Objective To screen the specific cytokines of tuberculous pleural effusion(plTB)by using liquid array technique to establish a diagnostic model and discuss its application value.Methods A total of 86 patients with plTB(plTB group)were included,including 41 patients in the confirmed plTB group and 45 patients in the clinically diagnosed plTB group.There were 42 other patients with pleural effusion in the control group.Seventeen cytokines in pleural effusion were analyzed by liquid array technology.Interleukin(IL)-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-9,IL-10,gamma-interferon-induced protein 10(IP-10),IL-15,IL-17F,IL-27,tumor necrosis factor(TNF)-α,monocyte chemotactic protein-1(MCP-1),the expression levels of macrophage inflammatory protein-3a(MIP-3α),macrophage colony-stimulating factor(M-CSF)and β-interferon(IFN-β)were detected.Difference factors between the confirmed plTB group and the control group were screened,and the receiver operating characteristic(ROC)curve was drawn in the confirmed plTB patients.IP-10,IL-27 and MCP-1 with AUC>0.850 and specificity>80%were combined to diagnose plTB,and were compared with adenylate deaminase(ADA)and T-SPOT.TB in pleural effusion to evaluate the diagnostic efficacy.Results The levels of IL-2,IP-10,IL-27,TNF-α and MCP-1 were higher in the confirmed plTB group than those in the control group(P＜0.05).The sensitivity and specificity of IP-10,IL-27 and MCP-1 in the diagnosis of plTB were 87.8%and 81.0%.The sensitivity of three-factor combined diagnosis in 45 patients with plTB was still as high as 86.7%,and there was no significant difference in sensitivity compared with that in the diagnosed plTB group(P＞0.05).In the plTB group,the sensitivity of IP-10,IL-27 and MCP-1 combined detection was 87.2%,which was higher than that of T-SPOT.TB(81.4%)and ADA(54.7%).Conclusion The application of liquid array technology to the joint detection of pleural effusion IP-10,IL-27 and MCP-1 can provide help for the diagnosis of plTB.

OBJECTIVE: To explore the genetic etiology of a Chinese pedigree affected with pseudohypoparathyroidism. METHODS: Peripheral blood samples of the proband and his parents were collected and subjected to trio-whole exome sequencing (trio-WES). Candidate variants were verified among the pedigree and 50 randomly selected healthy individuals through analysis of restriction fragment length polymorphism. Short tandem repeat (STR) linkage analysis was used to verify the parental origin of the pathogenic variants. RESULTS: Trio-WES and Sanger sequencing showed that the proband and his mother had both harbored a c.121C>G (p.His41Asp) variant of the GNAS gene, which was not found in other family members and the 50 healthy controls. The variant was not found in international databases. Based on guidelines from the American College of Medical Genetics and Genomics, the variant was predicted to be likely pathogenic. CONCLUSION The novel c.121C>G variant of the GNAS gene probably underlay the disease in this pedigree. Above finding has enriched the spectrum of GNAS gene variants.
Female ; Humans ; Pedigree ; East Asian People ; Mothers ; Exome Sequencing ; Pseudohypoparathyroidism/genetics* ; Mutation ; China ; Chromogranins/genetics* ; GTP-Binding Protein alpha Subunits, Gs/genetics*

Objective To investigate the effect of T cell immunoreceptor with Ig and ITIM domains (TIGIT) on the function of CD8⁺ T cells in the lungs of Plasmodium infected mice. Methods The lungs of the mice infected with Plasmodium yoelii were isolated, weighed and photographed after 12 days' infection. After dissolution, lung lymphocytes were isolated, counted and stained, and then the contents of CD8⁺ and TIGIT⁺CD8⁺ T cells were detected by flow cytometry. The expressions of L selectin (CD62L), CD69, programmed death 1 (PD-1), CD25, and C-X3-C motif chemokine receptor 1 (CX3CR1) on TIGIT⁺CD8⁺ T cells were detected by flow cytometry. After stimulation with phorbol 12-myristate 13-acetate (PMA) and ionomycin, the ability of TIGIT⁺CD8⁺T cells to secrete interferon γ(IFN-γ), interleukin 21 (IL-21), IL-4, IL-17, and IL-10 was detected. Results The body mass of mice with Plasmodium infection was reduced. The lungs became darker, and the ratio of the lung mass to body mass was significantly increased. Compared with the normal mice, the percentages and absolute quantity of CD8⁺ and TIGIT⁺CD8⁺ T cells in the lungs of the infected mice were significantly increased. The percentage of TIGIT⁺CD8⁺ T cells expressing CD62L in the infected group was significantly lower, while the percentage of the CD69, PD-1, and CX3CR1 cells were significantly higher than that of TIGIT⁺CD8⁺ T cells from the normal mice. The percentages of TIGIT⁺CD8⁺ T cells secreting IL-21, IL-4, IL-17 and IL-10 cells in the infected group were significantly lower. Conclusion The lung lesions from mice with Plasmodium infection are obvious, the numbers of TIGIT⁺CD8⁺ T cells increase, and these cells express a variety of activation-related molecules, but the ability to secrete cytokines is reduced.
Animals ; Mice ; CD8-Positive T-Lymphocytes ; Cytokines/metabolism* ; Interferon-gamma/metabolism* ; Interleukin-10/metabolism* ; Interleukin-17/metabolism* ; Interleukin-4/metabolism* ; Lung/metabolism* ; Malaria/metabolism* ; Plasmodium yoelii/metabolism* ; Programmed Cell Death 1 Receptor/metabolism*

Objective:To explore the therapeutic effect and mechanism of Dachengqi decoction on patients with mild acute pancreatitis (MAP).Methods:A parallel randomized controlled trial was conducted. Sixty-eight patients with acute pancreatitis (AP) admitted to Shanghai Traditional Chinese Medicine (TCM)-Integrated Hospital from March 2018 to February 2021 were enrolled. Referring to the condition on admission of the patients and whether they agreed to receive the Dachengqi decoction or not, they were divided into conventional treatment group and Dachengqi decoction group according to the principle of 1∶1 equal randomness. Meanwhile, 20 healthy volunteers were recruited as controls. Both groups of patients were treated with octreotide, fasting, gastrointestinal decompression, antipyretic and analgesic, anti-inflammatory, inhibition of gastric acid and pancreatic juice secretion, maintenance of electrolyte balance and other western conventional medicine. The patients in the Dachengqi decoction group received Dachengqi decoction orally on the basis of routine treatment, 100 mL each time, twice a day, for seven consecutive days. The inflammation parameters [white blood cell count (WBC), C-reactive protein (CRP), procalcitonin (PCT), interleukin-6 (IL-6)] before and after treatment and the recovery time of gastrointestinal function (first exhaust time, time to recover bowel sounds, first defecation time) of patients were recorded. 16S rRNA gene sequencing of stool samples was recorded, and normalized data were obtained after quality control and other related processing. The data were subjected to diversity analysis (Alpha diversity and Beta diversity) and linear discriminant analysis effect size analysis (LEfSe analysis) to observe changes in the gut microbiota of MAP patients. Spearman rank correlation coefficient was used to analyze the correlation between inflammatory indexes and microorganisms at the intestinal genus level. Blood, urine, stool samples, renal function, and electrocardiogram (ECG) during treatment of MAP patients were detected to assess the safety of the treatment.Results:Of the 68 patients with AP, 16 were excluded from moderate-severe AP, 4 were not collected or voluntarily abandoned treatment. Finally, 48 patients with MAP were enrolled, 24 in the conventional treatment group and 24 in the Dachengqi decoction group. The inflammation parameters levels at 7 days of treatment in both groups were significantly lower than those before treatment. CRP, PCT and IL-6 levels in the Dachengqi decoction group were significantly lower than those in the conventional treatment group [CRP (mg/L): 8.50 (3.50, 13.00) vs. 16.00 (9.25, 29.75), PCT (μg/L): 0.06 (0.03, 0.08) vs. 0.09 (0.05, 0.11), IL-6 (ng/L): 6.36 (3.96, 10.79) vs. 13.24 (6.69, 18.87), all P < 0.05]. The first exhaust time, time to recover bowel sounds and first defecation time in the Dachengqi decoction group were significantly shorter than those in the conventional treatment group [first exhaust time (days): 1.62±0.65 vs. 2.80±0.65, time to recover bowel sounds (days): 1.13±0.58 vs. 2.31±0.76, first defecation time (days): 3.12±0.75 vs. 4.39±0.76, all P < 0.05]. The analysis of intestinal microflora diversity showed that both the diversity and abundance of microbial communities were the highest in the healthy control group and the lowest in the conventional treatment group. In addition, the coincidence degree of microbial communities in healthy controls and MAP patients was small, while the coincidence degree of MAP patients among different treatment methods was relatively large. LEfSe analysis showed that Dachengqi decoction reduced the relative abundance of Escherichia coli-Shigella and Clostridium erysipelae, and increased the relative abundance of three beneficial bacteria, namely Lactobacillus, Rombutzia and Brutella. In the intestines of MAP patients, Lactobacillus mucilaginus and Lactobacillus conjunctus were significantly enriched. Correlation analysis showed that positive correlations between Escherichia coli- Shigella and the four inflammatory indicators including WBC, CRP, PCT, IL-6 were statistically significant ( r value was 0.31, 0.41, 0.57, 0.43, respectively, all P < 0.05). There was no significant correlation between other bacteria and inflammatory indicators. During the treatment, there was no obvious abnormality in blood, urine and feces, renal function and ECG of MAP patients. Conclusions:Dachengqi decoction could reduce inflammatory responses and promote recovery of intestinal microecological balance and gastrointestinal function in patients with MAP by regulating the composition of intestinal flora. No significant adverse effects were observed during the treatment period.

OBJECTIVE To study the components of ethyl acetate fraction in Qubai tablet ，and its pharmacodynamics on de melanocyte model ，and explore the material basis for anti-vitiligo effect of Qubai tablet. METHODS The ethyl acetate fraction of Qubai tablets was obtained by extraction ，and its components were analyzed by ultra performance liquid chromatography-mass spectrometry（UPLC-MS）. Model control group ，vehicle control group and 8-methoxypsoralen（8-MOP）administration groups （10，50，100，150，200 μmol/L），ethyl acetate fraction administration groups of Qubai tablet （10，50，100，150，200 μg/mL） were set up in the experiment. By establishing the de melanocyte model ，the effects of ethyl acetate fraction of Qubai tablet on de melanocyte were studied from four aspects ：cell number ，cell viability ，melanin formation and tyrosinase activity. RESULTS UPLC-MS component analysis preliminarily determined the structure of 64 compounds in the ethyl acetate fraction of Qubai tablet ， of which 14 compounds were detected in positive and negative ion mode ；psoralen compounds accounted for the largest proportion ， and the content of psoralen chromone chalcone was the highest in positive and negative ion mode. The results of pharmacodynamic study showed that the ethyl acetate fraction of Qubai tablet could increase the number of de melanocytes ，and significantly improve the cell proliferation rate ，the rate of promoting melanin formation and the rate of promoting tyrosinase activity in the process of melanin formation （P＜0.01）. CONCLUSIONS Psoralen compounds may be the material basis for the anti-vitiligo effect of ethyl acetate fraction of Qubai tablet ；good anti-vitiligo effect of ethyl acetate fraction of Qubai tablet may be related to the promotion of tyrosinase activity.

OBJECTIVE To stud y the chemical cons tituents of n-butanol part of Qubai tablet and its pharmacodynamic effect on the model of de melanocyte. METHODS The n-butanol part of Qubai tablet was prepared. The chemical constituents were analyzed by ultra-high performance liquid chromatography-mass spectrometry （UPLC-MS）. Taking mice B 16 melanoma cells as the research object ，the de melanocyte model was established and divided into model group ，positive control different concentration groups（8-methoxypsoralen 10，50，100，150，200 μmol/L），solvent group （diluted with DMSO ）and Qubai tablet n-butanol part different concentration groups （10，50，100，150，200 μmol/L）. The number of cells were observed by inverted microscope ，and the cell proliferation rate ，the rate of melanin production and promotion rate of tyrosinase activity were also detected. RESULTS In the positive and negative ion mode ，53 compounds in the n-butanol part of Qubai tablet were preliminarily determined （29 in the positive ion mode ，33 in the negative ion mode ，overlapping 9），of which coumarins accounted for the largest proportion ， followed by flavonoids. The n-butanol part of Qubai tablet could significantly increase the number of cells ，which was positively correlated with the action time and administration concentration. It could significantly increase the proliferation rate of cells ，the rate of melanin production and promotion rate of tyrosinase activity （P＜0.01）. CONCLUSIONS Coumarins and flavonoids may be the material basis for the anti-vitiligo effect of n-butanol part from Qubai tablet ；anti-vitiligo effect of n-butanol part of Qubai tablet may be realized by promoting tyrosinase activity.

Objective To investigate the effects from head angle and locking times of locking screws on mechanical properties of the screw-plate system, so as to provide a theoretical basis for doctors to select appropriate locking screws and master the locking times in clinical surgical procedure. Methods Locking screws with different head angles were selected to match with locking compression plates in the case of different locking times, and cantilever bending method was used to compare the differences of mechanical properties in each group. Results There were significant differences in failure load and bending stiffness between locking compression plate and locking screw with different head angles (P<0-05). The mechanical properties of the screw-plate system with the best locking degree were optimum. There was no significant difference between 3 time-locking and 1 time-locking (P>0-05). The failure modes of locking screw with different head angles and different locking times were different, and the failure mode of locking screw had a positive correlation with its head angle and locking times. Conclusions The differences in mechanical properties between locking screw with different head angles and locking compression plate cannot be neglected. It is suggested that doctors should choose locking screw and locking compression plate with the best locking degree in clinical procedure, and choose the right torque wrench to lock in multiple times if necessary, thus to prevent screw loosening from affecting recovery of the patients.

Objective:To investigate the correlation between antiplatelet agents and the risk of ruptured intracranial aneurysm.Methods:Patients with intracranial aneurysm admitted to the Department of Neurology, East Hospital Area of Qingdao Municipal Hospital from June to December 2018 were selected retrospectively. The baseline data of patients and the characteristics of intracranial aneurysms were collected. The independent correlation between antiplatelet agents and the risk of ruptured intracranial aneurysm was identified by the univariable analysis and multivariate logistic regression analysis. Results:A total of 90 patients with intracranial aneurysm were included in the study. There were 31 males (34.44%) and 59 females (65.56%). The median diameter of the aneurysm was 4 mm. Forty-six patients taking antiplatelet agents before being diagnosed with intracranial aneurysm, of which 36 taking aspirin, 3 taking clopidogrel, and 7 taking aspirin+ clopidogrel. There were 31 patients (34.44%) with ruptured aneurysm and 59 (65.56%) with unruptured aneurysm. There were statistical differences in the proportion of patients with age <60 years ( P<0.05), diabetes ( P<0.1), ischemic heart disease ( P<0.05), history of previous stroke or transient ischemic attack ( P<0.01), internal carotid artery aneurysm ( P<0.01), anterior communicating artery aneurysm ( P<0.05), posterior communicating artery aneurysm ( P<0.01) and taking antiplatelet agents before diagnosis ( P<0.1) between the ruptured group and the unruptured group. Multivariate logistic regression analysis showed that age <60 years (odds ratio[ OR] 4.116, 95% confidence interval [ CI] 1.337-12.673; P=0.014), anterior communicating artery aneurysm ( OR 5.015, 95% CI 1.155-22.559; P=0.032) and posterior communicating artery aneurysm ( OR 68.796, 95% CI 6.762-699.951; P<0.001) were the independent risk factors for ruptured intracranial aneurysm, and taking antiplatelet agents was an independent protective factor for ruptured intracranial aneurysm ( OR 0.320, 95% CI 0.104-0.992; P=0.048). Conclusions:Taking antiplatelet agents, especially aspirin, does not increase the risk of ruptured intracranial aneurysm, but may be a protective factor of ruptured intracranial aneurysm. Unruptured aneurysms are not contraindications for antiplatelet therapy in patients with clear indications.

Objective:To explore the feasibility of U6 and Cel-miR-39 as reference genes for quantitative detection of microRNA (miRNA) in cerebrospinal fluid (CSF) of tuberculous meningitis (TBM), and validate the difference of miRNAs between tuberculous and viral meningitis (VM).Methods:The remaining CSF specimens after routine examination were collected in Beijing Chest Hospital of Capital Medical University. A total of 36 TBM and 34 VM patients were enrolled based on the information in the medical records. Total RNA were extracted from the CSF samples, and Taqman based real-time quantitative PCR (RT-CR) analysis were performed to determine the concentration of the miRNAs in CSF. GeNorm, NormFinder and Bestkeeper software were used for stability analysis of the two reference genes. 2 -ΔCt method was used to determine the relative gene expression. Accordance of repeated tests was analyzed by Pearson correlation test. Continuous variables were compared by the t-test. Results:Among the 70 samples, the average cycle threshold (Ct) value of U6 was 30.40±3.30, while the average Ct value of Cel-miR-39 was 21.49±0.70. The expression level of Cel-miR-39 was higher than that of U6. Correlation analysis showed good accordance of the repeated tests among the reference genes and target genes analysis in the randomly selected 10 samples ( r>0.931, P<0.001). Based on the analyses results of the three software, including GeNorm, NormFinder and Bestkeeper, Cel-miR-39 presented better stability in RT-PCR analysis and was more suitable as a reference gene for miRNA quantitative determination in CSF sample of TBM patients. The relative expression levels of the three target miRNAs were calculated using Cel-miR-39 as the reference gene, and miR-126-3p (1.13±0.41 vs 3.34±0.82, t=2.452, P=0.016), miR-130a-3p (0.56±0.10 vs 2.59±0.70, t=2.960, P=0.004) and miR-151a-3p (0.64±0.25 vs 2.11±0.33, t=3.536, P<0.001) were showed significant lower expression levels in CSF in TBM group than that in VM group. Conclusions:Cel-miR-39 can be used as a reference gene for quantitative detection of miRNAs in CSF of TBM patients. Significant differences were detected in expression level of miR-126-3p, miR-130a-3p and miR-151a-3p between TBM and VM group.

Objective:To observe and compare the changes of pulmonary function in patients with pulmonary tuberculosis regular treatment for 3 months.Methods:From April 2018 to June 2019, 500 tuberculosis patients who received regular anti tuberculosis treatment in our hospital were selected.The pulmonary function of patients with pulmonary tuberculosis was measured before treatment and at the end of three months; the results of pulmonary ventilation function, lung volume, diffusing capacity, and the value of forced vital capacity (FVC), maximum expiratory volume in 1 second (FEV 1), maximum expiratory volume in 1 second/forced vital capacity (FEV 1/FVC), total lung volume (TLC), residual volume (RV), carbon monoxide diffusing capacity (D LCO) were compared. Results:252 patients with pulmonary tuberculosis were included. Before treatment and at the end of three months, the abnormal pulmonary function results were 204 cases (80.95%) and 193 cases (76.59%), respectively, and the difference was not statistically significant ( P>0.05). Among them, abnormal pulmonary ventilation function is the most common, especially with obstructive, followed by abnormal diffusing capacity. At the end of three months, the proportions of patients with normal pulmonary ventilation function and normal lung volume were higher than that before treatment ( P<0.05), but there was no significant difference in the proportion of normal diffusing capacity before and after treatment ( P>0.05). The values of FVC, FEV 1, TLC and D LCO at the end of three months were higher than those before treatment, and the difference was statistically significant ( t=-6.414, -6.754, -3.863, -3.311, all P<0.01). Conclusions:Most patients with pulmonary tuberculosis have abnormal pulmonary function. At the end of the three months treatment, the normal rates of the pulmonary ventilation function and lung volume as well as the values of FVC, FEV 1, TLC and D LCO in patients with pulmonary tuberculosis were significantly improved compared with those before treatment.