ObjectiveTo investigate the effect of heat treatment combined with narrow band ultraviolet B(NB-UVB) on cultured normal human melanocytes in vitro.MethodsMelanocytes were isolated from the foreskin of normal human,cullured in vitro,and irradiated with NB-UVB of different doses(20,30,50,70,90,120 and 180 mJ/cm2).Then,MTT assay was performed to evaluate the proliferation and activity of melanocytes to determine the optimal dose of UVB for the next experiment.Melanocytes were classified into 3 groups to be treated with heat at 42 ℃ for 1 hour (heat group),irradiated with UVB at 50 mJ/cm2 (UVB group),or irradiated with UVB at 50 mJ/cm2 followed by heat treatment at 42 ℃ for 1 hour (combination group),daily for 3 successive days; those receiving no treatment served as the control.After 24-hour culture following the last treatment,tyrosinase activity was evaluated with L-dopa as the substrate,melanin content was detected by NaOH assay,and cell cycle stages were determined by flow cytometry.ResultsNB-UVB irradiation decreased the viability of melanocytes in a dose-dependent manner,and the optimum dose of UVB was 50 mJ/cm2.The tyrosinase activity of melanocytes was 0.244 ± 0.018 and 0.310 ± 0.015 respectively in the UVB group and combination group,and increased by 3.8％ (P ＜ 0.05) and 31.9％ (P ＜ 0.05) respectively compared with the control group (0.235 ± 0.018); the melanin content was 0.201 ± 0.016 and 0.286 ± 0.019,respectively in the UVB group and combination group,and increased by 17.5％ (P ＜ 0.05 ) and 67.3％ (P ＜ 0.05) compared with the control group (0.171 ± 0.016).In comparison with the control group,the percentage of melanocytes in G1 phase was decreased by 23.94％ in the UVB group(P＜ 0.05) and 33.51％ in the combination group(P ＜ 0.05),while that in S phase and G2 phase increased by 15.35％ (P ＜ 0.05 ) and 11.93％ (P ＜ 0.05),respectively in the UVB group,and 17.76％ (P ＞ 0.05) and 16.08％ (P ＞ 0.05),respectively in the heat group.ConclusionHeat treatment and NB-UVB can synergistically enhance the tyrosinase activity and accelerate melanogenesis,proliferation and differentiation,of melanocytes.

ObjectiveTo explore the effects of heat treatment and ultraviolet B (UVB) radiation alone or in combination on the expression of heat shock protein (HSP) 72 in human epidermal melanocytes.Methods Melanocytes were obtained from human foreskin,and subjected to primary culture.After 3 to 5 passages,the melanocytes were classified into 4 groups:control group (receiving no treatment),heat treatment group (treated with heat at 42 ℃ for 1 hour every day for 3 days),UVB group(irradiated with UVB at 50 mJ/cm2 daily for 3days),combination group(treated with heat at 42 ℃ for 1 hour followed by irradiation with UVB at 50 mJ/cm2daily for 3 days).After another 2- to 6-hour culture following the last treatment,melanocytes were collected and subjected to real time PCR and Western blot for the detection of HSP72 mRNA and protein expression,respectively.ResultsThe mRNA and protein expressions of HSP72 were significantly higher in the heat treatment group and combination group than in the control group (mRNA:6.584 ± 0.871 and 7.269 ± 0.454 vs.0.975 ± 0.089,both P ＜ 0.001; protein:2.022 ± 0.058 and 2.080 ± 0.045 vs.0.532 ± 0.033,both P ＜ 0.001 ),but was similar between the UVB group and control group (mRNA:0.832 ± 0.084 vs.0.975 ± 0.089,P ＞ 0.05;protein:0.546±0.021 vs.0.532 ± 0.033,P ＞ 0.05).The ANOVA of factorial design showed that neither heat treatment nor UVB irradiation had interaction effect on the mRNA or protein expression of HSP72 (F =2.106,1.399 respectively,both P ＜ 0.05).ConclusionsHeat treatment can cause an increase in the expression of HSP72,which may enhance the function of melanocytes and protect melanocytes from UVB induced damage.

Medical education should combine clinical professional skills with humanities skills,integrate humanities knowledge into vocational education.Taking 2008 grade seven-year program clinical medicine students in the 2nd affiliated hospital of Harbin Medical University as fostering object,we made researches into students' understanding of doctor-patient relationship before practice and their mastering of skills.Through conducting questionnaire,we got to know the effect of humanities practice skill training for seven-year program clinical medicine students.Meanwhile,we compared students' self evaluation results before and after training,discussed on how to improve medical students' communication skills,cultural skills and the reform direction in an aim to guide students to transit from students to clinical doctor.

ObjectiveTo observe the increasing effect of infrared irradiation on tyrosinase activity and melanin content in cultured normal human epidermal melanocytes in vitro and to explore the optimal dose of infrared irradiation.MethodsEpidermal melanocytes were isolated from normal human foreskin tissue,and subjected to primary culture.Methyl thiazolyl tetrazolium(MTT) assay was performed to evaluate the effect of different doses(0,20,60,80,100,140,240,320 J/cm2)of infrared light on the proliferation of melanocytes and to select the optimal irradiation dose.Then,melanocytes were irradiated with infrared light at the optimal dose for 3 consecutive days followed by the determination of tyrosinase activity,melanin content,and cell cycle via dopa oxidation assay,NaOH solubilization method and flow cytometry,respectively.ResultsThe best intervention dose of infrared light was 80 J/cm2.The tyrosinase activity(A492 nm) and melanin content(A492 nm)were 0.3601 ± 0.0301 and 0.2748 ± 0.0243 respectively in melanocytes after irradiation with infrared light of 80 J/cm2 for 3 days,significantly higher than those in unirradiated melanocytes(0.3114 ± 0.0341,0.2325 ±0.0254,respectively,both P ＜ 0.05),with an increase rate of 15.64％ and 18.19％ respectively.Cell cycle analysis revealed a decline in cell percentage in G1 phase(P ＜ 0.01 ) but a concomitant increase in cell percentage in G2 and S phase (both P ＜ 0.05) in irradiated melanocytes compared with unirradiated melanocytes.ConclusionsThe optimal dose of infrared light is 80 J/cm2 for the irradiation of melanocytes,and this dose of infrared light can increase melanin content,tyrosinase activity,differentiation and proliferation of melanocytes.

BACKGROUNDThe vitamin D receptor (VDR) gene has been identified as a candidate gene for susceptibility to Parkinson's disease (PD), but results from genetic association studies to date are inconsistent. Here, we conducted a meta-analysis of published case-control studies to evaluate the association of the extensively studied VDR ApaI (G/T), BsmI (G/A), FokI (C/T), and TaqI (T/C) gene polymorphisms with risk of PD.

METHODSElectronic search at PubMed, EMBASE, EBSCO, China National Knowledge Infrastructure, Weipu database, and Wanfang database was conducted to identify all relevant studies. Odds ratio (OR) with 95% confidence interval (CI) values was applied to evaluate the strength of the association.

RESULTSA total of seven studies with 2034 PD cases and 2432 controls were included in the meta-analysis following the inclusion and exclusion criteria. Overall, no significant association between ApaI, BsmI, and TaqI gene polymorphisms and PD susceptibility in all four genetic models was found (T vs. G: OR = 1.00, 95% CI: 0.89-1.12, P = 0.97; A vs. G: OR = 0.94, 95% CI: 0.77-1.15, P = 0.53; C vs. T: OR = 1.03, 95% CI: 0.85-1.25, P = 0.77) while a significant association between FokI (C/T) and PD risk was observed (C vs. T: OR = 1.41, 95% CI: 1.14-1.75, P = 0.001; CC vs. TT: OR = 2.45, 95% CI: 1.52-3.93, P = 0.0002; CT vs. TT: OR = 2.21, 95% CI: 1.38-3.52, P = 0.0009, CC vs. CT+TT: OR = 2.32, 95% CI: 1.49-3.61, P = 0.0002).

CONCLUSIONSPolymorphisms of ApaI, BsmI, and TaqI may not be associated with the susceptibility to PD while the FokI (C/T) polymorphism is possibly associated with increased PD risk. However, conclusions should be cautiously interpreted due to the relatively small number of studies included.

Genetic Predisposition to Disease ; genetics ; Humans ; Parkinson Disease ; genetics ; Polymorphism, Genetic ; genetics ; Receptors, Calcitriol ; genetics

OBJECTIVETo study the clinical manifestations, radiologic findings, pathologic diagnosis and differential diagnosis of primary osteosarcoma in elderly patients.

METHODSTwelve cases of primary osteosarcoma occurring in patients older than 60 years were encountered during the period from 1985 to 2010. The clinical manifestations, radiologic features and pathologic findings were studied and the follow-up data were analyzed.

RESULTSThe sites of involvement included long bones (number = 7), ilium (number = 1), craniofacial bones (number = 2) and soft tissue (number = 2). Radiologic examination showed a mixture of osteosclerotic and osteolytic lesions in 10 patients, soft tissue lesions with high-density areas in 2 patients and soft tissue lesions with periosteal reaction in 8 patients. Histologically, most cases showed features of conventional osteosarcoma. There were 2 cases of malignant fibrous histiocytoma-like osteosarcoma, 2 cases of chondroblastic osteosarcoma and 1 case of well-differentiated intraosseous osteosarcoma. Immunohistochemical study played little role in pathologic diagnosis. Ten patients had undergone amputation, including one patient who had received adjuvant chemotherapy beforehand. Nine patients had follow-up information available. Three of them died of lung metastasis and 1 died of cardiovascular disease.

CONCLUSIONSPrimary osteosarcoma rarely occurs in elderly patients and can easily be missed. Correlation with clinical, radiologic and histologic features is important for arriving at a correct diagnosis.

12E7 Antigen ; Aged ; Antigens, CD ; metabolism ; Bone Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Cell Adhesion Molecules ; metabolism ; Chondrosarcoma ; pathology ; Diagnosis, Differential ; Female ; Femoral Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Humans ; Ilium ; Lung Neoplasms ; secondary ; Lymphoma ; pathology ; Male ; Middle Aged ; Osteitis Deformans ; pathology ; Osteosarcoma ; diagnostic imaging ; metabolism ; pathology ; surgery ; Radiography ; Soft Tissue Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

OBJECTIVETo investigate the clinical features of unexpected sudden death (SUD) clustered in families in Yunnan province.

METHODSThis retrospective study analyzed the clinical features of SUD occurred between July to September 2005 in 7 families in Yunnan province.

RESULTSAll 16 SUD patients shared common clinical features such as fatigue and repeated syncope and one group of SUD patients (n = 8 from 4 families) presented with the gastric intestinal tract manifestations including nausea, vomiting, abdominal pain and diarrhea with suspected dietary history and abnormal laboratory enzyme findings (GOT/GPT, CK/CKMB, LDH/LDH1 etc.). In SUD patients without gastric intestinal tract manifestations (n = 8 from 3 families), there were no clear symptoms before death and repeated ventricular tachycardia and ventricular fibrillation were recorded in one survivor. There was no clear evidence for the involvements of hereditary and infectious factors for observed SUD.

CONCLUSIONThe reason for the unexpected sudden death clustered in 7 families in Yunnan remains unclear. Repeated syncope and fatigue served as the common clinical features in the presence or absence of gastric intestinal tract manifestations in all SUD cases. Further studies are needed to clarify the pathology and detailed clinical manifestations of SUD occurred in this area.

Adolescent ; Adult ; Bias ; Cause of Death ; Child ; China ; epidemiology ; Death, Sudden ; epidemiology ; Family ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Objective:To evaluate the clinical efficacy and safety of modified chitosan eye drops on rabbit Candida albicans keratitis model.Methods:Ten healthy female New Zealand rabbits were used to establish the superficial Candida albicans keratitis model by the corneal surface lens method in the right eye.Slit lamp microscopy and corneal scraping and microscopic examination were performed to preliminarily determine whether the keratitis model has been successfully established, the rabbits were then randomly divided into a model group and a modified chitosan group by the random number table method.The successfully established rabbit models which were determined by fungal culture results were retained.Five normal rabbits receiving no intervention served as a normal control group.The experimental eyes in the normal control and modified chitosan groups were treated with modified chitosan eye drops, Six times a day for one week, and subsequently four times a day for one week.No treatment was administered to the model group.The changes of corneal lesions and ocular surfaces were examined by slit lamp microscopy every day.At 1, 7, 14, 21 and 28 days after modeling, the eye condition and corneal clinical scores was assessed by slit lamp microscopy.The corneal conditions in each group was observed for two weeks after drug withdrawal.Results:The corneal scraping and microscopic examination results of eight rabbits models showed that the fungal hyphae and spores were positive.The fungal culture results showed that the separated pathogen was Candida albicans.The success rate of modeling was 80%(8/10). The clinical scores in the model group at 7, 14 and 21 days after modeling were 14.50±0.58, 6.25±0.50 and 2.50±0.58, respectively, and were significantly higher than 7.25±1.26, 2.75±0.50 and 1.25±0.50 in the modified chitosan group (all at P<0.05). In the model group, corneal edema was significantly aggravated, and the central white ulcer area was enlarged within seven days after modeling.Between 7 and 28 days after modeling, the corneal ulcer was gradually healed, while the central corneal scar and neovascularization were remained.The average healing time was (24.5±2.6)days.In the modified chitosan group, the corneal infiltration was significantly alleviated within seven days after modeling, and the fungal hyphae and spores of corneal scraping were negative on the 14th day after modeling.The average healing time in the modified chitosan group was (13.5±1.3)days, which was significantly shorter than that in the the model group (P<0.01). No recurrence of keratitis was observed in the modified chitosan group after two weeks of drug withdrawal.The cure rate was 100%.In the normal control group, the conjunctival hyperemia, corneal edema, and lesions were not observed during topical administration.Conclusion:The treatment with modified chitosan eye drop is effective in a rabbit superficial Candida albicans keratitis model, and have no obvious toxic effects on ocular tissues.

This study was purpose to investigate the expression of death-associated protein kinase (DAPK) gene in acute leukemia (AL) patients and the methylation status of its promoter region through experiments of DAPK methylation and expression, and to analyze the relation between them. The expression of DAPK gene in leukemia cells and normal bone marrow cells was detected by RT-PCR; the methylation status of DAPK gene promoter region in cells from AL patients and leukemia cell lines HL-60 and U937 was detected by nested methylation specific PCR (n-MSP); 2 randomly primers selected from randomly amplified products of second round nMS-PCR were cloned and sequenced in professional company. The results showed that the DAPK gene expressed in bone marrow specimens of all 10 normal controls, with average value of expression 0.92 ± 0.18, while the average value of DAPK expression in bone marrow specimens of AL patients was 0.61 ± 0.40 which was lower than that in normal controls (P < 0.05). The low or deletion of DAPK mRNA expression were found in bone marrow specimens of 9/17 (52.94%) cases of ALL and 42/102 (41.18%) cases of AML. The cell line U937 showed normal expression of DAPK gene, while cell line HL-60 showed the expression detection of DAPK gene. The methylation of DAPK promoter region existed in 33 out of bone marrow specimens of 102 AML patients and in 8 out of bone marrow specimens of 17 ALL patients, the methylation rates were 32.4% (33/102) and 47% respectively. The DAPK promoter region in bone marrow of 7 normal controls was unmethylated, while DAPK promoter region in U937 cells and HL-60 cells were unmethylated and methylated respectively. The DAPK mRNA expression in ALL and AML patients significantly negatively correlated with the methylation of its promoter region (r = -0.855, P < 0.05, in AML patients and r = -0.343, P < 0.05, in AML patients) suggesting the close relationship between them. It is concluded that the methylation of DAPK gene promoter region relates with abnormal expression or detection of DAPK mRNA in AL patients.
Acute Disease ; Case-Control Studies ; DNA Methylation ; Death-Associated Protein Kinases ; genetics ; HL-60 Cells ; Humans ; Leukemia ; genetics ; Promoter Regions, Genetic ; RNA, Messenger ; genetics

Objective To explore the efficacy and safety of recombinant human anti-severe acute respiratory syn-drome coronavirus 2(anti-SARS-CoV-2)monoclonal antibody injection(F61 injection)in the treatment of patients with coronavirus disease 2019(COVID-19)combined with renal damage.Methods Patients with COVID-19 and renal damage who visited the PLA General Hospital from January to February 2023 were selected.Subjects were randomly divided into two groups.Control group was treated with conventional anti-COVID-19 therapy,while trial group was treated with conventional anti-COVID-19 therapy combined with F61 injection.A 15-day follow-up was conducted after drug administration.Clinical symptoms,laboratory tests,electrocardiogram,and chest CT of pa-tients were performed to analyze the efficacy and safety of F61 injection.Results Twelve subjects(7 in trial group and 5 in control group)were included in study.Neither group had any clinical progression or death cases.The ave-rage time for negative conversion of nucleic acid of SARS-CoV-2 in control group and trial group were 3.2 days and 1.57 days(P=0.046),respectively.The scores of COVID-19 related target symptom in the trial group on the 3rd and 5th day after medication were both lower than those of the control group(both P＜0.05).According to the clinical staging and World Health Organization 10-point graded disease progression scale,both groups of subjects improved but didn't show statistical differences(P＞0.05).For safety,trial group didn't present any infusion-re-lated adverse event.Subjects in both groups demonstrated varying degrees of elevated blood glucose,elevated urine glucose,elevated urobilinogen,positive urine casts,and cardiac arrhythmia,but the differences were not statistica-lly significant(all P＞0.05).Conclusion F61 injection has initially demonstrated safety and clinical benefit in trea-ting patients with COVID-19 combined with renal damage.As the domestically produced drug,it has good clinical accessibility and may provide more options for clinical practice.