Objective:To clone the hTERT gene promoter and detect its transcriptional activities in various hepatocellular carcinoma cell lines and in human fibroblast cells in order to make groundwork for the targeting gene therapy for human hepatocellular carcinoma.Methods:The fragment of hTERT promoters were acquired by PCR amplification and were cloned into the reporter plasmid pGL3-Basic which contains a luciferase gene.The constructed eukaryotic expression plasmid pGL3-hTERT in which the expression of the luciferase were drived by hTERT promoter were transfected into three HCC cell lines and the primary fibroblast cells.At 24 hours post transfection (p.t.),the activity of the luciferase was determined with Dual-Luciferase Reporter Assay System.A pGL3-AFP containing luciferase gene controlled by the AFP promoter was used as positive control and a pGL3-Basic vector which have no promoter in front of the luciferase gene was used as negative control.Results:The hTERT promoter was successfully cloned into the eukaryotic reporter vector,pGL3-Basic and gained the pGL3-hTERT vector.The hTERT promoters showed high transcriptional activities in all the three HCC cell lines but no transcriptional activities in the primary fibroblast cells.The transcriptional activity of the hTERT promoter was much higher than AFP promoter(14～30 times).Conclusion:Our data revealed that the hTERT promoter possesses the tumor-spesific high transcrioptional activity in all the three established HCC cell lines.It may serve as a useful tool for transcriptional targeting of hepatocellular carcinoma gene therapy.

Objective To explore the clinical curative effect of traditional Chinese medicine enema,amino salicylic acid preparation and probiotics in the treatment of ulcerative colitis.Methods 168 patients were selected as the research objects which were confirmed by endoscopic examination and the diagnosis of ulcerative colitis patients,randomly divided into the control group of 84 cases and observation group of 84 cases.The control group was given with sulfasalazine(SASP) treatment,a dose of 1g,4 times/d,period of treatment was 28d.In the observation group,oral SASP treatment combined with microecological preparation often meian and traditional Chinese medicine enema,Chang An capsule,2 capsules (500mg)/time,3 times/d,postprandial oral; Herbal Enema with angelica,white lentils,flavescent sophora root,cicada,windproof suffering after filtering,every night before bed retention enema.The clinical efficacy was compared between the two groups.Results After the treatment,the Sutherland index of the observation group was (1.82 ± 1.65),which was significantly lower than (5.51 ± 1.59) of the control group (t =10.436,P ＜ 0.05).In observation group,the clinical total effective rate,the total efficiency of endoscopic and histological total effective rate was 88.1％,81％,76.2％,the control group were 66.7％,57.1％,50％,the two groups had significant difference(x2 =4.353,4.512,5.113,P＜0.05).Conclusion The traditional Chinese medicine enema,amino salicylic acid preparation and probiotics in the treatment of ulcerative colitis has good curative effect,can relieve the clinical symptoms of patients,improve the degree of intestinal diseases.

[Objective]To investigate the effect of using Minimully invasive percutuneous plate osteosunthosis to insert DHS for fixation of intertrochanteric hip fractures in elderly patients,in order to reduce operative trauma.[Method]To study retrospectivly 75 cases of the intertrochanteric fracture of elderly patients treated with DHS,these patients were treated from july 2004.7 to october 2005.10.MIPPO was used in 29 out of 75 cases,classic technique for fixation was used in other ones.[Result]All patients successfully discharged except two died because of fungal septicemia and lung embolism.Totally 73 cases were followed up from 7 to 21 months(averagd 12.3 months),one died of morbus internus in follow-up period.All the fractures had successful union,union time in average was 3.5 months,the diference between MIPPO group and classic incision group was not distinguished.Blood loss and transfusion in MIPPO group was less than classic incision group.Recovery of ESR and detumescence of the femoral was quickly in minimal incision group,time in bed was short.[Conclusion]Technique of minimal invasive fixation using DHS has little trauma and quick functional recovery for patients,therefore,this technique should be better for treatment of intertrochanteric fracture in elderly patients.

Abstract: Objective　To analyze the incidence of pulmonary tuberculosis infection in the elderly(&ge; 65 years old) in Xianning City from 2011 to 2021, and to provide scientific basis for formulating strategies and measures for tuberculosis prevention and treatment in the elderly. Methods　Descriptive epidemiological analysis was used to analyze the epidemic characteristics and influencing factors of elderly (this article studies elderly people aged 65 and above) pulmonary tuberculosis in Xianning City, Hubei Province from 2011 to 2021. Results　From 2011 to 2021, the average annual registration rate of elderly patients with pulmonary tuberculosis in Xianning City, Hubei Province was 211.52/100 000 (5 289/2 500 496), which showed a downward trend （χ2 trend=58.06, P<0.05), and significantly higher than the average registration rate of 96.04/100 000 patients with pulmonary tuberculosis (26 463/27 554 195)(χ2=2 896.69, P<0.05). The average proportion of the registered number of elderly patients with pulmonary tuberculosis in the whole population was 19.99% (5 289/26 463), which showed an upward trend (χ2trend=272.05, P<0.05). The average positive rate of etiology in elderly patients with pulmonary tuberculosis was 51.86% (2 743/5 289), which was significantly higher than that in the whole population 46.41% (12 282/26 463) (χ2=52.53, P<0.05). The average proportion of retreated patients in elderly patients with pulmonary tuberculosis was 8.92% (472/5 289), which was significantly higher than that of retreated patients in the whole population 6.33% (1 674/26 463) (χ2=47.23, P<0.05). The sex ratio of male to female in elderly patients with pulmonary tuberculosis was 2.92∶1. Among the six counties (districts) under the jurisdiction of Xianning City, Hubei Province the registration rate of elderly tuberculosis patients in Xian'an district was the highest 262.20/100 000 (1 267/483 220), and that in Tongcheng County was the lowest 123.83/100 000 (575/464 338), with significant differences (χ2=233.67, P<0.05). Conclusion　The overall incidence of tuberculosis in the elderly in Xianning City , Hubei Province showed a downward trend, but the proportion of the incidence of tuberculosis in the elderly in the whole population has increased year by year, suggesting that the elderly are the key population of tuberculosis prevention and control in Xianning City in the future, and comprehensive prevention and control measures should be taken to reduce the incidence of tuberculosis in the elderly population.

Objective To compare the changes of some blood indices after treatment of intertro-chanteric fractures with conventional and minimally invasive dynamic hip scres (DHS) internal fixation so as to understand the influence of minimally invasive technique on physiology of the organism. Meth-ods The elderly patients with intertrochanteric fractures in our department from July 2004 to May 2006 were divided randomly into two groups, ie, conventional DHS internal fixation group(Conventional group, 52 patients)and the minimally invasive DHS internal fixation group(Minimal invasion group, 54 patients). A comparison was done on data including white blood cells (WBC), hemoglobin (Hb), e-rythrocyte sedimentation rate (ESR). C reactive protein (CRP) and creatine kinase (CK) as well as re-cessive blood loss. Results The indices including transfusion, blood loss, recessive blood loss, ESR and CK in minimal invasion group showed less changes compared with conventional group. With statistical difference. But WBC and CRP showed no statisfical difference between both groups. Conclusion Minimally invasive DHS internal fvtation carl reduce operative trauma during treatment of intertrochanterie fractures in the elderly.

Objective To study the protective effect of vitamin C on UV-induced photoaged skin structure in rats and provide a basis for clinical medicine and health care.Methods Photoaging skin models were set up by chronic ultraviolet radiation in 24 SPF female Sprague-Dawley rats, irradiated twice weekly for 4 weeks.The rat photoaged skin was induced by exposure to a total dose of 123 J/cm2 UVA and 5 J/cm2 UVB for 4 weeks.Vitamin C was administered by gastric gavage in a dose of 50 mg/kg once daily for 30 days during the model development.We compared the pathological changes in the irradiated skin using HE, Masson’ s trichrome and Victoria blue B staining, and compared the ultrastructural changes by electron microscopy.Results Rat models with photoaged skin were developed successfully.The vitamin C group showed significant reduction of pathological severity including erythema, ulcers, epidermal cell proliferation, epidermal thickness, dermis vasodilation, inflammatory cell infiltration, fibroblast proliferation, endoplasmic reticulum dilatation, mitochondrial swelling and vacuolization, elastic fibers degeneration and focal accumulation, collagen fibers swelling with uneven thickness,compared with the rats of model group at the irradiation site.Conclusions Vitamin C is effective in reducing the structural damage of UV-induced photoaged skin in rats.

OBJECTIVETo detect and compare the transcriptional activities of tumor-specific survivin and AFP promoters in various hepatocellular carcinoma cell lines and to lay some groundwork for targeting gene therapy in human hepatocellular carcinoma.

METHODSThe fragment of survivin and AFP promoters were acquired by PCR amplification and were cloned into the reporter plasmid pGL3-Basic, which contained a luciferase gene. The constructed eukaryotic expression plasmid pGL3-SUR and pGL3-AFP, in which the expression of the luciferase was derived by survivin or the AFP promoter, were transfected into three HCC cell lines. At 24 hours post transfection (p.t.), the activity of the luciferase was determined with Dual-Luciferase Reporter Assay System. A pGL3-CMV, containing the CMV promoter controlled luciferase gene, was used as a positive control.

RESULTSBoth survivin and AFP promoters had transcriptional activities in all three HCC cell lines and the transcriptional activity of the survivin promoter was much higher than the AFP promoter (52-98 times) and reached a level of 16% approximately 21% of the transcriptional activity of the CMV promoter.

CONCLUSIONOur data reveals that the survivin promoter possesses a high transcriptional activity in all three established HCC cell lines and may serve as a useful tool for transcriptional targeting gene therapy of HCCs.

Carcinoma, Hepatocellular ; genetics ; Gene Targeting ; Genetic Therapy ; Humans ; Inhibitor of Apoptosis Proteins ; Liver Neoplasms ; genetics ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; Promoter Regions, Genetic ; genetics ; Transcription, Genetic ; Transfection ; Tumor Cells, Cultured ; alpha-Fetoproteins ; genetics

Objective:To classify the symptom characteristics of young and middle-aged peritoneal dialysis patients and to explore the differences of demographic functional status and social function of patients with different symptom categories.Methods:A total of 179 peritoneal dialysis patients from 3 peritoneal dialysis centers in Shanghai were investigated from December 2019 to August 2020 by General Information Questionnaire, Peritoneal Dialysis Symptom Distress Scale. Latent class analysis was used to classify young and middle-aged peritoneal dialysis patients according to symptom characteristics. Multiple Logistic regression was used to explore the differences of demographic and disease characteristics of different categories of patients.Results:Peritoneal dialysis patients could be divided into three potential categories according to symptom characteristics ( P<0.05). According to the conditional probability of each category, they were named "low symptom group" (111 cases,62.0%), "high psychological-moderate physical symptom group" (22 cases, 12.3%), "high symptom group" (46 cases,25.7%). There were differences in working status ( OR=0.029, P<0.01), education level ( OR=152.799, P<0.01), duration ( OR=81.307, P<0.05), diabetic nephropathy ( OR=80.619, P<0.01) and CCI score ( OR=91.188, P<0.01) distribution among different potential categories of young and middle-aged peritoneal dialysis patients. Conclusions:There are three types of potential symptoms in young and middle-aged patients undergoing peritoneal dialysis. In clinical practice, medical staff should focus on the psychological status of young and middle-aged patients with low educational background and early stage of dialysis, and encourage them to return to work; at the same time, they should regularly evaluate the symptom burden of patients with diabetic nephropathy and high complication index peritoneal dialysis patients, in order to provide targeted intervention measures to prevent the progression of the disease.

Endometriosis is frequently associated with infertility. It is believed that the impairment of endometrial receptivity may be one of the mechanisms involved in this condition. Endometriosis results in infertility by affecting the quality of the embryo and decreasing the endometrium tolerance.Among patients with endometriosis and infertility,a large number of clinical applications found that even if the embryo quality was good, the embryo's planting rate was still low, however, in the case of good endometrial receptivity,abnormal embryos have possible bed success.This paper mainly reviews the endometrium receptivity in patients with endometriosis and the correlation between the regulation factors of endometrial receptivity and infertility, with a view to improving the clinical pregnancy rate.

OBJECTIVESTo explore the feasibility of DC being in vitro induced from AML cells with cytokine cocktails and their biological properties.

METHODSAML cells were cultured in either presence or absence of cytokine cocktails. DC were studied for morphology, and cytochemical and immunofluorescent staining. Functions of DC were examined by MLC, FITC-conjugated dextran uptake test, and LDH release assay. RT-PCR and FISH were used to analyze the specific fusion genes of culture-derived DC.

RESULTSClassical DC morphological changes occurred in all 15 cultured AML cells. DC-associated surface molecules such as CD(1a), CD(80), CD(86), CD(106), CD(83) and HLA-DR were upregulated (P < 0.05). The allostimulatory abilities of culture-derived DC were significantly higher than those of AML cells uncultured or cultured in the absence of cytokines (P < 0.05). Culture-derived DC only in the presence of GM-CSF + IL-4 have phagocytotic activities. CTL assay was performed in 5 of the 15 samples. At effector/target ratio of 20:1, auto-T lymphocytes primed with the culture-derived DC exhibited no more killing activity to auto-AML cells than those stimulated by IL-2 or uncultured AML cells. Culture-derived DC presenced the native AML-specific aberrant karyotype and related fusion gene.

CONCLUSIONSCytokine cocktails could in vitro induce AML cells into DC with classical morphology, immunophenotype and function. DC maturity induced by different cytokine cocktails could be variable. Culture-derived DC were originated from the native AML cells. AML cells could make the auto-T lymphocyte anergy.

Adolescent ; Adult ; Aged ; Cell Differentiation ; drug effects ; Cytokines ; pharmacology ; Dendritic Cells ; cytology ; Drug Synergism ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; In Vitro Techniques ; Interleukin-4 ; pharmacology ; Leukemia, Myeloid, Acute ; immunology ; pathology ; Male ; Middle Aged ; Monocytes ; cytology ; drug effects ; Tumor Necrosis Factor-alpha ; pharmacology