[Research progress on Mycobacterium tuberculosis acetyltransferase].

The protein acetylation of Mycobacterium tuberculosis(MTB) plays an important role in virulence, drug resistance, regulation of metabolism and host anti-tuberculosis immune response. The proteins acetylation of MTB and host protein could be induced by the MTB acetyltransferase, which is related to the occurrence, development and prognosis of tuberculosis (TB). A clear understanding of the function of MTB acetyltransferase and identification of its targeted regulatory protein acetylation modification is critical to elucidate the pathogenic mechanism and drug resistance mechanism of TB, and then this could then provide new targets for the development of anti-tuberculosis drugs. This article systematically reviewed the research progress on MTB acetyltransferase related functions, which will provide a theoretical basis for further research on its mediated protein acetylation modification, further development of new anti-tuberculosis drugs and elucidation of drug resistance mechanism.

[A case of von Hippel-Lindau syndrome with optic disc pit and macular hole].

A 69-year-old female patient presented to the ophthalmology department with complaints of blurred vision in the left eye for more than 10 days. Her medical history revealed a history of right renal tumor and left pheochromocytoma, which were treated with surgical resection at an external institution. Ophthalmic examination revealed a temporal superior cup-shaped optic disc pit in the left eye, along with a macular hole approximately 1/5 the size of the optic disc diameter in the macular region. Additionally, peripheral retinal examination at the 6 o'clock and 11 o'clock positions showed vascular tumors, each approximately 1.5 times the size of the optic disc diameter. Based on the patient's medical history, fundus findings, and auxiliary examination results, a diagnosis of macular hole in the left eye, optic disc pit in the left eye, and Von Hippel-Lindau (VHL) syndrome was established. Subsequently, the patient underwent left vitrectomy and macular hole repair surgery, leading to an improvement in visual acuity.

[Research progress in non-coding RNAs in mediating immune responses of macrophages to Mycobacterium tuberculosis infection].

Tuberculosis (TB) is a chronic infectious disease caused by mycobacterium tuberculosis (MTB) infection. Macrophages are the first line in defensing MTB infection and the main host cells for the growth and persistence of MTB. Changes in macrophage function are critical for the host response to tuberculosis. Non-coding RNAs are involved in the pathophysiological process of many diseases, including TB, and play a very important regulatory role in the macrophage mediated immune response process. Therefore, we reviewed the mechanisms of the non-coding RNAs mediated function alteration of macrophages, in order to facilitate identification of potential therapeutic targets for host-directed anti-TB treatment.

[Mechanism of Mycobacterium tuberculosis on interleukin-6 receptor 3'-untranslated region methylation in CD4+T cells].

Objective: To investigate the role and mechanism of DNA methylation in Mycobacterium tuberculosis (MTB lysate) -induced downregulation of interleukin-6 receptor(IL-6R) expression in CD4+T cells. Methods: A prospective study was conducted. Bisulfite sequencing (BSP) was applied to determine the methylation levels of CpG island in IL-6R promoter region and 3'untranslated region (3'UTR) region in CD4+T cells from peripheral blood mononuclear cells (PBMC) of control group (healthy person, n=10) and TB group (tuberculosis patients, n=10) in Shenzhen Third People's Hospital between 2019 and 2020. Quantitative reverse transcription-PCR (RT-qPCR) and Western blotting were used to detect the expression of IL-6R, DNMT1, DNMT3A and DNMT3B in MTB lysate-stimulated CD4+T cells and Jurkat E6-1 cells. Furthermore, PBMC in control group and Jurkat E6-1 cells activated by anti-CD3/CD28 antibody were stimulated by MTB lysates to detect the methylation levels of CpG island and IL-6R and DNMT expression. Transcriptional activity of differently methylation regions of IL-6R 3'UTR was detected by using luciferase reporter gene system. Results: IL-6R expression in TB group was lower than that in control group, but DNMT1 and DNMT3B expressions were higher than those in control group in CD4+T cells isolated from PBMC. There was no significant difference in the methylation rate of IL-6R promoter CpG island of CD4+T cells between control and TB group. However, the methylation rates of CpG island in 3'UTR region were significantly higher (P<0.001) in TB (69.5%±3.4%), compared with control (54.3%±4.7%). Besides, IL-6R expression was lower than unstimulated, while DNMT1 and DNMT3B expression was higher than unstimulated after MTB lysate-stimulation of activated control PBMC in vitro. The methylation rate of CpG island in IL-6R 3'UTR region of CD4+T cells increased from 58.8%±11.6% to 79.4%±10.9% (P<0.001) after MTB lysate-stimulated PBMC of the control. The same results were observed in the MTB lysate-stimulated CD4+T cells isolated from PBMC in control and Jurkat E6-1 cell line. Furthermore, IL-6R expression after co-treatment of the DNA methyltransferase inhibitor decitabine (5-aza) with MTB lysate was higher than that stimulated by MTB lysate alone. In addition, the methylation levels of CpG islands in the 3' UTR region of IL-6R were lower than those stimulated by MTB lysates alone after co-treatment of the DNA methyltransferase inhibitor decitabine (5-aza) with MTB lysates. The transcriptional activity of the fully unmethylated IL-6R 3'UTR CpG island reporter gene was higher than that of the fully methylated IL-6R 3'UTR CpG island. Conclusions: MTB lysates stimulation inhibited IL-6R expression transcriptionalely as well as on the protein level by inducing hypermethylation of CpG island in IL-6R 3'UTR region of CD4+T cells. The hypermethylation of CpG island in IL-6R 3'UTR region of CD4+T cells induced by MTB may be related to the increased expression of DNMT1 and DNMT3B.

[The clinical value of further accurate staging of pT2 gastric cancer based on the depth of invasion].

Objective: To investigate the clinical value of pT2 gastric cancer staging pT2a and pT2b according to the depth of muscularis propria invasion in evaluating the prognosis of gastric cancer. Methods: According to the 8th edition of TNM staging system for gastric cancer proposed by the Union for International Cancer Control (UICC) and the American Joint Committee on Cancer (AJCC), patients with gastric cancer who underwent radical surgery in the Fourth Hospital of Hebei Medical University from January 1, 2008 to January 1, 2015 were selected and divided into pT2a and pT2b stage group according to the depth of tumor invasion. The 5-year overall survival (OS) and disease-free survival (DFS) were compared between the two groups. Results: The median follow-up time of 1 411 patients with postoperative pathological pT2 stage was 68.8 months, and 1 347 patients (95.46%) received complete follow-up data. The 5-year OS rate was 65.85%, and the 5-year DFS rate was 67.83 %. The 5-year OS rate and 5-year DFS rate of 709 pT2a patients were 72.50% and 73.91%, respectively. The 5-year OS rate and 5-year DFS rate of 638 pT2b patients were 58.46% and 61.13%, respectively, significantly different from those of the pT2a group (P<0.001). Hierarchical analysis was performed according to N staging. The 5-year OS rates of pT2aN0M0 (274 cases), pT2aN1M0 (192 cases), pT2aN2M0 (147 cases), pT2aN3aM0 (59 cases) and pT2aN3bM0 (37 cases) were 83.58 %, 72.40 %, 68.71 %, 54.24 % and 35.12 %, respectively. The 5-year DFS rates were 84.67 %, 77.08 %, 67.35 %, 54.24 % and 35.14 %, respectively. In the pT2b group, the 5-year OS rates of pT2bN0M0 (209 cases), pT2bN1M0 (166 cases), pT2bN2M0 (127 cases), pT2bN3aM0 (78 cases) and pT2bN3bM0 (58 cases) were 76.08%, 62.05%, 56.69%, 37.18% and 17.24%, respectively, and the 5-year DFS rates were 80.86%, 69.28%, 54.33%, 35.90% and 15.52%, respectively. Under the same N stage, the OS rates of patients in the pT2a group were better than those in the pT2b group (P values were 0.023, 0.034, 0.034, 0.043 and 0.018, respectively). When the N stage was N0 and N1, there was no significant difference in the 5-year DFS rate between the pT2a group and the pT2b group (P values were 0.199 and 0.090, respectively). When the N stages were N2, N3a and N3b, the difference between the pT2a stage group and the pT2b stage group was statistically significant (P values were 0.027, 0.022 and 0.025, respectively). Conclusions: In the 8th edition of AJCC/UICC gastric cancer staging system, pT2 stage can be divided into pT2a stage (invasion of superficial muscularis) and pT2b stage (invasion of deep muscularis) according to the infiltration depth of muscularis propria. There are significant differences in prognosis between the two groups. Combined with the number of lymph node metastasis, the prognosis of patients with pT2 gastric cancer can be more accurately evaluated.

[The effect of cancer nodules on survival prognosis of gastric cancer patients].

Objective: To explore the relationship between cancer nodules and clinicopathological characteristics of gastric cancer, and analyze its impact on survival prognosis of gastric cancer patients. Methods: A retrospective analysis of 2 386 patients with gastric cancer who underwent radical surgery from January 1, 2012 to January 1, 2015 in the Third Surgery Department of the Fourth Hospital of Hebei Medical University was performed. The relationship between cancer nodules and clinicopathological characteristics of gastric cancer and its impact on survival prognosis of gastric cancer patients were analyzed. Results: Among the 2 386 patients, there were 459 cases (19.24%) with cancer nodules, and 1 927 cases (80.76%) without cancer nodules. Logistic multivariate analysis showed that pT staging (P=0.036), pN staging (P=0.024), pTNM staging (P=0.032), Borrmann classification (P=0.008), vascular tumor thrombus (P=0.001) were independent risk factors for cancer nodules. The complete follow-up date of 2 273 cases (95.26%) of 2 386 patients with gastric cancer were obtained. A total of 1 259 patients relapsed and 1 152 died during the follow-up period. The 5-years overall survival (OS) rate was 49.32%, and the 5-years disease-free survival (DFS) rate was 44.61%. Among them, the 5-years OS rate and DFS rate of those with cancer nodules were 26.76% and 24.94%, while the 5-years OS rate and DFS rate of those without cancer nodules were 54.75% and 49.34%, respectively (P<0.001). Patients with positive cancer nodules were divided into 3 groups according to the number of cancer nodules: 1 (115 cases), 2 to 3 (202 cases), and more than 4 (124 cases). The 5-years OS rates of 3 groups were 41.74%, 30.69% and 10.48%, respectively (P<0.001). The 5-years DFS rates were 40.00%, 28.22% and 9.68%, respectively (P<0.001). Cox multivariate analysis showed that histological type (P=0.004), pT staging (P=0.007), pN staging (P=0.004), pTNM staging (P=0.002), vascular tumor thrombus (P=0.034), cancer nodules (P=0.005) and the number of cancer nodules (P=0.001) were independent risk factors for the prognosis of gastric cancer patients, and postoperative adjuvant chemotherapy (P=0.043) was a protective factor for the prognosis of gastric cancer patients. Conclusion: Cancer nodules are closely related to the tumor stage and prognosis of gastric cancer patients. The number of cancerous nodules is an independent risk factor for the prognosis of gastric cancer patients.

[Diagnostic performance of a novel Mycobacterium Tuberculosis specific T-Cell based assay for tuberculosis].

Objective: To evaluate the diagnosic performance of a novel Mycobacterium tuberculosis (MTB) specific T-cell based assay for tuberculosis, which targets the mRNA detection of interferon gamma-induced protein 10 (IP-10). Methods: Suspected tuberculosis patients were prospectively and consecutively recruited in Beijing Chest Hospital between March 2018 and November 2019, and individuals with lower risk of MTB infection were also recruited. IP-10.TB and T-SPOT.TB assays were simulataneously performed on peripheral blood samples. The diagnostic performance of IP-10.TB and T-SPOT.TB were analyzed using the receiver operating characteristic curve. Accordance of IP-10.TB and T-SPOT.TB was analyzed by Cohen's kappa test, while the correlation between the expression level of IP-10 mRNA in IP-10.TB test and the number of SFCs in T-SPOT.TB test were analyzed by Pearson correlation test. Results: A total of 235 patients with tuberculosis, 110 patients with other diseases and 153 individuals with lower risk of MTB infection were included in the final analysis. No significant difference was detected in the rate of indeterminate results between IP-10.TB assay (3/498, 0.60%) and T-SPOT.TB assay (6/498, 1.21%). The total sensitivity and specificity of IP-10.TB assay were 91.3% (95%CI 86.8%-94.6%) and 81.1% (95%CI 75.8%-85.7%). The specificity of IP-10.TB in individuals with lower risk of MTB infection was 98.0% (95%CI 94.4%-99.6%). The total sensitivity and specificity of T-SPOT.TB assay were 93.0% (95%CI 88.9%-96.0%) and 83.8% (95%CI 78.7%-88.1%). The specificity of T-SPOT.TB in individuals with lower risk of MTB infection was 100% (95%CI 97.6%-100.0%). No significant differences were detected in sensitivity and specificity between IP-10.TB and T-SPOT.TB assays (P>0.05). The positive coincidence rate of these 2 methods was 91.0% (95%CI 87.5%-94.5%), and the negative coincidence rate was 88.9% (95%CI 84.9%-92.9%) and the total coincidence rate was 90.0% (95%CI 87.3%-92.6%). The Cohen's kappa value was 0.80 (95%CI 0.75-0.85, P<0.001) between IP-10.TB and T-SPOT.TB assays. Conclusion: These results showed that the diagnostic performance of IP-10.TB was consistent with that in T-SPOT.TB, and this test could be a novel adjunctive tool for the diagnosis of tuberculosis.

[Pathological characteristics and survival analysis of 355 patients with gastroenteropancreatic neuroendocrine neoplasms].

Objective: Biological behavior, pathological characteristics and prognostic factors of 355 cases with gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs) were analyzed in this retrospective study. Methods: In our study, 355 patients pathologically diagnosed as GEP-NENs were identified from April 2006 to November 2017 in the Fourth Hospital of Hebei Medical University. The biological behavior, pathological characteristics and prognosis were analyzed retrospectively. Results: There were 355 patients (228 males and 127 females) with a mean age of 58.3±10.7 years. GEP-NENs were detected most frequently in the stomach (48.2%), followed by the pancreas (16.1%), colorectum (14.1%), esophagus (7.6%), duodenum/jejunum(5.6%), liver (4.2%), appendix (2.3%) and gallbladder/bile duct (2.0%). The main clinical manifestations of non-functional GEP-NENs were abdominal pain (88/350, 25.14%), ventosity (77/350, 22.00%) and dysphagia (68/350, 19.43%), which were generally lacking specificity at the first diagnosis. 295 patients were treated surgically, including 45 cases of endoscopic resection and 250 cases of laparoscopic operation. Concerning to pathological grading, there were 22.5% (80/355) patients in grade 1 (G1), 12.7% (45/355) in grade 2 (G2), and 58.9% (209/355) in grade 3 (G3). The median follow-up time was 34 months. Furthermore, the 1-, 3- and 5-year overall survival calculated by Kaplan-Meier method were 80.1%, 59.8%, and 57.5%, respectively. Univariate analysis revealed that tumor site, treatment, operation type, depth of tumor invasion, TNM staging, pathological grading, vascular embolus, lymph node metastasis, tumor size, preoperative leukomonocyte level and preoperative plasma albumin were associated with overall survival (all P<0.05). Multivariate analysis showed that treatment, operation type, depth of tumor invasion, TNM staging, pathological grading, vascular embolus, lymph node metastasis and tumor size were independent prognostic factors for GEP-NENs (all P<0.05). Conclusions: The clinicopathological characteristics of GEP-NENs should be mastered by clinicians, and the standard treatment measures were also needed to be formulated based on the prognostic factors in order to improve the prognosis of patients.

[Application of the pathological classification of "CCCG-WT-2016" (2019 revision) for treatment of Wilms tumors].

Objective: To describe our experiences in application of the 2019 revision of "CCCG-WT-2016" for the diagnosis of Wilms tumors. Methods: Ninety-one cases of Wilms tumor diagnosed at Shanghai Children's Medical Center from January 2015 to December 2018 were collected. All cases were reviewed by two senior pathologists, including one from China and the other from Singapore, according to the 2019 revision of "CCCG-WT-2016." Results: The specimens were obtained by core biopsy (n=21), primary nephrectomy (n=41), post-chemotherapy nephrectomy/resection (n=18), or biopsy/resection of metastatic/relapse/post-chemotherapy metastatic lesion(s) (n=11). The specimens of core biopsy and primary nephrectomy (n=62) all had favorable histology.Twelve post-chemotherapy nephrectomy cases were subdivided into three risk groups: low risk (n=0), intermediate risk (n=10) and high risk (n=2). Six post-chemotherapy resection cases were subdivided into 3 risk groups:low risk (n=0), intermediate risk (n=5) and high risk (n=1). The remaining 11 cases were comprised of metastatic, relapse, and post-chemotherapy metastatic lesions. The concordance rate of the two senior pathologists was 100%(91/91). Conclusions: The 2019 revision of "CCCG-WT-2016" is clearly written and easy to use. It can serve as the basis of accurate classification for clinical treatment.

[Clinicopathological characteristics of NTRK-rearranged mesenchymal tumors in childhood].

Objective: To investigate the clinical and pathological features of pediatric NTRK-rearranged tumors. Methods: Four NTRK-rearranged soft tissue tumors and one renal tumor at Shanghai Children's Medical Center, Shanghai Jiaotong University and Singapore KK Women's and Children's Hospital from January 2017 to September 2019 were identified. Pan-TRK immunohistochemistry, and the ALK and ETV6 gene break-apart fluorescence in situ hybridizations (FISH) were performed. NTRK gene rearrangement was detected using sequencing-based methods. Results: There were 3 males and 2 females in this study. The patients were between 3 months and 13 years of age. Histologically, the tumors were infiltrative spindle cell tumors with variable accompanying inflammatory cells. Immunohistochemistry showed positive reactivity for pan-TRK in all tumors, with nuclear staining for NTRK3 fusion, and cytoplasmic staining for NTRK1 fusion. The molecular testing revealed NTRK gene fusions (one each of TPM3-NTRK1, ETV6-NTRK3 and DCTN1-NTRK1, and two cases of LMNA-NTRK1). Two patients were receiving larotrectinib. The others were are well without disease, with follow-up durations of 9 to 29 months. Conclusions: NTRK-rearranged mesenchymal tumors from soft tissue sites and kidney are identified. A novel DCTN1-NTRK1 fusion is described. Pan-TRK immunohistochemistry is useful for diagnosis. NTRK-targeted therapy may be an option for unresectable, recurrent or metastatic cases.

KLF11 promotes gastric cancer invasion and migration by increasing Twist1 expression.

Gastric cancer (GC) is a leading cause of global cancer-related death. The incidence and mortality rates of gastric cancer in China are second and third ranked in all forms of malignant tumors. Krüppel-like factor11 (KLF11) is a member of the KLF family, and previous studies have shown it significantly influences epithelial ovarian, pancreatic and liver cancer proliferation, differentiation and apoptosis. However, the expression and some biological functions of KLF11 in GC are still unclear. We therefore collected and analyzed the mRNA and protein expressions of KLF11 in 59 paired gastric cancer tissues and matched healthy gastric tissue samples. We then investigated the KLF 11 biological functions and potential mechanisms in BGC823 and HGC27 gastric cancer cell lines. Analysis of KLF11 in gastric cancer specimens confirmed up-regulation compared to adjacent healthy gastric tissues, and similar results were evident in the GC cell lines. Ectopic expression of KLF11 was significantly associated with GC cell invasion and migration. KLF11 functions were most effective in Twist1 expression and knockdown, and also in KLF11 up-regulation which was accompanied by corresponding change in Twist1 expression; but these effects were inhibited when KLF11 was silenced by the small interfering RNA (siRNA). The relative Twist1 promoter region activity increased gradually with increasing KLF11 plasma, and KLF11 therefore has a critical role in regulating gastric cancer migration and invasion by increasing Twist1 expression. Finally, the results of this study should improve understanding of the KLF11 and EMT regulating network and KLF11's use as a potential therapeutic target in gastric cancer.

[A reevaluation of diagnostic efficacy of International Society of Thrombosis and Haemostasis and Japanese Association for Acute Medicine criteria for the diagnosis of sepsis disseminated intravascular coagulation].

Objective: To reevaluate the diagnostic efficacy of International Society of Thrombosis and Haemostasis (ISTH) and Japanese Association for Acute Medicine (JAAM) criteria for sepsis disseminated intravascular coagulation (DIC). Methods: A total of 769 patients diagnosed as sepsis were enrolled in our study. Blood samples were collected within the first hour in ICU and the index of coagulation was detected. The correlation between the conventional coagulation index and the acute physiology and chronic health evaluation (APACHE Ⅱ) and sequential organ failure assessment (SOFA) scores was analyzed. The sensitivity and specificity of diagnostic efficacy were analyzed by receiver operating characteristic (ROC) curve. Results: In the 769 cases, 95 cases (12.35%) conformed to the standard of ISTH and 271 cases (35.24%) were in accordance with the standard of JAAM. Prolonged prothrombin time (PT) was seen in 726 cases (94.41%). Activated partial thromboplastin time (APTT) was prolonged in 434 cases (56.44%). Plasma fibrinogen (Fib) was decreased in 94 cases (12.22%) and increased in 365 cases(47.46%). Platelet (PLT) count decreased in 158 cases (20.55%). D-dimer was elevated in 759 cases (98.70%). Fibrin degradation product (FDP) was increased in 724 cases (94.15%). PT, APTT, D-dimer, FDP, PLT were correlated with APACHE Ⅱ(r value were 0.259, 0.348, 0.319, 0.289,-0.275, all P values<0.05) and SOFA score(r values were 0.409, 0.445, 0.407, 0.411,-0.526, respectively, all P values<0.05). The areas under the curve (AUCs) in the ISTH standard from high to low were accordingly PT (0.813), FDP (0.792), PLT (0.746), Fib (0.563). The AUCs from high to low were FDP (0.844), PLT (0.716), and PT (0.660), respectively in the JAAM standard. Under the criteria of ISTH, the diagnostic sensitivities of PT, PLT, Fib and FDP were 92.63%, 67.37%, 9.47%, 98.95%, respectively, and specificities as 53.56%, 86.05%, 99.26% and 33.38%% respectively. As to the JAAM criteria, the diagnostic sensitivities of PT, PLT, and FDP were 74.54%, 52.77%, 91.51% and specificities as 51.61%, 84.94%, 40.76% respectively. Conclusions: According to the ISTH and JAAM diagnostic criteria, the diagnostic efficacy of PT and PLT is relatively high, which is associated with the severity of DIC. D-dimer and FDP have the high sensitivity but the specificity is poor. The diagnostic specificity of Fib is good, yet with low sensitivity and poor overall efficacy.

[High definition MRI rectal lymph node aided diagnostic system based on deep neural network].

Objective: To investigate the clinical significance of high definition (HD) MRI rectal lymph node aided diagnostic system based on deep neural network. Methods: The research selected 301 patients with rectal cancer who underwent pelvic HD MRI and reported pelvic lymph node metastasis from July 2016 to December 2017 in Affiliated Hospital of Qingdao University. According to the chronological order, the first 201 cases were used as learning group. The remaining 100 cases were used as verification group. There were 149 males (74.1%) and 52 females in the study group, with an average age of 58.8 years. There were 76 males (76.0%) and 24 females in the validation group, with an average age of 60.2 years. Firstly, Using deep learning technique, researchers trained the 12 060 HD MRI lymph nodes images data of learning group with convolution neural network to simulate the judgment process of radiologists, and established an artificial intelligence automatic recognition system for metastatic lymph nodes of rectal cancer. Then, 6 030 images of the validation group were clinically validated. Artificial intelligence and radiologists simultaneously diagnosed all cases of HD MRI images and made the diagnosis results of metastatic lymph node. Receiver operating characteristic (ROC) curve and area under curve (AUC) were used to compare the diagnostic level of them. Results: After continuous iteration training of the learning group data, the loss function value of artificial intelligence decreased continuously, and the diagnostic error decreased continuously. Among the 6 030 images of verification group, 912 images were considered to exist metastatic lymph nodes in radiologists' diagnosis and 987 in artificial intelligence diagnosis. There were 772 images having identical diagnostic results of lymph node location and number of metastases with the two methods. Compared with manual diagnosis, the AUC of the intelligent platform was 0.886 2, the diagnostic time of a single case was 10 s, but the average diagnostic time of doctors was 600 s. Conclusion: The HD MRI lymph node automatic recognition system based on deep neural network has high accuracy and high efficiency, and has the clinical significance of auxiliary diagnosis.

Relationship between Expression Changes of CB2R and Wound Age of Brain Contusion in Mice.

ABSTRACT: Objective To investigate the expression of cannabinoid type 2 receptor （CB2R） at different time points after brain contusion and its relationship with wound age of mice. Methods A mouse brain contusion model was established with PCI3000 Precision Cortical Impactor. Expression changes of CB2R around the injured area were detected with immunohistochemical staining, immunofluorescent staining and Western blotting at different time points. Results Immunohistochemical staining results showed that only a few cells in the cerebral cortex of the sham operated group had CB2R positive expression. The ratio of CB2R positive cells gradually increased after injury and reached the peak twice at 12 h and 7 d post-injury, followed by a decrease to the normal level 28 d post-injury. The results of Western blotting were consistent with the immunohistochemical staining results. Immunofluorescent staining demonstrated that the changes of the ratio of CB2R positive cells in neurons, CB2R positive cells in monocytes and CB2R positive cells in astrocytes to the total cell number showed a single peak pattern, which peaked at 12 h, 1 d and 7 d post-injury, respectively. Conclusion The expression of CB2R after brain contusion in neurons, monocytes and astrocytes in mice suggests that it is likely to be involved in the regulation of the biological functions of those cells. The changes in CB2R are time-dependent, which suggests its potential applicability as a biological indicator for wound age estimation of brain contusion in forensic practice.

Three-vessel-trachea view in the diagnosis of fetal cardiac great vessel malformation.

Fetal cardiac great vessel malformation is attracting increasing attention in the prenatal ultrasonic diagnosis of fetal congenital heart disease. To investigate the clinical diagnostic values of three-vessel-trachea view (3VT view) in the ultrasonic diagnosis of this malformation, the present study analyzed the echocardiographic examination results of 77 fetuses with great vessel malformation, retrospectively analyzed the echocardiographic characteristics in the three-vessel-trachea view, and followed up the enrolled cases. The results suggest that great vessel malformation had characteristic manifestations, such as abnormal arrangement order, inner diameter, blood flow direction and branch. Color Doppler flow imaging found V, O, C, U, Ioo and oVo structures. There were 20 cases of blood vessel position abnormality, 38 cases of abnormal blood vessel diameter, and 19 cases of abnormal number of blood vessels. The detection rate of abnormal blood vessel diameter was 95%, which was the highest; the detection rate of abnormal blood vessel position was 97.4%, and that of abnormal number of blood vessels was 84.2%. It is concluded that the 3VT view can indicate fetal cardiac great vessel malformation. The 3VT view is beneficial to timely prenatal diagnosis, relief of body pain and improvement of quality of birth.

[Changes and differences of DNA methylation in human macrophages infected with virulent and avirulent Mycobacterium tuberculosis].

Objective: To profile DNA methylation and compare differentially methylated region (DMR) of macrophages infected with virulent and avirulent strains of Mycobacterium tuberculosis (MTB). Methods: PMA(50 ng/ml) treated THP-1 macrophages were left uninfected or infected with the virulent H37Rv(THP-1/Rv) or avirulent H37Ra(THP-1/Ra). The genomic DNA was then extracted and DNA methylation was profiled via Reduced Representation Bisulfite Sequencing (RRBS). The DNA methylation pattern and DMRs between the tested samples were indentified. DMRs-associated genes were annotated, clustered by Gene Ontology (GO), and the pathways analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG). mRNA expression of several DMR-associated genes were verified by Q-PCR, and Student's t-test was used to analyze the differential expression between either 2 samples. Results: The mostly DNA methylated C in 3 samples was in CG, accounting for 98% of all types of methylated C(CG, CGH, CHH). Besides, the average DNA methylation level in THP-1/Rv, THP-1/Ra and THP-1/NC were 7.72%, 7.38% and 7.58% in the promoter, and 9.90%, 9.57% and 9.80% in the CpG island (CGI), respectively. We identified 58 DMRs between H37Rv and H37Ra infected THP-1 macrophages with an average length of 152 bp. Six DMRs-associated genes were enriched. Among them, AP-1 and DDIT4 were differentially expressed in cells infected with H37Rv and H37Ra. Conclusions: Infection with MTB is not correlated with a large-scale gain or loss in DNA methylation in host cells. AP-1 and DDIT4 might be novel virulence-related genes regulated by DNA methylation of their corresponding promoter regions.

[Modified capsular arthroplasty for young patients with developmental dislocation of the hip].

Developmental dislocation of the hip(DDH) is one of the most serious hip diseases. Patients with unilateral DDH are prone to secondary osteoarthritis, low back pain, gait disturbance and compensatory scoliosis because of the leg length discrepancy. Total hip arthroplasty(THA) is the highly effective treatment for patients with hip pain or dysfunction caused by unilateral DDH, thus decrease the demand for hip-preserving surgeries such as capsular arthroplasty which may postpone or avoid hip replacement. However, the failure rate of THA for young patients is high and the majority of young patients may require one or more revision arthroplasties throughout their lifetime. The basic principle of capsular arthroplasty is that the femoral head wrapped by capsule is placed into a newly reamed socket on the location of true acetabulum. Therefore, hip replacement for patients with previous capsular arthroplasty becomes easier and safer than primary THA. However, the early capsular arthroplasty have been abandoned due to the relatively high rates of femoral head necrosis and hip stiffness. Ganz modified the early procedure with the technique of surgical hip dislocation in 2012, and emphasized the importance of postoperative rehabilitation. The incidence of complication, therefore, decreased greatly due to the preservation of main blood supply of femoral head as well as the proper postoperative management. In order to improve the clinical outcomes of this modified procedure, the selection of indications and surgeons' experience should also be taken into consideration.

[A retrospective clinical study of sixty-three cases with persistent inflammation immunosuppression and catabolism syndrome].

Objective: To study the clinical characteristics and prognosis of patients with persistent inflammation immunosuppression and catabolism syndrome (PICS) in ICU. Methods: A total of 126 patients admitted to ICU (ICU stay of more than 10 days, age≥18 years) between January 2014 to December 2014 were retrospectively studied.Data were collected from electronic medical records including demographics, underlying disease, Acute Physiology and Chronic Health Evaluation Ⅱ (APACHEⅡ) score, Sequential Organ Failure Assessment (SOFA) score, laboratory parameters, ICU acquired infections and clinical outcome. Results: The overall incidence of PICS in ICU patients (ICU stay of more than 10 days) was 50.0%(63/126). There were no significant differences in baseline data such as gender, age, APACHEⅡscore, SOFA score and underlying diseases between the two groups(all P>0.05). Compared with the non-PICS group, there were more patients with gastrointestinal perforation in the PICS group (P=0.042), however, the medical or surgical admission did not differ between the two groups(P>0.05). During the stay in ICU, the PICS group showed a higher risk of developing acquired infections compared with the non-PICS group[PICS 63.5%(40/63) vs non-PICS 23.8%(15/63); P<0.001]. The infections were more caused by Candida in the PICS group than the non-PICS group [PICS 22.4%(11/49) vs non-PICS 2/17; P=0.003]. Moreover, the PICS group experienced longer stay in ICU[PICS(31.6±28.8) days vs non-PICS (20.4±11.3) days; P=0.0046] and higher ICU mortality [PICS 28.6%(18/63) vs non-PICS 6.3%(4/63), P=0.001]. Conclusion: PICS is a common manifestation of patients who stay in ICU more than 10 days, which is associated with high risk of ICU acquired infections, prolonged length of stay and high mortality.

[DNA damage associated genetic variants contribute to lung cancer susceptibility in a Han Chinese population].

OBJECTIVE: To explore the association between DNA damage-related genetic variants and lung cancer susceptibility in a Han Chinese population. METHODS: This case-control study enrolled patients from the Cancer Hospital of Jiangsu Province and Jiangsu Province Hospital from 2003 to 2009. Controls were randomly selected from individuals who visited the same hospital or a community-based health examination program during the same time period. A 5 ml venous blood sample was obtained from each participant and epidemiological information was collected on a standard questionnaire. Illumina Infinium(®) BeadChip was used for genotyping of 35 DNA damage-related single nucleotide variations (SNVs), which were identified in our previous study. Multivariate and binary logistic regressions were used to calculate the OR and 95%CI for lung cancer risk. HaploReg V4.1 and Regulome DB were used to understand functional annotation on important SNV. RESULTS: The distributions of age (61.06±10.15) vs. (61.32±11.07) years; t=-0.72, P=0.473) and sex (χ(2)=1.81, P=0.179) were similar between cases and controls. However, the case group had a higher frequency of smokers (61.08% vs. 48.54%; χ(2)=50.04, P<0.001) and heavy smokers (42.28% vs. 24.07%; χ(2)=122.32, P<0.001). Among the 34 SNVs that passed quality control, two SNVs were significantly associated with lung cancer risk after adjustments for age, sex and cumulative smoking dose: rs9267576 C>A (CA genotype/CC genotype, OR=1.56, 95% CI: 1.01-2.40) and rs3130683 A>G (AG genotype/AA genotype, OR=1.87, 95%CI: 1.13-3.09). After step-wise logistic regression analysis, only the rs3130683 SNV was retained in the model, indicating that the association between rs9267576 and lung cancer may be due to the effect of rs3130683. Functional annotation indicated that rs3130683 was located in the promoter and enhancer regions, and was an expression quantitative trait loci of HLA. The Cancer Genome Atlas indicated that expression of HLA-C, DQB1, DRB1 and DRB5 in lung cancer tissue was significantly lower than in paired normal tumor-adjacent tissue, with down-regulation of the four respective genes in 81.3%, 88.8%, 90.7% and 90.7% of lung cancer tissues (P-values were 6.68×10(-15), 2.21×10(-13), 2.20×10(-16), 2.58×10(-13), respectively). CONCLUSIONS: The SNV rs3130683 (A>G) was associated with the risk of lung cancer in a Han Chinese population. This SNV may affect the risk of lung cancer by regulating HLA expression.

[Novel biomarkers for latent tuberculosis infection by plasma proteomic profiling].

OBJECTIVE: To screen specific biomarkers for latent tuberculosis infection by comparing the plasma proteomic profiling between latent tuberculosis infection and healthy controls. METHODS: The plasma proteins from 15 cases with latent tuberculosis infection and 15 healthy controls were detected by the label-free quantitative proteomic technology. Differential expressed proteins were analyzed by GO, KEGG, and BiNGO analysis. Student's t test was used to analyze the differential expression between 2 groups. RESULTS: Twenty-three candidate proteins were identified, among which 15 proteins were downregulated (<0.5-fold at P<0.05) and 8 proteins were upregulated (>2.0-fold at P<0.05) in the latent tuberculosis infection group. Bioinformatic analysis revealed 3 proteins AAT, C3 and C4A to be the most significant. CONCLUSION: There were differential plasma protein profiles between latent tuberculosis infection and healthy controls. Candidate proteins AAT, C3 and C4A were promising biomarkers for discriminating cases with latent tuberculosis infection from healthy persons.