The aim of this study is to investigate the effects of Gynostemma pentaphyllum dried with two different methods(air drying and heating) on inflammation in acute lung injury(ALI) mice in vivo and in vitro. Lipopolysaccharide(LPS) was sprayed into the airway of wild type C57BL/6J male mice to establish the model, and the drug was injected into the tail vein 24 h after modeling. Lung function, lung tissue wet/dry weight(W/D) ratio, the total protein concentration, interleukin 6(IL-6), IL-1β, and tumor necrosis factor-α(TNF-α) in the bronchoalveolar lavage fluid(BALF), and pathological changes of the lung tissue were used to evaluate the effects of different gypenosides on ALI mice. The results showed that total gypenosides(YGGPs) and the gypenosides substituted with one or two glycosyl(GPs_(1-2)) in the air-dried sample improved the lung function, significantly lowered the levels of IL-1β and TNF-α in BALF, and alleviated the lung inflammation of ALI mice. Moreover, GPs_(1-2) had a more significant effect on inhibiting NO release in RAW264.7 cells. This study showed that different drying methods affected the anti-inflammatory activity of G. pentaphyllum, and the rare saponins in the air-dried sample without heating had better anti-inflammatory activity.
Male ; Mice ; Animals ; Tumor Necrosis Factor-alpha/metabolism* ; Gynostemma ; Mice, Inbred C57BL ; Lung ; Anti-Inflammatory Agents/metabolism* ; Interleukin-6/metabolism* ; Interleukin-1beta/metabolism* ; Lipopolysaccharides/pharmacology*

OBJECTIVE: To investigate the effect of silencing DNA methyltransferase 1(DNMT1) to the methylation of the promoter of the tumor suppressor gene wnt-1 (WIF-1) in human chronic myeloid leukemia (CML) cells. METHODS: DNMT1 siRNAi plasmid was constructed and DNMT1 siRNAi was transfected into CML K562 cells. RT-PCR and Western blot were used to detect the expression of DNMT1 gene and related protein, and methylation PCR was used to detect WIF-1 gene promoter methylation level. The trypan blue exclusion and MTT assay were used to detect the cell proliferation, flow cytometry were used to detect the cell apoptosis rate, colony formation assay was used to detect cell colony formation ability. Expression of Wnt/β- catenin and its downstream signaling pathway proteins were detected by Western blot after DNMT1 gene was silenced. RESULTS: The expression level of DNMT1 mRNA and its related protein in the experimental group were significantly lower than those in the control group and negative control group (P<0.05). After 72 hours of successful transfection, the WIF-1 gene in the control group and negative control group were completely methylated, while in the experimental group, the methylation level significantly decreased. The results of MSP showed that the PCR product amplified by the unmethylated WIF-1 primer in the experimental group increased significantly,while by the methylated WIF-1 primer decreased significantly. After 48 h of transfection, the OD value, viable cell number and colony formation of the cells in experimental group were significantly lower than those in the negative control group and the control group (P<0.05). The apoptosis rate of the cells in experimental group was significantly higher than those in the negative control group and control group (P<0.05). The expression levels of β- actin, myc, cyclin D1 and TCF-1 in K562 cells in the experimental group were significantly lower than those in the negative control group and control group (P<0.05). CONCLUSION Silencing DNMT1 gene can inhibit the proliferation and promote the apoptosis of K562 cells. The mechanism may be related to reverse the hypermethylation level of the WIF-1 gene promoter, thereby inhibit the activity of the Wnt/β- catenin signaling pathway.
Adaptor Proteins, Signal Transducing/metabolism* ; DNA Methylation ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics* ; Repressor Proteins/metabolism*

OBJECTIVE: To investigate the effects of high dose vitamin C on proliferation and apoptosis of acute myeloid leukemia (AML) cell lines including HL-60, U937 and primary CD34 leukemia cells in AML. METHODS: CD34 cells were sorted by using immunomagnetic cell sorting system, then the primary CD34 leukemia cells, including HL-60 and U937 cell lines were cultured in vitro. Cells in each group were treated with different concentrations of vitamin C, the survival rate of cells was determined by MTT assay, the apoptosis rate of cells was evaluated by Annexin V/PI double staining, the expression of apoptotic proteins-including cleaved caspase 3, cleaved caspase-9 and cleaved PARP were detected by Western blot. RESULTS: The proliferation of HL-60 and U937 cells could be inhibited by high dose vitamin C, which showed a concentration-dependent manner (r=-0.9664; r=-0.9796). HL-60 and U937 cells were treated with different concentrations of vitamin C (8 and 20 mmol/L) for 24 hours, respectively, it was found that with the increasing of vitamin C concentration, cell apoptosis rate was significantly increased (r=0.9905; r=0.9971), and the expression of apoptosis related proteins including cleaved caspase 3, cleaved caspase-9 and cleaved PARP was aslo significantly increased with the increasing of concentration. In addition, it was found that with or without the mutation of TET2, high dose vitamin C could inhibit the proliferation (r=-0.9719; r=-0.9699) and promote the apoptosis (r=0.9998; r=0.9901) of primary CD34 leukemia cells in AML, which showed a dose-dependent manner, but it showed no effect on the proliferation (r=-0.2032) and apoptosis (r=0.1912) of normal CD34 cells. CONCLUSION High dose vitamin C can inhibit the proliferation and promote the apoptosis of acute myeloid leukemia cells, and selectively kill primary CD34 leukemia cells in AML.
Apoptosis ; Ascorbic Acid ; Cell Proliferation ; HL-60 Cells ; Humans ; Leukemia, Myeloid, Acute ; U937 Cells

Objective:To assess the clinical outcome of the novel computer assisted reduction malarplasty using angled double L-shaped osteotomies.Methods:Retrospective analysis of the 35 female patients who received reduction malarplasty surgery during June 2014 to April 2019 was conducted. Patients were divided into the conventional surgery group (9 cases) and the computer assisted surgery group (26 cases) based on their personal will. For the conventional surgery group, the zygomatic arch was repositioned inwardly after L-shaped osteotomy, and was rigidly fixed with miniplates and screws. The computer assisted reduction malarplasty was as follows: computer assisted angled double L-shaped osteotomies with surgical guide was performed intraorally, and the pre-bent titanium was used to setback the resected zygoma bone, which was then fixed with titanium miniplates and screws. Operation time, patients’ satisfaction (3-month follow-up) and postoperative complications (asymmetry and bone nonunion) were recorded and assessed. CT scans were performed to compare the preoperative design and 3-month postoperative follow-up for the computer assisted patient group. For statistical analysis, independent sample t test was used to analyze operation time of the 2 groups and chi-square test was used to analyze the data of patients’ satisfaction and asymmetry occurrence. P<0.05 was considered as statistically significant. Results:The mean operation time was (85.1 ± 17.8) min during computer assisted surgery versus (62.2±11.7) min during conventional surgery. The difference between the two groups was statistically significant ( t=3.53, P=0.020). Neither group showed noticeable resected bone shifting or soft tissue drooping. One patient in the conventional surgery group had bone nonunion on the right zygoma and partial absorption of the left zygomatic bone. The incidence of asymmetry in the computer-assisted group was 3.8% (1/26, surgical correction was not required), and 33.3% in the conventional surgery group (3/9, one patient required surgical correction). There was a statistically significant difference between the two groups ( χ2=6.179, P=0.046). Patients’ satisfaction in the computer-assisted group was 100% (26/26), and 78% (7/9) in the conventional surgery group ( χ2=7.929, P=0.019). Comparisons between the postoperative CT and preoperative simulation CT images showed that the position deviation of the resected bones was (0.21 ± 0.19) mm. Conclusions:In the present study, improved precision of zygomatic bone resection and bone setback was achieved in reduction malarplasty by using the angled double L-shaped osteotomies with computer assistance. Moreover, complication occurrences (asymmetry, bone nonunion etc.) were significantly decreased. Also, patients’ expectation was better achieved with this method.

Objective:To assess the clinical outcome of the novel computer assisted reduction malarplasty using angled double L-shaped osteotomies.Methods:Retrospective analysis of the 35 female patients who received reduction malarplasty surgery during June 2014 to April 2019 was conducted. Patients were divided into the conventional surgery group (9 cases) and the computer assisted surgery group (26 cases) based on their personal will. For the conventional surgery group, the zygomatic arch was repositioned inwardly after L-shaped osteotomy, and was rigidly fixed with miniplates and screws. The computer assisted reduction malarplasty was as follows: computer assisted angled double L-shaped osteotomies with surgical guide was performed intraorally, and the pre-bent titanium was used to setback the resected zygoma bone, which was then fixed with titanium miniplates and screws. Operation time, patients’ satisfaction (3-month follow-up) and postoperative complications (asymmetry and bone nonunion) were recorded and assessed. CT scans were performed to compare the preoperative design and 3-month postoperative follow-up for the computer assisted patient group. For statistical analysis, independent sample t test was used to analyze operation time of the 2 groups and chi-square test was used to analyze the data of patients’ satisfaction and asymmetry occurrence. P<0.05 was considered as statistically significant. Results:The mean operation time was (85.1 ± 17.8) min during computer assisted surgery versus (62.2±11.7) min during conventional surgery. The difference between the two groups was statistically significant ( t=3.53, P=0.020). Neither group showed noticeable resected bone shifting or soft tissue drooping. One patient in the conventional surgery group had bone nonunion on the right zygoma and partial absorption of the left zygomatic bone. The incidence of asymmetry in the computer-assisted group was 3.8% (1/26, surgical correction was not required), and 33.3% in the conventional surgery group (3/9, one patient required surgical correction). There was a statistically significant difference between the two groups ( χ2=6.179, P=0.046). Patients’ satisfaction in the computer-assisted group was 100% (26/26), and 78% (7/9) in the conventional surgery group ( χ2=7.929, P=0.019). Comparisons between the postoperative CT and preoperative simulation CT images showed that the position deviation of the resected bones was (0.21 ± 0.19) mm. Conclusions:In the present study, improved precision of zygomatic bone resection and bone setback was achieved in reduction malarplasty by using the angled double L-shaped osteotomies with computer assistance. Moreover, complication occurrences (asymmetry, bone nonunion etc.) were significantly decreased. Also, patients’ expectation was better achieved with this method.

BACKGROUNDRoad traffic injuries (RTIs) are a worldwide issue associated with increasing development and motorization. However, statistical studies do not include any analyses of Beijing's geriatric population. Using data from the Beijing Emergency Medical Center, we present the main characteristics of traffic injuries involving the elderly in Beijing. We also provide objective information for those concerned with the safety of traffic systems and the prevention of traffic injuries.

METHODSIn a longitudinal, retrospective study, data were collected on 1706 victims aged 65 years and older who sustained traffic injuries in Beijing between 2004 and 2010. Personal information, time of injury event, emergency care response time, road user type, striking vehicle type, injury site, and severity of injury were analyzed using χ(2) tests and Logistic regression analysis.

RESULTSThe annual rate of traffic injuries was 21.80 per 100 000 elderly people in Beijing, and the morbidity rate decreased from 2004 to 2010 (P < 0.001). The mean age was (72.92 ± 5.67) years, and 911 (53.40%) of the victims were male. The majority of victims sustained head and lower limb injuries and were classified as being of medium severity. Traffic collisions occurred most frequently in the daytime excluding rush hours; these collisions included being hit by a car (85.64%) and pedestrian victim injuries (79.19%). Our statistical analysis found three factors for injury severity: abdominal injuries (P < 0.001), number of injury sites (P = 0.027), and head injuries (P = 0.034). The decline in traffic injuries is due to a decrease in victims aged 65-74 years and pedestrians; the severity of RTIs also decreased.

CONCLUSIONSThis study highlights the declining trend in traffic injuries among older adults in Beijing. However, traffic injuries remain a serious public health problem for the elderly and effective measures are required to reduce their incidence.

Accidents, Traffic ; statistics & numerical data ; Aged ; China ; epidemiology ; Emergency Medical Services ; Female ; Humans ; Male ; Wounds and Injuries ; epidemiology

BACKGROUNDSeveral isoforms of p53 have been reported, which may have varying functions and expressions. This study aimed to analyze the expression patterns of p53 isoforms in renal cell carcinoma (RCC) at the mRNA and protein levels and their associations with clinical and pathologic factors to explore the mechanism of p53 isoforms' activity in RCC.

METHODSThe specimens of tumours (T) and clinically normal tissues (N) adjacent to them were collected from 41 patients with RCC. mRNA expression levels of p53 isoforms were detected using RT-PCR followed by nested PCR. Protein expression levels were detected using immunohistochemisty and Western blotting with the anti-p53 antibodies DO-1 and DO-12. The data were analyzed with clinicopathological features by chi(2) test or Fisher's exact test.

RESULTSp53 mRNA was expressed in all tumours and matched clinically normal tissue adjacent to the tumour. All six isoforms could be detected in tumour and normal tissues, with the exception of the Delta133p53beta isoform, which was not detected in the normal tissue. Of the six isoforms, p53beta mRNA was significantly overexpressed in tumour samples (P < 0.001), and correlated with tumour stage. Nested PCR results consistently indicated the presence of p53gamma (19T/22N), Delta133p53 (33T/26N), Delta133p53beta (2T/0N), and Delta133p53gamma (13T/9N). Immunohistochemical analysis showed that p53 was expressed only in tumour tissues and correlated with tumour stage and grade. The results of Western blotting analysis were consistent with these findings.

CONCLUSIONSAlthough all six isoforms are present in RCC, their function in tumour development or progression might be different. Our findings suggest that p53beta might play an important role in the formation of RCC and it might be used as a new predictor and therapeutic target for RCC.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Carcinoma, Renal Cell ; genetics ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; physiology ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Protein Isoforms ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Young Adult

OBJECTIVETo study the clinical, pathologic and radiologic features of chondroblastoma occurring in sites other than epiphysis and apophysis of long bones, and to investigate possible reasons for misdiagnosis.

METHODSThe clinical, pathologic and radiologic data of 18 chondroblastoma cases occurring in atypical sites were collected from 5 major hospitals in Shanghai during the past 12 years. S-100 immunostaining was performed to confirm the cartilaginous differentiation of the tumor cells.

RESULTSChondroblastoma occurred in small bones of feet in 10 of the 18 cases (55.6%) studied, being commonest in the talus and calcaneus bones. Mean age of the patients was 27.8 years, with 55.6% over 25 years of age. Radiologic examination revealed expansive, multilocular and well-demarcated radiolucent lesions in most cases. There was local cortical destruction in 5 cases (28%) and soft tissue infiltration in 1 case. In 10 cases (55.6%), the tumor was associated with aneurismal bone cyst or simple bone cyst formation. None of the cases studied was accurately diagnosed clinically before the operation. In 2 cases, the pathology was also misdiagnosed, often being diagnosed as aneurismal bone cyst or giant cell tumor.

CONCLUSIONSChondroblastoma occurring in atypical sites are often associated with atypical age, radiologic features and pathologic findings at presentation. Thorough understanding of the potential pitfalls is essential in order to avoid misdiagnosis.

Adolescent ; Adult ; Bone Cysts, Aneurysmal ; diagnosis ; diagnostic imaging ; pathology ; Bone Diseases ; diagnosis ; diagnostic imaging ; pathology ; Bone Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Calcaneus ; Child ; Chondroblastoma ; diagnosis ; diagnostic imaging ; pathology ; Diagnostic Errors ; Female ; Giant Cell Tumor of Bone ; diagnostic imaging ; pathology ; Humans ; Male ; Radiography ; Talus

OBJECTIVESTo study on the changes of intracellular calcium and magnesium in cirrhosis and its clinical significance.

METHODSThe calcium and magnesium were determined in serum (SCa, SMg), platelets (PCa, PMg), mononuclear cells (MNCCa, MNCMg), polymorphonuclear cells (PMNCa, PMNMg) and erythrocytes (RCa, RMg) of 50 patients with uncompensative cirrhosis (group A) and of 35 patients with compensative cirrhosis (group B). 35 health persons were the control group.

RESULTSThe SCa and SMg of group A were lower significantly than those of both group B and control group. The MNCCa, PMNCa, RCa, PMg, MNCMg, PMNMg, RMg of group A [(4.76+/-1.91) micromol/10(9), (7.56+/-2.88) micromol/10(9), (0.66+/-0.13) mmol/L, (5.53+/-2.25) micromol/10(11), (6.64+/-3.53) micromol/10(9), (10.12+/-4.32) micromol/10(9), (2.02+/-0.76) mmol/L] and those of group B [(5.34+/-2.41) micromol/10(9), (8.32+/-2.34) micromol/10(9), (0.67+/-0.11) mmol/L, (5.55+/-2.67) micromol/10(11), (6.56+/-3.44) micromol/10(9), (10.95+/-4.45) micromol/10(9), (2.21+/-0.74) mmol/L] were lower significantly than those of control group [(6.86+/-2.02) micromol/10(9), (9.89+/-3.23) micromol/10(9), (0.72+/-0.10) mmol/L, (7.43+/-2.78) micromol/10(11), (8.68+/-4.1) micromol/10(9), (13.96+/-5.76) micromol/10(9), (2.74+/-0.92) mmol/L]; t (group A vs. control group)=4.88, 3.48, 2.31, 3.45, 2.46, 3.52, 4.00, 0.01, 0.01, 0.05, 0.01, 0.02, 0.01, 0.01; t (group B vs. control group)=2.87, 2.34, 2.00, 2.89, 2.33, 2.45, 2.65, 0.01, 0.05, 0.05, 0.01, 0.05, 0.02, 0.02. The PCa of the patients with hepatic encephalopathy was higher, the SMg, PMg, MNCMg, PMNMg and RMg were lower than those of the patients without hepatic encephalopathy significantly. The SCa, SMg, PMg, MNCMg, PMNMg and RMg of the patients in Child stage C were lower significantly than those of the patients in Child stage B. There were no significant differences of PCa, MNCCa, PMNCa and RCa between Child stage C and Child stage B. There were no significant differences of SCa, MNCCa, PMNCa and RCa between the patients with and without hepatic encephalopathy. The ratios of PCa/SCa, MNCCa/SCa and PMNCa/SCa of the patients with decreased SMg were lower than those of control group. The SMg, MNCMg, PMNMg and RMg were correlated directly with the level of serum albumin.

CONCLUSIONThere are calcium and magnesium deficiencies in the patients with uncompensative cirrhosis and compensative cirrhosis, this deficiency aggravates with the severity of the disease. There is relative increase of intracellular calcium. The magnesium deficiency may be one of the reasons for both hepatic encephalopathy and relative increase of intracellular calcium.

Adult ; Blood Cells ; chemistry ; Calcium ; blood ; Female ; Humans ; Liver Cirrhosis ; blood ; Magnesium ; blood ; Male ; Middle Aged

OBJECTIVETo determine whether vitamin D receptor(VDR) gene start codon polymorphisms and 3'-end region polymorphisms exerted a combined influence on bone mineral density(BMD) in Han postmenopausal women in Beijing area.

METHODSThe VDR Fok I and 3'-end region genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism in 110 unrelated postmenopausal women. BMD was measured at the lumbar spine (L(2-4)), femoral neck(Neck), Ward's triangle(Ward's) and trochanter (Troch) using duel-energy X-ray absorptiometry.

RESULTSThe frequencies distribution of Fok I, Apa I, Bsm I and Taq I alleles in this cohort all followed the Hardy-Weinberg equilibrium. No significant association of Fok I, Apa I or Taq I genotype with BMD in postmenopausal women was found when these polymorphisms were considered independently, except for Bsm I genotype. When a combined analysis of VDR gene Fok I and 3'-end region polymorphisms was carried out, cross-genotyping Fok I and Apa I polymorphisms was significantly associated with BMD at the L(2-4) (P<0.001), and cross-genotype of Fok I and Taq I was also significantly associated with BMD at the Neck and Troch sites (P<0.05). However, cross-genotyping Fok I and Bsm I polymorphisms was not significantly associated with BMD. Cross-genotyping Apa I and Bsm I or Taq I polymorphisms was not associated with BMD in postmenopausal women, either.

CONCLUSIONAlthough Fok I polymorphisms of VDR gene were not significantly associated with BMD in postmenopausal women, VDR gene Fok I and 3'-region polymorphisms (Apa I and Taq I) had a combined effect on the BMD in postmenopausal women.

3' Flanking Region ; genetics ; Aged ; Analysis of Variance ; Bone Density ; physiology ; China ; Codon, Initiator ; genetics ; DNA ; genetics ; metabolism ; DNA Restriction Enzymes ; metabolism ; Female ; Gene Frequency ; Genotype ; Humans ; Middle Aged ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Postmenopause ; genetics ; physiology ; Receptors, Calcitriol ; genetics