Aim To investigate the effect of p38 MAPK AS-ODNs on the expression of GLT-1 during the induction of brain ischemic tolerance induced by cerebral ischemic preconditioning (CIP).Methods Eighty healthy male Wistar rats with permanent occlusion of the bilateral vertebral arteries were randomly divided into 6 groups: ①Sham group (n=10);②CIP group (n=10);③ischemic insult (Ⅱ) group (n=10);④CIP+Ⅱ group (n=10);⑤p38 MAPK AS-ODNs+CIP+Ⅱ group (n=30);⑥p38 MAPK S-ODNs+CIP+Ⅱ group (n=10).Group ⑤ was divided into 5 nmol, 10 nmol and 15 nmol subgroups according to the dose of p38 MAPK AS-ODNs (n=10).The dose of p38 MAPK S-ODNs was 15 nmol.All the rats were sacrificed 6 h and 2 d after the sham operation or the last time of global brain ischemia reperfusion.Western blot and immunohistochemistry analysis were used for detecting the expression of p-p38 MAPK and GLT-1 protein.Results CIP moderately up-regulated the expression of p-p38 MAPK and significantly up-regulated the expression of GLT-1 protein, inhibited the excessively up-regulation of p-p38 MAPK and the down-regulation of GLT-1 induced by ischemic insult.p38 MAPK AS-ODNs significantly inhibited the up-regulation of p-p38 MAPK and GLT-1 protein in a dose-dependent manner during the induction of brain ischemic tolerance by CIP.Conclusion p38 MAPK AS-ODNs inhibit the up-regulation of GLT-1 during the induction of brain ischemic tolerance induced by CIP.

To prepare the RNAse-resistant virus particles containing the partial gene fragments of avian influenza virus H5N1 for use as RNA standard and control in RNA virus detection, the genes coding the coat protein and maturase of E.coli bacteriophage MS2 were amplified by PCR and then cloned into prokaryotic expression vector pET32a to construct the intermediate vector pET32a-MS2. In addition, the gene sequences coding hemagglutinin (HA), neuraminidase(NA) and M protein of the H5N1 virus were also cloned separately to the down-stream of plasmid pET32a-MS2, thus constructing the prokaryotic expression vectors pET32a-NS2-HA, pET32a-MS2-NA and pET32a-MS2-M. These recombinant plasmids were then transformed separately to E.coli BL21(DE3) with induction by IPTG. to express the virus-like particles. The virus-like particles observed under electron microscopy were identified by RT-PCR ,while their stability was confirmed by real-time RT-PCR. In this way, the virus-like particles were successively constructed and identified through PCR amplification, enzymolysis identification and sequencing analysis. These virus-like particles observed under electron microscopy appeared to be circular in shape with a diameter of about 50 nm. Their stability was proved to be rather good. From these observations, it is apparent that these virus-like particles can be used as RNA standard and quality control in the detection of avian influenza virus H5N1.

Objective:To analyze the magnetic resonance imaging (MRI) features of diffuse midline gliomas with H3K27M mutation, and to quantitatively analyze the changes of apparent diffusion coefficient (ADC).Methods:The MRI images of 14 cases of diffuse midline gliomas with H3K27M mutation were retrospectively analyzed in the Affiliated Hospital of Qingdao University from April 2017 to November 2019. The location, edge, signal, peritumoral edema and enhancement characteristics of the lesions were observed, and the changes of ADC values were analyzed.Results:The tumors were located in thalamus in four cases, pons in six cases, medulla oblongata in two cases and spinal cord in two cases. In seven cases, the tumor was confined to the midline region, of which six cases had clear boundary, seven cases were located in the midline area, but infiltrated into the non midline area at the same time, and six cases had unclear boundary. Basilar artery entrapment was found in all six patients located in pons. Multiple large cystic changes were found in five cases, multiple small cysts in four cases and no cystic changes in five cases. Cystic changes were found in all seven cases of tumors involving the non midline region, of which six cases were located only in the non midline region, and only two of the seven tumors localized in the midline region had small cysts. Hemorrhage was found in four cases. Five cases showed mild heterogeneous enhancement, six cases showed moderate heterogeneous enhancement, two cases showed obvious enhancement, and one case showed no enhancement. There was no edema around the tumor in nine cases and mild edema in five cases. The average edema index was 1.13. The average ADC value of tumor parenchyma in 12 patients was (7.83±0.88)×10 -4 mm 2/s, which was 15.6% lower than that of the contralateral side [(9.28±0.69)×10 -4 mm 2/s, t=-6.336, P<0.05]. Conclusions:Diffuse midline gliomas with H3K27M mutation have a younger onset age and are more likely to occur in thalamus, brainstem and spinal cord. Most of the tumors have no peritumoral edema or mild peritumoral edema. The tumors confined to the midline region are regular in shape and clear in boundary. The masses involving the non midline area are prone to cystic necrosis. Diffuse midline gliomas with H3K27M mutation in pons are prone to basilar artery entrapment. ADC value can provide a quantitative basis for preoperative tumor grading.

Objective:To investigate the effect of vector sum concept in fine-tuning posterior column screw channel via ilioinguinal approach for the treatment of bi-column acetabular fracture.Methods:A retrospective cohort study was conducted to analyze the clinical data of 42 patients with acetabular double column fracture admitted to Weifang People′s Hospital from July 2015 to May 2021, including 22 males and 20 females, aged 23-77 years [(49.3±16.3)years]. The ilioinguinal approach was used in all patients with the anterior column fixed with a plate and posterior column fixed with a lag screw. The vector sum concept was used intraoperatively to fine-tune the posterior column screw channel in 19 patients (channel fine-tuning group): namely, a 2.5 mm Kirschner wire was inserted into the bony channel of the posterior column screw under fluoroscopy of iliac oblique and obturator oblique positions; when the Kirschner wire was not located in the middle of the ischial ramus under single fluoroscopy, the vector only needed to be adjusted in one direction, with zero in the other direction; when the Kirschner wire was not located in the middle of the ischial ramus under fluoroscopy of both the iliac oblique and obturator obturator oblique positions, the sum of the deviation vectors in the two directions was calculated before fine-tuning. The vector sum concept was not used to fine-tune the posterior column channel screw in 23 patients (channel non-fine-tuning group). The time of posterior column screw placement, intraoperative blood loss, frequency of guide wire adjustment and fracture healing time were recorded and compared between the two groups. At 6 months after operation, the quality of fracture reduction and hip function were assessed by Matta score and Merle D′Aubigne-Postel score, respectively. The complications were observed.Results:All patients were followed up for 7-71 months [(35.7±8.5)months]. In channel fine-tuning group, the time of posterior column screw placement was (5.1±1.5)minutes, with intraoperative blood loss of (798.8±83.9)ml, frequency of guide wire adjustment of (1.8±0.5)times and fracture healing time of (12.4±3.2)weeks; while these parameters [(39.8±12.0)minutes, (1 119.3±172.0)ml, (5.6±1.6)times and (15.6±4.2)weeks] were significantly shorter or less in channel non-fine-tuning group ( P<0.05 or 0.01). There were no significant difference in the quality of fracture reduction and hip function between the two groups at 6 months postoperatively (all P>0.05). After operation, symptoms of lateral femoral cutaneous nerve was found in seven patients, superficial incision infection in two who was healed after debridement and dressing change, deep venous thrombosis of lower limbs in three. There was no significant difference in the incidence of postoperative complications between the two groups [channel fine-tuning group: 26%(5/19), channel non-fine-tuning group: 30%(7/23)] ( P>0.05). Conclusion:For bi-column acetabular fractures via ilioinguinal approach, application of vector and concept to fine-tune the posterior column screw channel is beneficial for rapid screw placement into the osseous channel, significant reduction of intraoperative blood loss and early fracture healing.

Objective To compare the safety and efficacy of stent-assisted coil and non-stent-assisted coil for the treatment of ruptured posterior communicating artery aneurysms.Methods A total of 121 consecutive patients with ruptured posterior communicating artery aneurysm treated at the Department of Neurosurgery,Heze Municipal Hospital between June 2014 and June 2017 were enrolled retrospectively.They divided into either a stent group (n =63) or non-stent group (n =58) according to whether they used stent treatment or not.The two groups were compared,and their clinical data,surgical related complications,the degree of embolism immediately after aneurysm surgery,and the follow-up results of clinical and imaging were analyzed.Results Interventional embolization therapy was successfully performed in all the patients.(1) The proportion of wide-necked aneurysms of the stent group was higher than that of the non-stent group.There was significant difference (92.1％ [58/63] vs.8.6％ [5/58],x2 =84.249,P ＜ 0.01).There were no significant differences in age,aneurysm size,sex,and HuntHess grade (all P ＞ 0.05).(2) The incidences of intraoperative aneurysm rupture of the stent group and non-stent group were 4.8％ (3/63) and 3.4％ (2/58) respectively.There was no significant difference (x2 =0.132,P =0.717).The incidence of thromboembolie complication was 12.7％ (8/63) and 5.2％ (3/58)respectively.There was no significant difference (x2 =2.070,P =0.150).In the stent group,1 patient disabled and 1 died,and in the non-stent group,there were no cases with surgical related disability,and 1 died.(3) The comparison of immediate results after procedure between the stent group and non-stent group:there were no significant differences in the complete embolic rate (39.7％ [25/63] vs.37.9％[22/58],x2 =0.039),aneurysm neck residual rate (25.4％ [16/63] vs.39.7％ [23/58],x2 =2.811],and residual rate of aneurysm (34.9％ [22/63] vs.22.4％ [13/58],x2 =2.298,all P ＞ 0.05).(4) The recurrence rate of aneurysms (4.3％ [2/46]) of the stent group was lower than that of the non-stent group (31.8％ [14/44]).There was significant difference between the two groups (x2 =112.610,P =0.01).No rebleeding occurred in all aneurysms after procedure.Conclusion Compared with the non-stent-assisted coil embolization,the stent assisted embolization of ruptured posterior communicating artery aneurysms in acute phase may reduce the recurrence rate of aneurysms and without significantly increasing the incidence of surgery-related complications.

Objective:To establish a sequencing method for the genome of severe fever with thrombocytopenia syndrome virus (SFTSV) based on next-generation sequencing (NGS).Methods:SFTSV RNA was extracted from serum samples of patients with severe fever with thrombocytopenia syndrome. SFTSV-specific primers were designed using Primer 5.0 software. A multiplex PCR method was constructed and used to amplify the nucleotide sequence of SFTSV. Whole-genome sequencing was performed on the NGS platform.Results:The whole genes of SFTSV isolates in 28 serum samples were amplified by the multiplex PCR with a coverage over 94%. Sequencing and phylogenetic analysis of those strains revealed that the predominant strains ( n=20) belonged to genotype A, followed by genotypes B ( n=4) and E ( n=3). Conclusions:A high-throughput sequencing method for SFTSV based on multiplex PCR was established in this study. This method was characterized by high specificity and good quality and could improve the sequencing efficiency.

Objective:To perform a prospective cohort study to identify individual susceptibility of glucocorticoid (GC) -associated osteonecrosis of the femoral head (GA-ONFH) and their clinical and genetic risk factors. Methods:The present prospective cohort study enrolled patients who received their first GC therapy between July 2015 and January 2018 at Zhongshan Hospital. All patients did not receive any GC treatment before enrollment. Further, they planned to start GC treatment with the dose (equivalent prednisone) of ≥30 mg/d, lasted ≥3 weeks, or pulse dose ≥200 mg/d, lasted ≥3 d. Blood samples were collected before GC treatment to evaluate bone metabolism and its released factors. Hip MRI was performed at the 1st, 3rd, 6th, 12th and 24th month to diagnose GA-ONFH. All patients were followed-up for ≥2 years. The endpoint was regarded as diagnosis of GA-ONFH or completion of 2 years follow-up. Lasso regression was performed to determine which clinical features were associated with GA-ONFH. A nested case-control sub-cohort (A, n=12) was established prospectively based on the main cohort by 1∶1 matching. Whole exome sequencing was performed to screen differential and functional candidate single nucleotide polymorphisms and insertion-deletions (SNP/InDels). Another sub-cohort (B, n=50) was constructed retrospectively in patients with GA-ONFH and non-ONFH patients received standard high dose GC treatment for more than two years. The candidate SNP/InDels were verified by Sanger sequencing based on the patients from sub-cohort B. Results:A total of 96 patients were enrolled of which 88 of them (32 males and 56 females, mean age 42.30 years) completed follow-up. Eight cases (9.1%) were diagnosed with GA-ONFH. The median time from the start of GC therapy to the diagnosis of ONFH was 53.00(34.00,13.50) days. The baseline characteristics, such as age, sex and body mass index, indicated no significant difference between the ONFH group and the non-ONFH group. The cumulative GC dose of the ONFH patients in the first month was higher than that of non-ONFH [32.74(29.55, 47.05) mg/kg vs. 24.00(21.10, 29.45) mg/kg, Z=-2.410, P=0.016]. However, there was no significant difference of patients who underwent pulse therapy (37.5% vs. 10.0%, adjusted χ 2=2.829, P=0.093). The ratio of serum apolipoprotein B/apolipoprotein A1 (ApoB/ApoA1) in patients with ONFH was higher than that in non-ONFH group before GC use [0.95(0.80, 1.50) vs. 0.70(0.60, 0.80), Z=-2.875, P=0.000]. Due to the multicollinearity, Lasso regression model was performed to reduce overfitting. All variables were included in the model. The results suggested that higher ApoB/ApoA1 ratio, lower serum β-c-terminal telopeptide (β-CTX) and higher cumulative GC dose in the first month were the top three risk factors of GA-ONFH. This model had an accuracy of 0.982 in internal validation. Seven differential candidate SNP/InDels were found by whole exome sequencing of sub-cohort A. We further verified these SNP/InDels in sub-cohort B. The patients with COLEC12 mutation (rs2305027, G1816A) were at risk of GA-ONFH ( OR=6.00, 95% CI: 1.17, 30.73). Conclusion:Higher first-month GC dose, lower serum β-CTX level before treatment, higher ApoB/ApoA1 ratio and COLEC12 mutation (rs2305027, G1816A) could increase the risk of GA-ONFH.

Objective:To report the first case of sever fever with thrombocytopenia syndrome caused by severe fever with thrombocytopenia syndrome virus (SFTSV) in Puyang city, and to study the epidemiological and molecular characteristics of S, M, L fragments of the SFTSV isolate.Methods:The epidemiological characteristics of this case was analyzed with epidemiological methods. SFTSV was isolated from the patient′s serum sample. Nucleic acid of SFTSV was extracted and detected by fluorescent RT-PCR. A multiplex PCR method was constructed to amplify the nucleic acid sequence of the virus. whole-genome sequencing was performed on the next-generation sequencing platform. MEGA11 and DNAStar was used for homology analysis and a phylogenetic tree was constructed.Results:Epidemiological investigation showed that the patient and his close contacts had no history of travel or tick bite within 14 d, but had a history of fieldwork. The patient′s serum sample was positive for SFTSV nucleic acid. Genetic analysis showed that the S, M, L gene fragments of the first SFTSV isolate in Puyang belonged to genotype E. This isolate shared 94.8%-99.6%, 94.0%-99.8% and 95.7%-99.7% nucleotide sequence homology with the representative strains acquired from GeneBank in S, M, L gene fragments, respectively.Conclusions:This case was the first case of SFTSV-caused severe fever with thrombocytopenia syndrome in Puyang. The SFTSV isolate shared a close homology with domestic isolates, but its genotype was significantly different from the SFTSV strains isolated in Henan in recent years, indicating that it might an imported case from other places in Henan Province or Hubei Province. Disease monitoring and professional training for medical personnel should be strengthened and more attention should be paid to the evolution and mutation of SFTSV.

Objective:To analyze the epidemiological characteristics and spatiotemporal characteristics of human brucellosis in Henan Province.Methods:Data of human brucellosis in Henan Province from 2005 to 2021 were collected through the China Disease Prevention and Control Information System, and a descriptive epidemiological method was used to analyze the epidemic profile of brucellosis in Henan Province and the three distribution characteristics. Global and local spatial autocorrelation were used to analyze the spatial distribution and the hot spots of brucellosis in Henan Province, respectively, and spatiotemporal scanning was used to analyze the spatiotemporal clustering regions of brucellosis in Henan Province.Results:A total of 39 862 brucellosis cases were reported in Henan Province from 2005 to 2021, with an average annual incidence of 2.44/100 000, and the number of cases showed an overall increasing trend each year (χ 2trend = 11 127.85, P = 0.001). The onset months were mainly concentrated from March to July, accounting for 59.00% (23 517/39 862), with May as the peak (5 478 cases). Cases of brucellosis were reported in 157 counties (cities, districts) of the province. The ratio of male to female was 2.52∶1.00 (28 542/11 320). Farmers were the main occupation, with 32 985 cases (82.75%). The age of onset was mainly 45 to 65 years old, with 20 226 cases (50.74%). The global spatial autocorrelation analysis showed that the global Moran's I was > 0, Z > 1.96, and P < 0.05 in all years except 2006 - 2008, showing spatial clustering. Further local spatial autocorrelation analysis was performed, and high-high and low-low clustering areas existed in 2012 - 2021 ( P < 0.01). Spatiotemporal scanning analysis showed that there was one spatiotemporal cluster in the high incidence area and two spatiotemporal clusters in the low incidence area. The high incidence cluster was centered in Neixiang County, covering 48 counties (cities, districts) including Song County and Ruzhou City, and the aggregation time was from 2014 to 2021. The two low incidence clusters were centered in Yongcheng City and Boai County, covering 58 and 18 counties (cities, districts), respectively, and the aggregation time was 2016 - 2021 and 2005 - 2012, respectively. Conclusion:The overall incidence of brucellosis in Henan Province is on the rise from 2005 to 2021, with middle-aged and elderly male farmers as the main affected population, and there are spatiotemporal clusters of brucellosis in Henan Province.

Diabetes retinopathy （DR） is an important cause that threatens the visual health of adults. There are some treatment methods of western medicine with definite efficacy， such as anti-vascular endothelial growth factor and laser photocoagulation， but they have many adverse reactions such as intraocular infection and visual field damage. Traditional Chinese medicine （TCM） therapies are safe and effective， which can complement western medicine. Phosphatidylinositol3-kinase （PI3K）/protein kinase B （Akt） signaling pathway regulates a range of processes including glucose metabolism， cell proliferation， and cell transcription and apoptosis， which is closely related to the occurrence and development of DR. Numerous studies have shown that TCM monomers can participate in maintaining the integrity of blood-retinal barrier and inhibiting retinal neovascularization and neurodegeneration in many aspects such as inhibiting oxidative stress and alleviating inflammatory reaction by regulating the PI3K/Akt pathway， so as to delay the progress of DR. Therefore， this study reviewed PI3K/Akt pathway and its relationship with DR， as well as the TCM monomers in interfering with DR based on PI3K/Akt pathway to provide some ideas for the prevention and treatment of DR in integrated TCM and western medicine.