BACKGROUND:Croton cream can activate ERK pathways and have anti-apoptotic effects on neuronal cells.It is not clear whether it synergistically exerts anti-apoptotic effects by inhibiting the activation of JNK and p38 pathways. OBJECTIVE:To explore the effects and mechanisms of croton cream on neuronal damage and apoptosis in the ischemic cortex of rats with cerebral ischemia-reperfusion injury. METHODS:(1)Ninety Sprague-Dawley rats were randomly divided into sham operation group,model group,croton cream low-dose group,croton cream medium-dose group,croton cream high-dose group and nimodipine group,with 15 rats in each group.Except for the sham operation group,animal models of middle cerebral artery occlusion were prepared in rats by the thread method.Rats in the three croton cream groups were given 20,40,and 60 mg/kg croton cream,respectively.Rats in the sham operation and model groups were given the same amount of normal saline,once a day,for 7 consecutive days.The optimal concentration of croton cream,namely the high dose of croton cream,was selected based on neurological deficit score,TTC staining,brain tissue water content,hematoxylin-eosin staining and Nissl staining.(2)Another 120 Sprague-Dawley rats were randomly divided into sham operation group,model group,croton cream group,JNK inhibitor group,croton cream+JNK inhibitor group,p38 MAPK inhibitor group,croton cream+p38 MAPK inhibitor group,and nimodipine group,with 15 rats in each group.Animal models of middle cerebral artery occlusion were prepared using the thread method in all the groups except in the sham operation group.Thirty minutes before modeling,10 μL of SP600125(JNK inhibitor)and 10 μL of SB203580(p38 MAPK inhibitor)were injected into the lateral ventricle of the rats,respectively.Rats in croton cream groups were intragastrically given 60 mg/kg croton cream.Seven days later,the JNK/p38 MAPK signaling pathway,apoptosis-related proteins and cell apoptosis were detected by western blot,TUNEL staining and flow cytometry,respectively. RESULTS AND CONCLUSION:(1)Compared with the sham operation group,neurological deficit score,cerebral water content,cerebral infarction volume and apoptosis rate were significantly increased in the model group(P<0.05),where nerve cells showed scattered distribution.Compared with the model group,neurological deficit score,water content of brain tissue and cerebral infarction volume were significantly decreased in the croton cream medium-dose group,high-dose group and nimodipine group(P<0.05),and the pathological morphology of nerve cells was significantly improved.(2)Compared with the JNK inhibitor group,p-JNK/JNK,p-p38/p38 and Bax expressions in rat brain tissue and the apoptotic rate were significantly decreased in the croton cream+inhibitor groups(P<0.05),while the expression of and Bcl-2 was significantly increased(P<0.05).To conclude,croton cream may inhibit the activation of JNK/p38 MAPK signaling pathway and reduce neuronal apoptosis to achieve neuroprotective effects in rats with cerebral ischemia-reperfusion injury.

Objective:To explore and analyze the influence of thoracic types and symmetry methods on the treatment effect before operation for the breast cancer patients undergoing immediate breast reconstruction.Methods:From January to October 2018, 30 female patients with unilateral breast cancer, aged 33-48 (39.6±4.6) years, were treated in the Breast Center of Qingdao University Affiliated Hospital. Thoracic CT was used to analyze thoracic types and symmetry. The measurement indexes included the maximum transverse diameter of thorax, the maximum longitudinal diameter of left and right sides, the ratio of the maximum transverse longitudinal diameter of left and right sides, the anterior chest angle of left and right sides, the posterior chest angle and the external chest angle, and the type and symmetry of thorax were analyzed, and the choice of prosthesis was adjusted according to the measurement results.Results:All the 30 patients recovered smoothly, and no complications such as infection and prosthesis removal occurred. Anatomical prostheses were selected in 22 patients, including 12 cases with medium high protrusion prosthesis, 10 cases with low protrusion prosthesis and 8 cases with round low protrusion prosthesis. The average follow-up time was 15 months after operation. The postoperative follow-up photos were evaluated aesthetically by using the BRAST-Q questionnaire and 3 non-operative doctors, among which 22 cases were excellent, 4 cases were good, 3 cases passed and 1 case was poor. Excellent and good rate was 86.7%. Among them, 3 cases were satisfactory, 1 case was scar hyperplasia of local incision, 2 cases were mild capsular contracture, and 1 case was subject to prosthesis displacement.Conclusions:The measurement and analysis of thoracic type and symmetry before operation can help to adjust the choice of prosthesis, check the obvious thoracic deformity and avoid affecting the surgical results.

Objective: To investigate the effect of local autologous dermal fibroblasts transplantation on hypertrophic scar formation and wound healing quality in early scar formation. To explore the feasibility of fibroblasts for prevention and treatment of hypertrophic scar. Methods: Dermal fibroblasts were isolated from the dorsal skin tissue of New Zealand white rabbits by mechanical method combined with enzyme digestion. Passage 3 cells were induced to differentiate into osteoblasts and adipocytes. The complete epithelialization time and hypertrophic scar formation after full-thickness skin defect were confirmed by pre-experiment study. In the experiment, 6 rabbits were used, left ear as experimental group and right ear as control group. In the experimental group, the passage 4 dermal fibroblasts labeled with 5-bromodeoxyuridine (5-BrdU) were injected subcutaneously around the wound and hypertrophic scar on 20 d (day 2 after epithelialization) and 30 d (most obvious scar hyperplasia) after surgery. As a control group, physiological saline was injected following the same protocol. On 37 days after surgery, the hypertrophic scar tissue were harvested and assessed by gross view and histological examination. The transplanted cells were detected by immunofluorescence staining, transforming growth factor beta 1 (TGF-β1) and decorin(DCN) mRNA expression were assayed by real-time fluorescence polymerase chain reaction (RT-PCR), and the protein expression of TGF-β1、DCN、Collagen type Ⅰ and type Ⅲ were tested by enzyme linked immunosorbent assay(ELISA). Results: Compared with the control group, the scar in the experimental group was flatter and softer, the color was slightly lighter, and the volume was reduced. The histological results showed that compared with the control group, the number of fibroblasts and inflammatory cells in the superficial dermis was reduced, the proliferation of connective tissue and collagen deposition were reduced, and the basal cells and collagen fibers were arranged in order in the experimental group. The results of RT-PCR showed that TGF-β1 mRNA expression level in the hypertrophic scar tissue reduced significantly and DCN increased significantly in the experimental group, compared with the control group (P<0.01). ELISA results showed that TGF-β1, type Ⅰ and Ⅲ collagen contents in hypertrophic scar tissue were significantly lower than that in the control group, while the DCN expression was significantly increased (P<0.01). Conclusions The rabbit dorsal dermal fibroblasts significantly inhibited the hypertrophic scar in rabbit ears. The mechanism may be related to the up-regulation of DCN expression and down-regulation of TGF-β1 expression, reduction of type Ⅰ and Ⅲ collagen synthesis and extracellular matrix deposition, in the hypertrophic scar tissue.

Objective To conduct the comparative study of the measurement of breast volume by MRI imaging and Archimedes method in immediate implant breast reconstruction.Methods A total of 44 patients who were diagnosed as breast cancer and undergone immediate implant breast reconstruction were selected from March 2011 to March 2013.22 cases were guided to select suitable breast implants by measuring the breast volume and correlative radial line based on MRI imaging.Control group containing 22 cases were guided to select breast implants by traditional Archimedes method and clinical experience.3 breast surgeons and plastic surgeons who did not participate in the operation were selected to judge the postoperative breast shape.Data of two groups were analyzed by using chisquare test.Results All the patients recovered smoothly after operation without infection,and the wound healed well.The breast shape was evaluated.21 cases (95.5 ％) were good in test group and 1 case (4.5％) was poor;16 cases (72.7％) were good in control group and 6 cases (27.3) were poor.The comparison between the test group and control group had statistical significance (P＜0.05).Conclusions The method to measure the breast volume and correlative radial line based on MRI imaging has important values for selecting breast implants in immediate implant breast reconstruction.It could be extensively used in clinical practice.

Objective To survey the dietary iodine intake among residents in the coastal province of Fujian after universal salt iodization. Methods Using multi-stage stratified random sampling method, four different geographic areas of coastal cities, inland cities, coastal rural and inland rural areas were selected; 3 counties (cities, districts) were selected from each area; 30 families were selected from each of those counties (cities, districts). Total dietary study method was applied to investigate the basic information, all food and water intake during the investigation, the spice variety, quantity and cooking methods, and recipes. All foods consumed were divided into 13 classes:grains and their products, beans(including nuts) and their products, potato and its products, meat and its products, eggs and their products, aquatic products, milk and dairy products, vegetables, fruits and their products, sugar and sugar products, beverage and water, alcohol and seaweeds. Food samples were collected and iodine contents of different types of food were measured, and daily food consumption of the reference man(adult male) in the four areas was calculated. Then the male adult dietary iodine intake in the four areas and Fujian Province was estimated. Results A total of 69 families(accounting for 19.2% of the 359 families surveyed) ate seaweed foods during the three days survey period. The average salt intakes per reference man were 4.5, 5.8, 7.5, 12.3 g, respectively, in coastal cities, inland cities, coastal rural and inland rural. The adult male dietary iodine intake in Fujian Province was 343.8μg/d, and the values were 234.4, 347.1, 328.4, 465.3μg/d, respectively, in coastal cities, coastal rural, inland cities and inland rural. If people did not eat seaweed foods, the results would be 239.7, 137.5, 253.6, 200.3, 367.4μg/d. The main sources of dietary iodine intake were vegetables (33.2%), seaweeds(30.3%), meat(13.4%) and aquatic products(13.0%). If people did not eat seaweed foods, the results would be vegetables(47.6%), meat(19.1%) and aquatic products(18.7%), and iodine contribution rate of the 3 types of food was 85.4%. Conclusions Under the conditions of eating iodized salt, male adult dietary iodine intake is appropriate in different areas of Fujian Province. There is no such problem as iodine excessive. Iodized salt is the main source of dietary iodine. Residents in coastal city, if not eating seaweed foods, will be at risk of iodine deficiency, even if they eat iodized salt.

Objective To probe the teaching method of application of problem-based learning (PBL)with computer aided technology in teaching of breast surgery clinical practice.Methods 48 students were randomly divided into two groups:control group(n=24) were taught by lecture-based learning(LBL),experimental group (n=24) were taught by method of application of problem-based learning(PBL)with computer aided technology.Questionnaire was provided to the students of experimental group to investigate the degree of recognition to the teaching method of problem-based learning (PBL)with computer aided technology.5 patients from experimental group and 5 patients from control group participating in teaching were given questionnaires to investigate if their right of privacy were violated.The theory examination score(out of 50) and comprehensive skills assessment score (out of 50) were compared between the two groups; the mean differences between the two groups were compared by t test.Result The teaching method of application of problem-based learning(PBL) with computer aided technology was recognition by 91.7 ％(22 persons) of students.20 percent(one person) of the experimental group patients felt their right of privacy violated; 60 percent(three persons) of the control group patients felt their right of privacy violated.The theory test scores of the experimental group and control group were (46.23 ± 2.45) and (45.52 ± 3.29).There was no statistical significance (t=1.230,P=0.225).The skill test scores of the experimental group and control group were(47.23± 2.45) and (36.52 t 3.29).There was statistical significance(t=12.791,P=0.000).Conclusion The teaching method of application of problem-based learning(PBL) with computer aided technology can help encourage the study interest of students,and cultivate their clinical thinking method and capacity,and protect the right of privacy of the patients,avoiding patient-hospital disputes.

Objective To explore the causes of secondary breast deformity after breast conservative therapy and its preventive methods.Methods 30 cases were selected from the patients with secondary breast deformity of more than 1 year after breast conservative therapy,and more than half a year after radiotherapy and chemotherapy,and no local recurrence and distant metastasis occurred.After cicatrectomy and contracture loose solution,different methods were used according to varying degrees of the secondary breast deformity to reconstruct and repair the deformity.Results Three reconstructive methods were conducted.Local mammary flaps were used in 7 patients,latissimus dorsi myocutaneous flap in 21 patients,and transverse rectus abdominal musculocutaneous flaps in 2 patients.The shape and feeling were satisfactory in 30 patients with reconstructive breasts,in which 23were excellent (76.7 ％),and 7 were good (23.3 ％).All the patients were followed-up for average 2.5 years,and they were survived without tumor recurrence.Conclusions The secondary breast deformity after breast conservative therapy could be effectively prevented,and successfully reconstructed and repaired with different procedure according to varying degrees of the secondary breast deformity.As a result,the shape and feeling of reconstructive breasts are satisfactory.

Six recombinant plasmids co-expressing the wild-type GP5 gene or the codon-optimized GP5 gene (containing pan-DR epitope) of porcine reproductive and respiratory syndrome virus (PRRSV) and the E2 gene of classical swine fever virus (CSFV) or the E2 fused with the UL49 of pseudorabies virus (PrV) were constructed based on the suicidal DNA vaccine pSFV1CS-E2 described previously. Expression of GP5 and E2 was confirmed by indirect immunofluorescence assay. The immunogenicity of six plasmids was evaluated in BALB/c mouse model. For the six plasmids, low-level of E2 and GP5 protein specific antibodies could be detected in the sera of the immunized mice. Specific lymphoproliferative responses to the PRRSV or CSFV stimulation were induced in the splenocytes of the immunized mice as demonstrated by CFSE staining assay and WST-8 assay. Antigen specific IFN-gamma and L-4 secretion was detected in the splenocytes of some immunized mice by cytokine ELSIA. Fusion with the PrV UL49 in the suicidal vaccines induced significantly higher lymphoproliferative responses and cytokine secretion. Taken together, the suicidal DNA vaccines co-expressing GP5 and E2 could induce PRRSV and CSFV specific humoral and cell-mediated immune responses.
Animals ; Antibodies, Viral ; blood ; Antibody Formation ; Cytokines ; blood ; Female ; Immunity, Cellular ; Lymphocytes ; immunology ; Mice ; Mice, Inbred BALB C ; Random Allocation ; Vaccines, DNA ; biosynthesis ; immunology ; Viral Envelope Proteins ; genetics ; immunology ; Viral Structural Proteins ; genetics ; immunology ; Viral Vaccines ; biosynthesis ; immunology

AIM: Tranditional methods of screening drugs for benign prostatic hyperplasia(BPH)requires senile male animals such as dogs or rats.It consumes a long time to get the results. Over-expression of type Ⅱ5α-reductase in prastate induces BPH.A fast and efficient screening model of type Ⅱ5α-reductase inhibitors for BPH was set up in this paper.METHODS:Microsomes were extracted from male Sprague-Dawley rat livers by gradient centrifugation.Type Ⅱ5α-reductase enzyme-catalyzed reaction was assayed by UV-spectrophotometry using testosterone as a substrate and NADPH as hydrogen donor.The change of enzymatic activity was recorded with a NADPH wavelength of 340 nm by subtracted descending velocity of the control(without 5α-reductase).Effects at different conditions(temperatures,pH,enzyme and testosterone concentrations)on 5α-reductase were assayed.RESULTS:The suitable condition of type Ⅱ5α-reductase reaction Was defined as concentration of 109.05 mg protein/L enzyme(pH 6.00)with 2 μmol/L testosterone at 37℃.Michaelis'constant of type Ⅱ5α-reductase was 0.6μmol/L.Finasteride,a new drug for BPH,significantly inhibited activity of type Ⅱ 5α-reductase.IC50 of finasteride Was 64.1 nmol/L.As solvent of drugs,concentration of ethanol below 1.1% did not inhibite enzymatic activity(P＞0.05).Concentration of ethanol above 1.6%could obviouslv suppress enzymatic activity(P＜0.01).Daytime difference within five days had no significant difference(P＞0.05).CONCLUSION:A handy and fast screening method for type Ⅱ 5α-reductase inhibitors has been set up using UV-spectrophotometry.It may be used to screening drugs for BPH treatment.

BACKGROUND: Extended high frequency audiometry is able to detect early cochlea damage. So it is useful in monitoring ototoxic, senile and noise induced hearing loss.OBJECTIVE: To study with extended high frequency audiometry the auditory function of people in highland of qinghai province without noise pollution and evaluate the sensitivity of this method in diagnosing early noise induced hearing loss. To provide the epidemiological data for early control of noise induced hearing loss in this area.DESIGN:It is a transverse comparison among normal people in two areas.SETTING: This study was cooperated by the Otolaryngeal Department of Gulou Hospital affiliated to Nanjing Medical College, the Otolaryngeal Department of Xijing Hospital of the Fourth Medical College and Auditory Research Center of Giessen University, Germany.PARTICIPANTS: Totally 300 people lived in highland of qinghai province received hearing test including extended high frequency audiometry. At the same time another 300 people were chosen in Nanjing city as control group.METHODS: The subjects in the two groups received the whole set of hearing tests including extended high frequency audiometry and answered a questionnaire about their personal history. The mean hearing thresholds from 0. 25-16 kHz between the two groups were compared with t-test.MAIN OUTCOME MEASURES: The difference of mean hearing thresholds from 0. 25-16 kHz between the two groups.RESULTS: There was no significant difference between hearing threshold of low frequencies and speech frequencies. The hearing threshold of high frequencies rose with the stepping-up of testing frequencies which made the possibility of measuring the hearing threshold reduced. But the hearing threshold above 6 kHz of the people in area without noise pollution was significantly higher than that of the control( P ＜ 0.05) . No difference between children and adolescence was found in the 2 groups. The epidemiological data revealed that the people without noise pollution had unperceptible hearing damage compared with the control group.CONCLUSION: The damage of impulsive noise to hearing began at 20 kHz and developed from 20 kHz to 10 kHz. If it was not protected, it would be extended to the speech frequencies and the hearing loss would get worse. So the extended high frequency audiometry was thought helpful to screen noise induced hearing loss early.