Plantar Pressure and Offloading Insole Structure for Diabetic Patients: A Finite Element Analysis / 中国康复理论与实践

Objective:To investigate the plantar pressure features of diabetic patients, and design the offloading structure of insole to reduce the plantar pressure and internal stress of the soft tissue. Methods:A three-dimensional finite element model of foot was established based on CT images. Hole structure was designed in the high plantar pressure area of diabetic patients. The effects of diameter, depth and interval of holes on plantar pressure was analyzed through orthogonal test and finite element analysis, to obtain the optimal scheme; and the offloading effect was analyzed with finite element analysis and experiment. Results:The peak plantar pressure was higher in diabetic patients than in healthy individuals. The holes with 5 mm diameter, 6 mm depth and 2 mm interval in metatarsal and calcaneus regions might effectively reduce the plantar pressure and internal stress of soft tissue, which was 15.6%, 45.6%, 53.5% and 10.1% less of the peak plantar pressure on toes, metatarsal, midfoot and calcaneus area, respectively, compared to walking without insoles. Conclusion:Finite element analysis is helpful to explore the internal stress of soft tissue in diabetic patients, and insole with hole structure can reduce the plantar pressure and internal stress of soft tissue.

Finite element analysis of the effects of periodontal tissue temperature by continuous wave technique / 华西口腔医学杂志

OBJECTIVES@#The safety of root canal filling with 200 °C hot gutta-percha was investigated to study the effect of continuous wave technique combined with high-temperature injectable gutta-percha condensation technique on the surface temperature of periodontal tissue.@*METHODS@#CT technique and Mimics, Geomagic, and Solidworks software were utilized to build the entity models of alveolar bone, dentin and root canal, periodontal ligament, and blood flow, respectively, which were then assembled in Solidworks into a finite element model of tooth with blood flow. By utilizing ABAQUS collaborative simulation platform, fluid-structure coupling was analyzed on the whole process of root canal filling. Consequently, the surface temperature of the periodontal tissue was obtained.@*RESULTS@#In the absence of blood flow, the temperature of the periodontal ligament surface reached 50.048 ℃ during root canal filling with 200 ℃ gutta-percha. Considering blood flow, the temperature of periodontal ligament surface was 39.570 ℃.@*CONCLUSIONS@#The temperature of the periodontal ligament surface increased when the continuous wave root canal was filled with 200 ℃ gutta-percha, and the periodontal tissue was not damaged.

Hippocampus chronic deep brain stimulation induces reversible transcript changes in a macaque model of mesial temporal lobe epilepsy / 中华医学杂志(英文版)

BACKGROUND@#Deep brain stimulation (DBS) has seizure-suppressing effects but the molecular mechanisms underlying its therapeutic action remain unclear. This study aimed to systematically elucidate the mechanisms underlying DBS-induced seizure suppression at a molecular level.@*METHODS@#We established a macaque model of mesial temporal lobe epilepsy (mTLE), and continuous high-frequency hippocampus DBS (hip-DBS) was applied for 3 months. The effects of hip-DBS on hippocampus gene expression were examined using high-throughput microarray analysis followed by bioinformatics analysis. Moreover, the microarray results were validated using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analyses.@*RESULTS@#The results showed that chronic hip-DBS modulated the hippocampal gene expression. We identified 4119 differentially expressed genes and assigned these genes to 16 model profiles. Series test of cluster analysis showed that profiles 5, 3, and 2 were the predominant expression profiles. Moreover, profile 5 was mainly involved in focal adhesion and extracellular matrix-receptor interaction pathway. Nine dysregulated genes (Arhgap5, Col1a2, Itgb1, Pik3r1, Lama4, Fn1, Col3a1, Itga9, and Shc4) and three genes (Col1a2, Itgb1, and Flna) in these two pathways were further validated by qRT-PCR and Western blot analyses, respectively, which showed a concordance.@*CONCLUSION@#Our findings suggest that hip-DBS could markedly reverse mTLE-induced abnormal gene expression. Findings from this study establish the basis for further investigation of the underlying regulatory mechanisms of DBS for mTLE.

Clinical information and management status of de novo stage IV breast cancer patients: a Chinese multicenter investigation (CSBrS-002) / 中华医学杂志(英文版)

BACKGROUND@#Although de novo stage IV breast cancer is so far incurable, it has entered an era of individualized treatment and chronic disease management. Based on systemic treatment, whether the surgical resection of primary or metastatic foci of de novo stage IV breast cancer can bring survival benefits is currently controversial. We aimed to explore the clinicopathological factors and current status of the management of de novo stage IV breast cancer in China to provide a reference for clinical decisions.@*METHODS@#Based on the assistance of Chinese Society of Breast Surgery, a retrospective study was conducted to analyze the clinical data of patients with de novo stage IV breast cancer in 33 centers from January 2017 to December 2018. The relationship between basic characteristic (age, menstrual status, family history, reproductive history, pathological type, estrogen receptor [ER] status, progesterone receptor [PR] status, human epidermal growth factor receptor 2 [HER2] status, Ki-67 percentage, and molecular subtype), and metastasis sites (lung metastasis, liver metastasis, and bone metastasis) was examined by Pearson Chi-square tests.@*RESULTS@#A total of 468 patients with de novo stage IV breast cancer were enrolled. The median age of the enrolled patients was 51.5 years. The most common pathological type of primary lesion was invasive carcinoma (97.1%). Luminal A, luminal B, HER2 overexpressing, and triple-negative subtypes accounted for 14.3%, 51.8%, 22.1%, and 11.8% of all cases, respectively. Age, PR status, and HER2 status were correlated with lung metastasis (χ2 = 6.576, 4.117, and 8.643 and P = 0.037, 0.043, and 0.003, respectively). Pathological type, ER status, PR status, and molecular subtype were correlated with bone metastasis (χ2 = 5.117, 37.511, 5.224, and 11.603 and P = 0.024, <0.001, 0.022, and 0.009, respectively). Age, PR status, HER2 status, Ki-67 percentage, and molecular subtype were correlated with liver metastasis (χ2 = 11.153, 13.378, 10.692, 21.206, and 17.684 and P = 0.004, <0.001, 0.001, <0.001, and 0.001, respectively). Combined treatment with paclitaxel and anthracycline was the most common first-line chemotherapy regimen for patients with de novo stage IV breast cancer (51.7%). Overall, 91.5% of patients used paclitaxel-containing regimens. Moreover, 59.3% of hormone receptor-positive patients underwent endocrine therapy.@*CONCLUSIONS@#In 2018, 1.07% of patients from all studied centers were diagnosed with de novo stage IV breast cancer. This study indicated that 95.1% of patients received systemic therapy and 54.2% of patients underwent surgical removal of the primary lesion in China.

Clinical Characteristics and Genetic Analysis of Klippel-Feil Syndrome / 中国医学科学院学报

Objective To summarize clinical characteristics and investigate possible pathogenic gene of Klippel-Feil syndrome(KFS)by the self-designed multigene panel sequencing,so as to decipher the molecular basis for early diagnosis and targeted therapy.Methods From January 2015 to December 2018,we consecutively recruited 25 patients who were diagnosed with KFS in Peking Union Medical College Hospital.The demographic information,clinical manifestations,physical examination and radiological assessments were analyzed.Multigene panel sequencing was performed after DNA extraction from peripheral blood.The possible pathogenic mutations of KFS were explored on the basis of bioinformatics analysis.Results The KFS cohort consisted of 25 patients,including 15 males and 10 females,with a mean age of(12.9±7.3)years.Limited cervical range of motion was the most common clinical feature(12 cases,48%).Based on the Samartzis classification,the proportion of patients suffered from short neck(P=0.031)and limited cervical range of motion(P=0.026)in type Ⅲ KFS was significantly higher than that in type Ⅱ and type Ⅰ KFS.Panel sequencing detected a total of 11 pathogenic missense mutations in eight patients,including COL6A1,COL6A2,CDAN1,GLI3,FLNB,CHRNG,MYH3,POR,and TNXB.There was no pathogenic mutation found in five reported pathogenic genes(GDF6,MEOX1,GDF3,MYO18B and RIPPLY2)associated with KFS.Conclusions Our study has shown that patients with multiple contiguous cervical fusions are more likely to manifest short neck,limited cervical range of motion,and clinical triad.Therefore,these patients need additional attention and follow-up.Our analysis highlights novel KFS-related genetic variants,such as COL6A and CDAN1,extending the spectrum of known mutations contributing to this syndrome and providing a basis for elucidating the pathogenesis of KFS.

Anterior thalamic nuclei deep brain stimulation inhibits mossy fiber sprouting via 3',5'-cyclic adenosine monophosphate/protein kinase A signaling pathway in a chronic epileptic monkey model / 中华医学杂志(英文版)

BACKGROUND@#Anterior thalamic nuclei (ATN) deep brain stimulation (DBS) is an effective method of controlling epilepsy, especially temporal lobe epilepsy. Mossy fiber sprouting (MFS) plays an indispensable role in the pathogenesis and progression of epilepsy, but the effect of ATN-DBS on MFS in the chronic stage of epilepsy and the potential underlying mechanisms are unknown. This study aimed to investigate the effect of ATN-DBS on MFS, as well as potential signaling pathways by a kainic acid (KA)-induced epileptic model.@*METHODS@#Twenty-four rhesus monkeys were randomly assigned to control, epilepsy (EP), EP-sham-DBS, and EP-DBS groups. KA was injected to establish the chronic epileptic model. The left ATN was implanted with a DBS lead and stimulated for 8 weeks. Enzyme-linked immunosorbent assay, Western blotting, and immunofluorescence staining were used to evaluate MFS and levels of potential molecular mediators in the hippocampus. One-way analysis of variance, followed by the Tukey post hoc correction, was used to analyze the statistical significance of differences among multiple groups.@*RESULTS@#ATN-DBS is found to significantly reduce seizure frequency in the chronic stage of epilepsy. The number of ectopic granule cells was reduced in monkeys that received ATN stimulation (P < 0.0001). Levels of 3',5'-cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) in the hippocampus, together with Akt phosphorylation, were noticeably reduced in monkeys that received ATN stimulation (P = 0.0030 and P = 0.0001, respectively). ATN-DBS also significantly reduced MFS scores in the hippocampal dentate gyrus and CA3 sub-regions (all P < 0.0001).@*CONCLUSION@#ATN-DBS is shown to down-regulate the cAMP/PKA signaling pathway and Akt phosphorylation and to reduce the number of ectopic granule cells, which may be associated with the reduced MFS in chronic epilepsy. The study provides further insights into the mechanism by which ATN-DBS reduces epileptic seizures.

125Ⅰ seed brachytherapy for recurrent salivary gland carcinoma after external radiotherapy / 北京大学学报(医学版)

OBJECTIVE@#To investigate the clinical application and efficacy of 125Ⅰ radioactive seeds implantation in the treatment of recurrent salivary gland carcinoma after external radiotherapy.@*METHODS@#From July 2004 to July 2016, 43 cases of recurrent salivary gland carcinoma of the neck after external radiotherapy or surgery combined with external radiotherapy were treated. According to the conventional segmentation radiotherapy for head and neck cancer (once a day, 1.8-2.0 Gy each time, 5 days per week), the cumulative radiation dose of the patients in this group was calculated. In the study, 26 patients received 50-60 Gy, 7 patients received less than 50 Gy, 4 patients received 60-70 Gy, and 6 patients received more than 80 Gy (range: 80-120 Gy). The interval between the last external irradiation and local recurrence was 4-204 months, and the median interval was 48 months. Among them, 25 cases were treated with 125Ⅰ radioactive seeds implantation only and 18 cases were treated with 125Ⅰ radioactive seeds implantation after operation. The prescription dose was 100-140 Gy. The control rate, survival rate and disease-free survival rate were recorded to evaluate the side effects.@*RESULTS@#The median follow-up time was 27 months (ranging from 2.5 to 149.0 months). Among them, the median follow-up time of adenoid cystic carcinoma patients was 31 months (range: 2.5-112.0 months), and the median follow-up time of mucoepidermoid carcinoma patients was 18 months (range: 5-149 months). The local control rates for 1, 3 and 5 years were 66.5%, 48.8% and 42.7%, respectively. The 1-, 3- and 5- year survival rates were 88.0%, 56.7% and 45.8%, respectively. The disease-free survival rates of 1, 3 and 5 years were 58.3%, 45.4% and 38.1%, respectively. There was no statistically significant difference in local control rate, survival rate, and disease-free survival between the radioactive seeds implantation group and the radioactive seeds implantation group after surgical resection. There were 2 cases of acute radiation reaction Ⅰ/Ⅱ and 3 cases of reaction Ⅲ or above. In the late stage of radiotherapy, there were 8 cases with Ⅰ/Ⅱ grade reaction and 3 cases with Ⅲ grade or above reaction. The incidence of radiation reactions of Grade Ⅲ and above was 7%.@*CONCLUSION@#125Ⅰ radioactive seeds implantation provides an alternative method for the treatment of recurrent salivary gland carcinoma after external radiotherapy. The local control rate and survival rate are improved on the premise of low incidence of side effects.

wareness and knowledge of oral cancer among 1 483 residents in Beijing / 北京大学学报(医学版)

OBJECTIVE@#To investigate the oral cancer awareness, its related knowledge among residents in Beijing.@*METHODS@#A questionnaire survey was conducted among Beijing residents (non-medical related personnel) on the knowledge of oral cancer and its prevention and treatment.@*RESULTS@#A total of 1 483 questionnaires were completed, including 663 males (44.6%) and 820 females (55.3%), ranging from 15 to 91 years. Lung cancer was the most mentioned as heard of (94.3%), followed by liver cancer (92.5%) and gastric cancer (92.4%), while oral cancer was the least heard of (47.7%). Family account (P=0.015), residence (P=0.028), income (P=0.024), frequency of brushing teeth (P<0.001), frequency of mouth self-examination (P<0.001) and chew betel nut chewing (P=0.015) were significantly associated with oral cancer awareness. Most respondents learned about oral cancer from TV programs (24.3%), phone news (22.0%), WeChat (11.3%) and chatting with friends or people around (14.6%). Only 34.9% knew that smoking was a risk factor of oral cancer, and this knowledge was associated with age (P=0.011), education level (P=0.007), frequency of brushing teeth (P<0.001), and frequency of mouth self-examination (P=0.002). Only 23.5% knew that drinking was a risk factor of oral cancer, and this knowledge was associated with education level (P=0.002), residence (P=0.022), frequency of brushing teeth (P=0.009), and frequency of mouth self-examination (P=0.005). Only 35.1% knew that betel nut chewing was a risk factor for oral cancer, and this knowledge was associated with age (P=0.040), education level (P=0.002), family account (P=0.002), income (P=0.005), frequency of brushing teeth (P=0.001), frequency of mouth self-examination (P<0.001), and betel nut chewing (P=0.002). Only 35.7%, 21.6% and 16.9%, respectively, knew that long-term unhealed ulcers, white plaques and red plaques in the mouth were the early signs of oral cancer.@*CONCLUSION@#This survey demonstrates a general lack of public awareness and knowledge about signs and symptoms of oral cancer, and its risk factors and treatment. Some residents have not formed good oral hygiene habits. Specific measures should be taken to improve the public awareness of oral cancer and related knowledge about prevention and treatment, such as the use of various new media for the public to promote their oral health knowledge, in order to reduce the incidence of oral cancer, improve early attendance rates and the patients' survival rates.

Treatment of early-onset scoliosis: techniques, indications, and complications / 中华医学杂志(英文版)

The treatments for early-onset scoliosis (EOS) remain great challenges for spine surgeons. This study aimed to comprehensively review the treatments for EOS, especially the advancements made in the last decade. Current studies on EOS were retrieved through a search on PubMed, UpToDate, the Web of Science Core Collection and Scopus were reviewed. The most pertinent information related to the current treatments for EOS was collected. The foci of treatments for EOS have included creating a well-developed thoracic cavity, improving lung volume, and improving pulmonary function. Conservative treatments include bracing, casting, halo-gravity traction, and physiotherapy. Serial casting is the most effective conservative treatment for EOS. Surgical treatments mainly include growth-friendly techniques, which are generally classified into three types according to the amount of correction force applied: distraction-based, compression-based, and growth-guided. The distraction-based systems include traditional or conventional growing rods, magnetically controlled growing rods, and vertical expandable prosthesis titanium ribs. The compression-based systems include vertebral body stapling and tethering. The growth-guided systems include the Shilla system and modern Luque trolley. In addition, some newer techniques have emerged in recent years, such as posterior dynamic deformity correction (ApiFix). For EOS patients presenting with sharp deformities in a long, congenital spinal deformity, a hybrid technique, one-stage posterior osteotomy with short segmental fusion and dual growing rods, may be a good choice. Hemivertebra resection is the gold standard for congenital scoliosis caused by single hemivertebra. Although the patient's growth potential is preserved in growth-friendly surgeries, a high complication rate should be expected, as well as a prolonged treatment duration and additional costs. Knowledge about EOS and its treatment options is rapidly expanding. Conservative treatments have specific limitations. For curves requiring a surgical intervention, surgical techniques may vary depending on the patients' characteristics, the surgeon's experience, and the actual state of the country.

Effect of siRNA-Interfering β-Catenin Expression on MDR of Human Multiple Myeloma Cell Line / 中国实验血液学杂志

OBJECTIVE@#To investigate the effecr of siRNA-interfering β-catenin expression on drug-resistance of multiple myeloma cells.@*METHODS@#The multiple myeloma cell line RPMI-8226 was cultured in vitro. The maphalan-resistant cell model was established by concentration gradient ascending of durg, then the drug-resistant cell line was instantaneously transfected with β-catenin siRNA, the sensitivity of RPMI 8226 cells to maphalan was detected by CCK-8 meltod before and after the transfection with siRNA; the mRNA and protein expression of β-catenin was detected by qRT-PCR and Western blot respectively, the apoptosis of cells was detected by flow cytometry.@*RESULTS@#IC of maphalan decreased from (5.29±0.19) μmol/L to (1.88±0.64) μmol/L, suggesting that the deplation of β-eatenin restored the sensitivity of drug-resistant cell line RPMI-8226 to malphalan. The Western blot showed that after the instaintaneous transfection with β-catenin siRNA, the β-catenin protein expression level obviously decreased, compared with level before transfection. After transfection, the maplalan-inducing apoptosis rate of cells increased from (35±0.5)% to (54±0.4)%, suggesting that the β-catinin gene may correlated with drug-resistance of cells. Interfering the expression of β-catenin gene could enhance the sensitivity of drug-resistant RPMI-8226 cells to maphalan.@*CONCLUSION@#The β-catenin siRNA interfereuce can inhisit the β-catenin gene expression in Wnt/β-catenin signaling pathway, suppress the cell proliferation, enhence the toxicity of maphalan on drug-resistant RPMI-8226 cells, thus result in increase of cell apoptosis.

Effect of Bushen Yanggu Decoction on Multidrug Resistance of Multiple Myeloma Cell Line KM3/BTZ / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of Bushen Yanggu Decoction (BYD) on drug resistance and proliferation of human multiple myeloma-resistant KM3/BTZ cells.@*METHODS@#Human multidrug-resistant KM3/BTZ cells were established by Bortezomib (BTZ) gradient induction. The effects of commonly used chemotherapeutic drugs and serum containing Bushen Yanggu Decoction (BYD) on the proliferation of KM3 cells and KM3/BTZ cells were detected by MTT assay. RT-qPCR and Western blot were used to detect the expression of Par-4, HSP27 and P-gp genes. Flow cytometry was used to detect cell apoptosis.@*RESULTS@#The established KM3/BTZ cells could produce varying degree of resistance to commonly used chemotherapeutic drugs. Among them, the highest resistance index (RI) to BTZ was 20.269. MTT assay showed that the proliferation of KM3/BTZ cells treated with serum containing Bushen Yanggu Decoction was inhibited, and the inhibitory effect increased with the serum concentration incranse of Bushen Yanggu Decoction. The serum containing Bushen Yanggu Decoction could inhibit the proliferation of KM3/BTZ cells, and induce apoptosis, significantly reduce the drug-resistance of KM3/BTZ cells, up-regulate the expression of Par-4, down-regulate the expression of HSP27 and P-gp.@*CONCLUSION@#Bushen Yanggu Decoction can effectively inhibit the proliferation of KM3/BTZ cells and induce apoptosis. Bushen Yanggu Decoction can effectively reverse the multidrug-resistance of KM3/BTZ cells. The mechanism may be related with the decrease of expression of HSP27 and P-gp and the increase of expression of Par-4.

Study of surgery combined with 125I brachytherapy for adenoid cystic carcinoma of oral and maxillofacial region / 北京大学学报(医学版)

OBJECTIVE@#To retrospectively analyze the results of treatment outcome by surgery combined with 125I brachytherapy and correlative factors of adenoid cystic carcinoma (ACC).@*METHODS@#In the study, 75 patients with primary ACC of oral and maxillofacial region were treated by surgery combined with 125I seeds brachytherapy. Radical resection or subtotal resection was applied for the tumor. The brachytherapy treatment planning system was used to create implant plans with the prescribed dose of 60 Gy to 120 Gy. The 125I seeds were implanted intraoperatively or postoperatively. The regular follow-up was required. The Kaplan-Meier method was used to assess the tumor control rate and the patients' survival rates. Meanwhile, the Cox regression analysis was used to find out the prognostic factors.@*RESULTS@#Local control rates at the end of 3 and 5 years were as follows: T1-T2, 92.2% and 82.0%; T3-T4, 82.6% and 82.6%; and overall, 90.0% and 78.8%. The disease-free survival rates were 74.9% and 54.3%, respectively. The overall survival rates for all the patients were 86.0% and 79.6%, respectively at the end of 3 and 5 years and were 91.3% and 91.3% for T1-T2 patients vs. 73.9% and 59.7% for T3-T4 patients. Distant metastasis-free survival rates at the end of 3 and 5 years were 84.4% and 76.7%, respectively. The distant metastasis-free survival rates at the end of 3 and 5 years were 83.4% and 79.6% with T1-T2 lesion compared with 86.0% and 67.8% with T3-T4 lesion. According to the COX univariate analysis and multivariate analysis, the risk of local recurrence would be raised by the age. Tumor stage and tumor site were the prognostic factors of the overall survival rates.@*CONCLUSION@#125I brachytherapy conducted as an adjuvant therapy postoperatively of ACC of oral and maxillofacial region can acquire satisfactory localregional control, distant metastasis-free survival, disease-free survival and overall survival. Tumors are prone to recur on the older patients. Patients having advanced tumor stage or tumor located in the nasal cavity or sinuses will suffer lower survival rates.

Simulation of dose distribution in bone medium of 125I photon emitting source with Monte Carlo method / 北京大学学报(医学版)

OBJECTIVE@#To present a theoretical analysis of how the presence of bone in interstitial brachytherapy affects dose rate distributions with MCNP4C Monte Carlo code and to prepare for the next clinical study on the dose distribution of interstitial brachytherapy in head and neck neoplasm.@*METHODS@#Type 6711,125I brachytherapy source was simulated with MCNP4C Monte Carlo code whose cross section library was DLC-200. The dose distribution along the transverse axis in water and dose constant were compared with the American Association of Physicists in Medicine (AAPM) TG43UI update dosimetry formalism and current literature. The validated computer code was then applied to simple homogeneous bone tissue model to determine the affected different bone tissue had on dose distribution from 125I interstitial implant.@*RESULTS@#125I brachytherapy source simulated with MCNP4C Monte Carlo code met the requirements of TG43UI report. Dose rate constant, 0.977 78 cGy/(h×U), was in agreement within 1.32% compared with the recommended value of TG43UI. There was a good agreement between TG43UI about the dosimetric parameters at distances of 1 to 10 cm along the transverse axis of the 125I source established by MCNP4C and current published data. And the dose distribution of 125I photon emitting source in different bone tissue was calculated. Dose-deposition capacity of photons was in decreasing order: cortical bone, spongy bone, cartilage, yellow bone marrow, red bone marrow in the same medium depth. Photons deposited significantly in traversal axis among the phantom material of cortical bone and sponge bone relevant to the dose to water. In the medium depth of 0.01 cm, 0.1 cm, and 1 cm, the dose in the cortical bone was 12.90 times, 9.72 times, and 0.30 times of water respectively.@*CONCLUSION@#This study build a 125I source model with MCNP4C Monte Carlo code, which is validated, and could be used in subsequent study. Dose distribution of photons in different bone medium is not the same as water, and its main energy deposits in bone medium surface, so we should consider the effect of bone medium when we design the target area adjacent to the bone tissue in 125I sources implantation plan.

Prokaryotic expression,purification and identification of low-toxic staphylococcal enterotoxin A with D227A mutation / 中国免疫学杂志

Objective:To obtain recombinant D227A mutation Staphylococcal enterotoxin A protein(rSEAD227A) with low toxicity but still retain its immunological activity and high purity. Methods: The SEA gene containing D227A mutation was cloned by PCR. By constructing pET44a-SEAD227Avector and transfecting the expression strain Rosetta, inclusion bodies were solubilized with guanidium hydrochloride and refolded by gradient dialysis;proteins were purified using StrepⅡ affinity chromatography,and identified by Western blot and high performance liquid chromatography-mass spectrometry(LC-MS/MS). Results: The D227A mutation of SEA was cloned and the expression system of Rosetta-rSEAD227Awas constructed. The purified rSEAD227Aprotein was obtained by refolding with gradient dialysis and affinity purification. LC-MS/MS analysis confirmed that the tryptic digested rSEAD227A peptide sequences matched the sequences of SEA in the database. Conclusion: The rSEAD227A protein in high purity was obtained,which provided the ex-perimental basis for further basic research and clinical application of SEA.

Inductive effect of BuShenYangGuTang on apoptosis of multiple myeloma cell line KM3 and its mechanism / 天津医药

Objective To study the effects and mechanisms of BuShenYangGuTang on proliferation and apoptosis of multiple myeloma cell line KM3. Methods The inhibitory effect of BuShenYangGuTang on cell proliferation was assessed by CCK-8. BuShenYangGuTang induced KM3 cell cycle arrest was analyzed by flow cytometry after propidium iodide staining. Flow cytometry was also used to analyze the cell apoptosis after Annexin V-FITC staining. The expressions of Bcl-2, Bax and NF-κB were determined by RT-qPCR and Western blot assay. Results BuShenYangGuTang inhibited the KM3 cell proliferation in a dose-dependent manner. The expression levels of Bcl-2 and NF-κB were decreased, the expression level of Bax was increased, and the cell cycle was arrested in G0 / G1 phase after treatment with BuShenYangGuTang. Conclusion BuShenYangGuTang could inhibit the proliferation, arrest cell cycle and induce the apoptosis in KM3 cells, which may be related to the abnormal expressions of Bcl-2, Bax and NF-κB.

Modified poisoning severity score for early prognostic evaluation in acute paraquat poisoning / 解放军医学杂志

Objective To study the applied value of modified poisoning severity score (PSS) for early prognostic evaluation in acute paraquat poisoning.Methods Thirty-seven patients with acute paraquat poisoning from June 2013 to June 2016 were enrolled.The PSS score,the modified PSS score,the acute physiology and the chronic health status Ⅱ score (APACHE Ⅱ) of the patients were calculated.The relationship between modified PSS and APACHE Ⅱ was analyzed.Also the factors that affect outcome were analyzed by logistic regression analysis.The work characteristic curve (ROC curve) of the PSS,the modified PSS and the APECH Ⅱ were drawn and compared.Results There was a positive correlation between the risk of death and admission time,poisonous dose,the concentration of urine paraquat,and white blood cell count (P＜0.05).There was a significant correlation between the modified PSS and the APACHE Ⅱ (P＜0.0001).The immediate PSS score,the modified PSS score,and the APACHE Ⅱ score were significant for the prognosis of patients with acute paraquat poisoning.The area under the curve (AUC) was in turn 0.774,0.788,0.799.Among them,the best bound of the modified PSS score was 6.5 (when the score is greater than 6.5,the risk of death is higher).Further comparison of the area under the three curves showed that there was no significant difference in the area under the ROC curve between the three scores in predicting the prognosis of death [P=0.7633(PSS-DPSS),P=0.7791 (PSS-APACHE Ⅱ),P=0.8918(DPSS-APACHE Ⅱ)].Conclusion Modified PSS is helpful in early predicting the prognosis of acute paraquat poisoning.

Acute Fornix Deep Brain Stimulation Improves Hippocampal Glucose Metabolism in Aged Mice / 中华医学杂志(英文版)

<p><b>Background</b>A beneficial memory effect of acute fornix deep brain stimulation (DBS) has been reported in clinical studies. The aim of this study was to investigate the acute changes in glucose metabolism induced by fornix DBS.</p><p><b>Methods</b>First, the Morris water maze test and novel object recognition memory test were used to confirm declined memory in aged mice (C57BL/6, 20-22 months old). Then, four groups of mice were used as follows: aged mice with stimulation (n = 12), aged mice with sham-stimulation (n = 8), adult mice (3-4 months old) with stimulation (n = 12), and adult mice with sham-stimulation (n = 8). Ipsilateral hippocampal glucose metabolism and glutamate levels were measured in vivo by microdialysis before, during, and after fornix DBS treatment. Histological staining was used to verify the localization of electrodes and mice with inaccurate placement were excluded from subsequent analyses. The effects of fornix DBS on extracellular glucose, lactate, pyruvate, and glutamate levels over time were analyzed by repeated-measures analysis of variance followed by Fisher's least significant difference post hoc test.</p><p><b>Results</b>The aged mice had a higher basal lactate/pyruvate ratio (LPR) and lactate/glucose ratio (LGR) than the adult mice (LPR: 0.34 ± 0.04 vs. 0.13 ± 0.02, t = 4.626, P < 0.0001; LGR: 6.06 ± 0.59 vs. 4.14 ± 0.36, t = 2.823, P < 0.01). Fornix DBS decreased the ipsilateral hippocampal pyruvate and lactate levels (P < 0.05), but the glucose levels were not obviously changed in aged mice. Similarly, the LGR and LPR also decreased in aged mice after fornix DBS treatment (P < 0.05). Glucose metabolism in adult mice was not significantly influenced by fornix DBS. In addition, fornix DBS significantly decreased the ipsilateral hippocampal extracellular levels of glutamate in aged mice (P < 0.05), while significant alterations were not found in the adult mice.</p><p><b>Conclusions</b>The present study provides experimental evidence that fornix DBS could significantly improve hippocampal glucose metabolism in aged mice by promoting cellular aerobic respiration activity.</p>

Icaritin Reverses Multidrug Resistance of Multiple Myeloma Cell Line KM3/BTZ / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the reversing effect of icaritin on multidrug resistance of multiple myeloma cell lines KM3/BTZ and its underlying mechanism.</p><p><b>METHODS</b>KM3/BTZ cells were established by a gradually ascending gradient induction of bortezomib (BTZ). The sensitivities of KM3 and KM3/BTZ cells to 7 chemotherapeutic drugs, the inhibition and reversal effects of icaritin on proliferation and drug-resistance of KM3/BTZ cells were analyzed by MTT. The apoptosis was analyzed by flow cytometry, and the expression of Par-4, HSP27 and P-gp were detected by Western blot.</p><p><b>RESULTS</b>KM3/BTZ cells were not only resistant to BTZ, but also to other 6 chemotherapeutic drugs. The resistance index (RI) to BTZ was 17.84, and higher than that of other chemotherapeutic drugs. Icaritin inhibited the proliferation and induced the apoptosis of KM3/BTZ cells. The ICvalue of BTZ decreased from 0.345 µg/ml to 0.149 µg/ml, and the reversal index was 2.38 (P<0.05). The expression of Par-4 protein increased in a concentration-dependent manner, while the expression of HSP27 and P-gp were down-regulated.</p><p><b>CONCLUSION</b>Icaritin can inhibit cell proliferation and induce apoptosis of KM3/BTZ cells, moreover, can effectively reverse the multidrug resistance of KM3/BTZ cells. The mechanism may be related with down-regulation of HSP27 and P-gp expression, and up-regulation of Par-4 expression.</p>

Stimulation of Anterior Thalamic Nuclei Protects Against Seizures and Neuronal Apoptosis in Hippocampal CA3 Region of Kainic Acid-induced Epileptic Rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The antiepileptic effect of the anterior thalamic nuclei (ANT) stimulation has been demonstrated; however, its underlying mechanism remains unclear. The aim of this study was to investigate the effect of chronic ANT stimulation on hippocampal neuron loss and apoptosis.</p><p><b>METHODS</b>Sixty-four rats were divided into four groups: The control group, the kainic acid (KA) group, the sham-deep brain stimulation (DBS) group, and the DBS group. KA was used to induce epilepsy. Seizure count and latency to the first spontaneous seizures were calculated. Nissl staining was used to analyze hippocampal neuronal loss. Polymerase chain reaction and Western blotting were conducted to assess the expression of caspase-3 (Casp3), B-cell lymphoma-2 (Bcl2), and Bcl2-associated X protein (Bax) in the hippocampal CA3 region. One-way analysis of variance was used to determine the differences between the four groups.</p><p><b>RESULTS</b>The latency to the first spontaneous seizures in the DBS group was significantly longer than that in the KA group (27.50 ± 8.05 vs. 16.38 ± 7.25 days, P = 0.0005). The total seizure number in the DBS group was also significantly reduced (DBS vs. KA group: 11.75 ± 6.80 vs. 23.25 ± 7.72, P = 0.0002). Chronic ANT-DBS reduced neuronal loss in the hippocampal CA3 region (DBS vs. KA group: 23.58 ± 6.34 vs. 13.13 ± 4.00, P = 0.0012). After chronic DBS, the relative mRNA expression level of Casp3 was decreased (DBS vs. KA group: 1.18 ± 0.37 vs. 2.09 ± 0.46, P = 0.0003), and the relative mRNA expression level of Bcl2 was increased (DBS vs. KA group: 0.92 ± 0.21 vs. 0.48 ± 0.16, P = 0.0004). The protein expression levels of CASP3 (DBS vs. KA group: 1.25 ± 0.26 vs. 2.49 ± 0.38, P < 0.0001) and BAX (DBS vs. KA group: 1.57 ± 0.49 vs. 2.80 ± 0.63, P = 0.0012) both declined in the DBS group whereas the protein expression level of BCL2 (DBS vs. KA group: 0.78 ± 0.32 vs. 0.36 ± 0.17, P = 0.0086) increased in the DBS group.</p><p><b>CONCLUSIONS</b>This study demonstrated that chronic ANT stimulation could exert a neuroprotective effect on hippocampal neurons. This neuroprotective effect is likely to be mediated by the inhibition of apoptosis in the epileptic hippocampus.</p>

Behavioral effects of deep brain stimulation of the anterior nucleus of thalamus, entorhinal cortex and fornix in a rat model of Alzheimer's disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Recent clinical and preclinical studies have suggested that deep brain stimulation (DBS) can be used as a tool to enhance cognitive functions. The aim of the present study was to investigate the impact of DBS at three separate targets in the Papez circuit, including the anterior nucleus of thalamus (ANT), the entorhinal cortex (EC), and the fornix (FX), on cognitive behaviors in an Alzheimer's disease (AD) rat model.</p><p><b>METHODS</b>Forty-eight rats were subjected to an intrahippocampal injection of amyloid peptides 1-42 to induce an AD model. Rats were divided into six groups: DBS and sham DBS groups of ANT, EC, and FX. Spatial learning and memory were assessed by the Morris water maze (MWM). Recognition memory was investigated by the novel object recognition memory test (NORM). Locomotor and anxiety-related behaviors were detected by the open field test (OF). By using two-way analysis of variance (ANOVA), behavior differences between the six groups were analyzed.</p><p><b>RESULTS</b>In the MWM, the ANT, EC, and FX DBS groups performed differently in terms of the time spent in the platform zone (F(2,23) = 6.04, P < 0.01), the frequency of platform crossing (F(2,23) = 11.53, P < 0.001), and the percent time spent within the platform quadrant (F(2,23) = 6.29, P < 0.01). In the NORM, the EC and FX DBS groups spent more time with the novel object, although the ANT DBS group did not (F(2,23) = 10.03, P < 0.001). In the OF, all of the groups showed a similar total distance moved (F (1,42) = 1.14, P = 0.29) and relative time spent in the center (F(2,42) = 0.56, P = 0.58).</p><p><b>CONCLUSIONS</b>Our results demonstrated that DBS of the EC and FX facilitated hippocampus-dependent spatial memory more prominently than ANT DBS. In addition, hippocampus-independent recognition memory was enhanced by EC and FX DBS. None of the targets showed side-effects of anxiety or locomotor behaviors.</p>