial RDD vary and lack specific imaging features, and pathological immunohistochemistry is the gold standard for diagnosis. The etiology is not completely clear, the treatment methods are varied, and the prognosis is related to the involved range of the disease.

Objective: To observe the treatment effect of oral propranolol combined with topical timolol maleate for infantile maxillofacial mixed hemangioma and provide evidence for clinical treatment. Methods: Ninety-seven cases of infantile maxillofacial mixed hemangioma were enrolled. The cases were randomly divided into A and B groups: 50 cases in group A were treated with oral propranolol combined with topical timolol maleate, and 47 cases in group B were treated with oral propranolol only. The changes in the color, volume, and texture of the tumors were recorded before and after treatment, and color ultrasonography of the lesion area was performed. The follow-up time was 1-12 months. The differences in the curative effect, effective time and adverse reaction between the two groups were compared. Results : The effective rate of group A was 92.0% (46/50) and that of group B was 74.5% (35/47), with a statistical significance (P < 0.05). The mean time of treatment in group A was 4.2 months and that in group B was 5.5 months. Compared with group B, the treatment time of group B was shorter (t=3.211, P < 0.05), and no serious adverse reactions occurred in both groups. Conclusion Oral propranolol combined with topical timolol maleate is effective in the treatment of mixed hemangioma of the maxillofacial region in infants.

Objective: Abstract: Objective: To investigate the expression and significance of miR-138 and programmed cell death protein 1 (PD-1) in the patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). Methods: A total of 30 patients with HBV-related HCC, 20 with HBV-related cirrhosis (LC) and 30 with chronic hepatitis B (CHB) were recruited from Jiaozuo People&prime;s Hospital. The blood samples from all patients and the peritoneal effusion samples from HCC and LC patients were collected. The levels of miR-138 and soluble PD-1 (sPD-1) in blood and peritoneal effusion samples were detected by real-time PCR and ELISA, respectively. The expressions of PD-1 in T lymphocytes were measured with flow cytometry and western blot. The targeting effect of miR-138 on the 3&prime;-non-coding region (3&prime;-UTR) of PD-1 gene was verified by the dual-luciferase reporter gene system. Results: The relative expression levels of miR-138 in the peritoneal effusion and plasma of HBV-related HCC patients were significantly lower than those in LC and CHB patients (P＜0.05). The serum sPD-1 levels and the expression levels of PD-1 in CD3 + T lymphocytes of HBV-related HCC patients were significantly higher than those in LC and CHB patients (P＜0.05). The relative expression levels of miR-138 were negatively correlated with serum sPD-1 levels and the expression levels of PD-1 in CD3 + T lymphocytes (P＜0.05). The dual-luciferase reporter gene system and western blot results demonstrated that there was a targeting relationship between miR-138 and the 3&prime;-UTR of PD-1 gene. After miR-138 was transfected, the expression level of PD-1 was significantly down-regulated. Conclusion miR-138 participates in the development and progression of HBV-related HCC probably by targeting PD-1.

Objective : To investigate the effects of D ⁃allose on the restoration of neurological function , Galectin⁃3 (Gal⁃3) , adenosine monophosphate⁃activated protein kinase/mammalian target of rapamycin (AMPK/mTOR) and the expression of some inflammatory factors in ischemia⁃reperfusion injury ( CIRI) mice . Methods : A total of 50 male mice were randomly divided into control group (Con group) , sham group (Sham group) , cerebral ischemia⁃reperfusion injury group (MCAO group) , cerebral ischemia⁃reperfusion injury + D ⁃alolose group (MCAO + D ⁃allose group) and cerebral ischemia⁃reperfusion injury + modified citrus pectin group (MCAO + MCP group) . The middle cerebral artery occlusion/reperfusion (MCAO/R) model (reperfusion after 2 hours of MCA ischemia) was established by thread embolism . After successful modeling , the neurological function of mice was evaluated Longa score and rotated rod walking . TTC staining was used to observe the volume of cerebral infarction foci . The expression levels of Gal⁃3 and autophagy⁃related molecules were detected by Western blot and RT⁃PCR . Immunofluorescence was applied to detect the distribution of Gal⁃3 in brain tissue , and TNF⁃α , IL⁃8 secretion was detected with ELISA KIT . Results : Compared with Con group and Sham group , the MCAO model represented significant increase in the Longa neurofunction score (P < 0. 01) , cerebral infarction volume ( P < 0. 01) , Gal⁃3 expression and manifasted enhanced autophagy (P < 0. 01) . After treatment with D ⁃allose , it could significantly improve neurological dysfunction , reduce cerebral infarction volume (P < 0. 01) , reduce the expression of Gal⁃3 ( P < 0. 01) , inhibit AMPK phosphorylation , promote mTOR phosphorylation , and inhibit autophagy (P < 0. 01) . The use of the Gal⁃3 inhibitor MCP alone could also achieve the effect of inhibiting autophagy . Conclusion D ⁃allose can effectively promote the recovery of neurological function and reduce the volume of infarct foci in CIRI mice . The mechanism may involve inhibiting excessive cell autophagy by downregulating the expression of Gal⁃3 , and reducing the release of inflammatory factors such as TNF⁃α and IL⁃8 , thereby exerting neuroprotective effects .

Objective: To explore the relationship between MSX1 gene detection and tooth loss in a Van der Woude syndrome (VWS) family Methods : DNA was extracted from the venous blood of 2 patients with dental hy⁃podontia in the 9th family of Van der Woude syndrome (VWS) families and 62 controls with complete dentition. Primers were designed for the MSXl gene. The coding regions of exons 1 and 2 of the MSX1 gene were amplified by PCR. The purified products of exons 1 and 2 of the MSX1 gene were sequenced and analyzed by sequence alignment Results: The ivs2+68 C>T polymorphism in the MSX1 gene was found in the VWS9 members with tooth loss, and the VWS pa⁃tients with IRF6 gene mutations had increased tooth loss Conclusion Congenital tooth loss in the patients with con⁃genital missing teeth in VWS family 9 may be related to the ivs2 + 68 C> T polymorphism of the MSX1 gene.

Objective : To compare and analyze the effects of thermal etching on the shear strength of zirconia substrates and decorative ceramics. Methods: A total of 20 specimens made with zirconia ceramics were randomly divided into an observation group and a control group with 10 cases in each group. The control group was treated with sandblasting, while the observation group was treated with sandblasting and thermal etching. The surface characteristics were examined by scanning electron microscope (SEM) and phase analysis of X-ray diffraction (XRD), and the shear strength was tested using a universal testing machine. The characteristics of surface destruction were examined by SEM. Results : SEM showed that the peak structure was observed in both groups. The observation group exhibited deep fissures, and the control group exhibited small fissures. The diffraction peaks of the two groups are similar. The T (101) peak is the main peak, and both groups exhibit an M (111) peak. However, the peak intensity is relatively small. The relative levels of monoclinic zirconia were 15.16% in the observation group and 16.22% in the control group. The shear bond strength of the observation group was 24.74 ± 3.02 MPa, which was significantly higher than that of the control group at 21.09 ± 2.58 MPa. The difference was statistically significant (t=2.599, P=0.021). In the control group, the porcelain residue on the zirconia surface was minimal at low magnification, and the zirconia substrate was obviously exposed. The zirconia surface was similar to cristae obliqua at high magnification, and the porcelain exhibited a scattered distribution. In the observation group, a large amount of residual veneer porcelain remained on the zirconia surface at low magnification, but considerable porcelain was observed at high magnification. Conclusion Thermal etching and sandblasting treatment can improve the shear strength of zirconia substrate.