Objective To observe the effects of different dosages of granulocyte-macrophage colony-stimulating factor (GM-CSF) on generating the routine bone marrow dendritic cells, and supply suitable dosage of GM-CSF on preparation of dendritic cell vaccines used for different purpose. Methods Using low (5 ng/ml) and conventional (20 ng/ml) and high dosage( 50 ng/ml ) of GM-CSF combined interleukin-4 ( IL-4 ) to induce murine bone marrow dendritic cells were performed, The phenotypes (CD_(11c), CD_(80), CD_(86)) and functional properties of the DC were compared by FACS analysis and MLR. Results The DC induced by low dosage of GM-CSF were immature DC, expressing low CD_(11c), CD_(80), and CD_(86). DC induced by conventional dosage were functional mature, expressing higher CD_(11c), CD_(80), CD_(86), which could induce allogeneic T lymphocyte responses. DC induced by the high dosage GM-CSF were the most phetotypicaUy and functional mature cells, expressing the highest CD_(11c), CD_(80) CD_(86), which could induce the strongest allogeneic T lymphocyte responses. Conclusion The dosages of GM-CSF affect the maturation stage of dendritic cells. Low dosage of GM-CSF generated immature dendritic cells, but conventional dosage and high dosage generated mature dendritic cells. DC generated through high dosage of GM-CSF were the most mature in phenotype and function.

BACKGROUND:Foreign scholars have obtained a success to cure fracture by implanting the complex of red bone marrow and formation factor.Due to the in vitro culture process is not necessary,the complex of red bona marrow and scaffold formation factor is only required to be implant immediately,called uncellular tissue engineered bone.OBJECTIVE:This study innovatively constructs uncellular tissue engineered bone with autologous red bone marrow wrapped by fascial flap with pedicle,and validates the superiority of repairing bone defects.DESIGN,TIME AND SETTING:Homebody controlled animal experiment was performed in the Hebei North University and the Experiment Center of the Affiliated Hospital to Hebei North University from December 2007 to February 2009.MATERIALS:A total of 24 News Zealand albino rabbits,aged 4-5 months; uncellular tissue engineered bone was a mixture of autologous red bone marrow and osteoinductive absorbing materials containing bone morphogenetic proteins.METHODS:Bone defect models were induced on adult New Zealand rabbits' right radial bone,left side served as control group,only implanted with osteoinductive absorbing materials complex,while right side served as experiment group,which contained fascial flap with pedicle.A fascial flap prepared with capillary network containing nameless blood vessel pedicle was located to be adjacent to the bone defect using micro-surgical technique,to wrap the tissue engineered bone and to fill the bone defect.MAIN OUTCOME MEASURES:At 4,8,12,16 weeks postoperation,six rabbits were tested by radiograph,spectrodensitometry,gross morphology observation,histological inspection,quantitative analysis of bone morphometry in bone defect area and analysis of vessels image in the junctional zone.RESULTS:①X-ray determination:At 16 weeks,the implant surrounding bone defects formed bone shaft structure in the control group,cortical bone was not continuous and medullary cavity was obstructed; in the experiment group,normal bone shaft structure was formed and recanalization of medullary cavity was observed.②Histological observation:At 16 weeks,few vessels grew into implant in the control group,mature bone trabecular were observed,and medullary cavity was obstructed; in the experiment group,the implanting materials were completely degraded and substituted by new bone,mature bone structure formed and recanalization of medullary cavity was observed.③Quantitative analysis of bone morphometry in bone defect area:At 4,8,12,16 weeks postoperation,the volume of bone trabecula in the experiment group was more than that in control group (P ＜ 0.05).④Analysis of vessels image in the junctional zone:The area of vessels in the unit area in the experiment group was greater than that in control group in every time stage (P＜0.05).CONCLUSION:The uncellular tissue engineering complex constructed by autologous red bone marrow wrapped by fascial flap with pedicle shows double effects of hymeno-inducing regeneration of bone and the vascularization.It is feasible to repair large-segment bone defects.It has obvious therapeutic effect in the aspects such as reducing the bone defect reparation time and advancing the quantity and quality of the bone generation.

Objective To study the effect of fascial flap with vessels inducing the vascularization of uncel-lular tissue engingeering complex and the regenration of bone on the repair of bone defect, so as to provide the basis for the clinical application. Methods An animal model of bone defect on adult Newzland rabbits'right radial bone was established .and autologous red bone marrow were taken out and mixed into uncellulax tissue engineering comple-xes with OAM which contained BMP. The experiment animals were divided into two groups : experiment group and control group( n = 12 for each ). The control group was only implanted with complexes, meanwhile, the experiment group had fascial flap with vessels. By microsurgery technology,a non-named fascial flap with vessels was prepared, which belonged to capillary net,around the bone defect,and let it wrap tissue engineering complex,fill up bone de-fect. In a certian time, radiograph(X-ray) and light density measure was conducted, gross morphology and histological inspection was exmained. Bone shape measurement analysis and image of vessel analysis were conducted. All the sta-tistics were analyzed by the SPSS 11.5 software. Results Because of mechanically preventing fiber connective tis-sues and surrounding soft tissues from entering the areas of bone defect by fascial flap, it can keep bone defect having a relative stable environment ;The subfascial space itself, and also the shape and mass of filled-in subject had the de-cisive effect on the results of the regeneration of the bone; Owing to the establishment of blood supply during the con-structing tissue engineering complex. The experiment group was obviously superior to the control group. Compared with control group,the absor bance obviously increased in experiment group [(0. 732 ± 0. 021 ) vs (0. 651± 0.018)] (P < 0. 001 ) four weeks after the operation; also the bone trabecular body was significantly increased [(2.32±2.57)% vs(19.37±3.52)% ,(8.37±3.52)% vs(30.24±3.42)% ,(28.57±2.98)% vs(58.76± 4.62)% ,(47.24±3.42)% vs(88.72±5.84)%] ,and capillary area [(5.04±1.62)% vs(17.53±2.86)%, (10.37 ±2.96)% vs(35.24±1. 13)%,(18.20±2. 12)% vs(48.76±4. 62)%,(17.82 ±2. 74)% vs (57.72 ±5.84)%] (P <0.05) at each time period(4 weeks,8 weeks,12 weeks,and 16 weeks after operation). Despite of growth of implant's internal vessel, the number and speed of forming bone trabecula and cartilaginous tis-sue, even developing of mature bone structure, recreating of diaphysis structure, reconstructing of marrow cavity, ab-sorbing and decomposing of implant, the experiment group was obviously superior to the control group. Conclusions The induction of fascial flap with vessels shows double effects, one of which is the vascularization of uncellular tis-sue engineering complex and the other is membrane guided bone regeneration, So the method has a wonderful effect on the repair of bone defect.

Objective:To investigate clinical features, treatment and prognosis of herpes zoster during pregnancy or the perinatal period.Methods:Clinical data were collected from female inpatients with herpes zoster during pregnancy or the perinatal period in the First Affiliated Hospital of Zhengzhou University from January 2011 to December 2020, and clinical characteristics, treatment and prognosis were retrospectively analyzed.Results:A total of 25 patients were included in the study, including 22 pregnant patients at 10 - 38 gestational weeks (1 in the first trimester, 13 in the second trimester, and 8 in the third trimester) and 3 patients during the first postpartum week; their age ranged from 22 to 37 years, and the disease course varied from 2 to 9 days. Skin lesions were located on the head and face in 8 cases, on the chest and back in 5, on the waist and abdomen in 7, on the upper limbs in 1, on the lower limbs in 3, as well as on the perineum in 1; 3 patients presented with disseminated herpes zoster. The patients all presented with erythema, papulovesicles and blisters, which were accompanied by bloody bullae in 4 patients, by bullae in 3, as well as by erosions and exudations in 6; 5 patients had fever, 7 had mild pain, 6 had moderate pain, and 11 had severe pain. Sixteen pregnant patients and 3 postpartum patients received antiviral therapy (oral or intravenous infusion of acyclovir, vidarabine monophosphate, foscarnet sodium-sodium chloride injection) , 24 patients were treated with neurotrophic drugs, 1 pregnant patient and 3 postpartum patients received analgesic treatment, and other treatments included anti-infective therapy, wet compress, infrared radiation, etc. Preterm delivery occurred in 1 pregnant patient, other pregnant patients had full-term deliveries, and no abnormalities were observed in neonates. Sequelae occurred in 3 pregnant patients, including pain in 1 and pruritus in 2.Conclusion:Among the 25 patients, the treatment of herpes zoster during pregnancy or the perinatal period showed no obvious effect on their fetuses or newborns.

In order to study the tumor suppression effect of p53 with CMV enhancer and hTERT promoter mediated by human-derived vector pHrn in liver cancer cell Bel-7402, report plasmid pchEGFP, tumor suppressor plasmids pchp53Arg and pchp53Pro were constructed by inserting expression cassette CMVe+hTERTp+EGFP, CMVe+hTERTp+p53Arg and CMVe+hTERTp+p53Pro into pHrn respectively. 24 h after cell transfection by lipofectamine 2000, GFP expression pattern was analyzed through fluorescence microscope and flow cytometry; RT-PCR and Western blot were taken to study the p53 expression pattern. The cell apoptosis by Hoechst 33258 and Annexin V-FITC/PI staining was also studied. Results show that the expression of GFP and p53 protein in Bel-7402were detected, but apparent cell apoptosis could not be found. The recombinant p53 mediated by human-derived vector could express in Bel-7402, but no significant tumor suppression effect was detected, which might result from the down regulation effect of the wild type p53 on hTERT promoter.

Objective:To investigate differences in clinical characteristics between bullous pemphigoid (BP) patients with stroke and those without, and their relationship with the prognosis of stroke.Methods:A retrospective analysis was performed on medical records of 330 BP inpatients in the First Affiliated Hospital of Zhengzhou University from September 2012 to April 2020. These patients were divided into BP + stroke (ST) group and BP - ST group according to whether they were accompanied by stroke, and clinical manifestations and relevant laboratory examination results were compared between the two groups. According to the stroke outcome score assessed by modified Rankin Scale (mRS), patients in the BP + ST group were further divided into good-prognosis ST group (mRS ≤ 2 points) and poor-prognosis ST group (mRS > 2 points), and subgroup analysis was conducted. Correlations between measurement data (such as age, disease course and laboratory examination results) and mRS scores were analyzed.Results:In the BP - ST group (256 cases), 151 were males and 105 were females, and their age ranged from 19 to 92 (66.8 ± 13.6) years; in the BP + ST group (74 cases), 45 were males and 29 were females, and their age ranged from 48 to 92 (74.6 ± 9.6) years; Compared with the BP - ST group, the BP + ST group showed older age ( t = -5.57, P < 0.001), shorter disease course of BP ( Z = -3.07, P = 0.002), and higher anti-BP180 IgG antibody levels (215.0 [157.2, 283.1] U/ml vs. 155.0 [63.9, 279.8] U/ml; Z = -2.12, P = 0.034). The distribution of skin lesions significantly differed between the two groups ( χ2 = 10.51, P = 0.015), and the BP + ST group showed a significantly lower proportion of patients with generalized lesions ( P<0.05), but a higher proportion of patients with lesions on the limbs ( P<0.05). Subgroup analysis showed significant differences in the patients′ age, BP course, lesion distribution and anti-BP180 IgG antibody levels among the good-prognosis ST group, poor-prognosis ST group and BP - ST group ( F = 10.83, P<0.001; Z = 17.24, P<0.001; χ2 = 15.57, P = 0.026; Z = 6.29, P = 0.043, respectively). There was no significant difference in the age between the good-prognosis ST group and poor-prognosis ST group (adjusted P = 1.000), but the patients were significantly older in the two above groups than in the BP - ST group (adjusted P = 0.001, 0.007, respectively) ; the poor-prognosis ST group showed significantly shorter BP courses (adjusted P = 0.016, < 0.001, respectively) and a higher proportion of patients with lesions on the limbs (both P < 0.05) compared with the good-prognosis ST group and BP - ST group, and significantly higher serum anti-BP180 IgG antibody levels compared with the BP - ST group (226.2 [163.6, 285.8] U/ml vs. 155.0 [63.9, 279.8] U/ml; adjusted P = 0.037). There were no significant differences in the gender distribution, lesional morphology, percentages and counts of peripheral blood eosinophils, serum total IgE levels, and anti-BP230 IgG antibody levels between the BP + ST group and BP - ST group (all P > 0.05), or among the good-prognosis ST group, poor-prognosis ST group and BP - ST group (all P > 0.05). Correlation analysis in the BP + ST group showed a significantly negative correlation between the BP course and mRS scores ( r = -0.33, P = 0.004), and a significantly positive correlation between the anti-BP180 IgG antibody levels and mRS scores ( r = 0.34, P = 0.032) . Conclusion:There were differences in the patients′ age, BP course, lesion distribution, and anti-BP180 IgG antibody levels between the BP patients with stroke and those without, and the differences were more obvious between the poor-prognosis ST group and BP - ST group.

To report the first case of chronic disseminated paracoccidioidomycosis in China. A 49-year-old male patient presented with papules and nodules of the skin for 1 year, and papules and ulcers on the oral mucosa for 2 months. Skin examination showed the edema of the left foot, multiple crusting ulcers on the sole of the left foot, ulcers with a granular base in the interdigital regions between the third and fourth toes as well as fourth and fifth toes of the left foot, accompanied by punctate hemorrhage and exudation; there were multiple papules, nodules, and plaques on the dorsum and medial side of the left foot and the left knee, with ulcers and crusts in the center; 2 papules were observed on the left wrist, and 1 papule on the left upper lip with a crusted surface; red plaques with ulcers and punctate hemorrhage were observed on the gingival mucosa, buccal mucosa, labial mucosa, and palate, and the lesions mainly occurred on the left side. Ultrasonography of superficial lymph nodes showed bilateral cervical and supraclavicular lymph node enlargement, which was more obvious on the left side. Computed tomography of the chest and abdomen showed diffuse miliary nodular shadows, and cordlike, cloudy flocculent and nodular high-density shadows in both lungs, as well as obvious thickening of the left adrenal gland in the abdomen. Yeast cells were observed by immunofluorescent staining of biopsy tissues from the oral mucosa and left lower limb. Histopathological examination of biopsy tissues from the oral mucosa and left lower limb showed granulomatous inflammation, and refractive double-membrane yeast cells could be observed inside or outside the multinucleated giant cells, without or with a single bud or multiple buds; periodic acid-Schiff staining and hexamine silver staining of the above biopsy tissues were positive. Fungal culture of the left lower limb lesion in Sabouraud dextrose agar medium at 25℃ and 37℃ both yielded fungal hyphae. Metagenomics sequencing of the oral mucosal tissue and alveolar lavage fluid indicated the infection with Paracoccidioides brasiliensis. The diagnosis of chronic disseminated paracoccidioidomycosis was confirmed. After 1-month oral treatment with itraconazole capsules at a dose of 400 mg/d, the lesions on the skin and oral mucosa markedly improved, and computed tomography imaging of the lung and left adrenal gland also showed obvious improvement. The dose of itraconazole was reduced to 200 mg/d after 3 months. The patient′s condition further improved during a 10-month follow-up.