Aim To study and compare the pharmaco-kinetic parameters of roxithromycin under normoxic and hypoxic rats. Methods A highly effective and rapid ultra-performance liquid chromatography with tandem mass spectrometry ( UPLC-MS/MS) method with posi-tive electrospray ionization source was successfully de-veloped and validated for quantification of roxithromy-cin in rat plasma. Sprague-Dawley rats were randomly divided into the hypoxia and normoxic groups. Each rat obtained a single dose of roxithromycin with 10 mg · kg-1 via intragastric administration. The pharmacoki-netic parameter comparison between normoxic and hy-poxic groups was calculated by SPSS software using in-dependent sample t test method. Results The main pharmacokinetic parameters of roxithromycin between the normoxic and hypoxic rats were:the AUC(0-t) 7 576 and 3 761 μg·h·L-1 , MRT(0-t) 5. 6 and 7. 7 h, T1/2 3. 4 h and 3. 9 h, CL 1. 5 and 3. 0 L · h-1 · kg-2 , tmax3. 1 and 3. 4 h, Cmax 1 116 and 372 μg·L-1 , re-spectively. The levels of Cmax and AUC of roxithromy-cin in hypoxic rats were statistically lower than those in normoxic rats. Conclusion The exposure level of rox-ithromycin in hypoxic rats markedly decreased. Our re-sults may provide an important experimental basis to adjust the dosage for roxithromycin in hypoxic clinical practice.

Objective:To compare the clinical effect of Ilizarov frame and unilateral frame in the treatment of tibial segmental bone defect combined with soft tissue defect by open bone transport.Methods:A retrospective case-control study was conducted to analyze the clinical data of 32 patients with tibial segmental bone defect and soft tissue defect-up who underwent open bone transport technique in Tianjin Hospital from August 2008 to August 2016 and obtained complete followed-up. There were 22 males and 10 females, aged 22-64 years (mean, 36.8 years). Nineteen patients in the Ilizarov group showed the mean bone defect length of 7.9 cm and mean soft tissue defect area of 41.4 cm 2. Thirteen patients in the unilateral group showed the mean bone defect length of 7.8 cm and the mean soft tissue defect area of 39.2 cm 2. The results of fracture healing, wound healing time, radiological consolidation index, external fixation index, Association for the Study and Application of the Method of Ilizarov (ASAMI) bone and lower-limb function score and incidence of complications were compared between the groups. Results:All patients were followed up for 24 to 60 months [(32.6±1.3)months]. Bone healing was achieved in all patients. The wound healing time was (2.7±2.3)days in Ilizarov group and (2.4±1.8)days in unilateral group ( P>0.05). The consolidation index was (43.4±8.7)d/cm in Ilizarov group and (45.8±10.3)d/cm in unilateral group ( P>0.05). The external fixation index was (52.6±8.9)d/cm in Ilizarov group and (58.7±12.9)d/cm in unilateral group ( P<0.05). The results of ASAMI bone score was excellent in 10 patients, good in 6, fair in 2 and poor in 1 in Ilizarov group, with the excellent rate of 84%; and excellent in 6 patients, good in 4, fair in 2, and poor in 1 in unilateral group, with the excellent rate of 77%, with insignificant difference between two groups ( P>0.05). The results of ASAMI lower-limb function score was excellent in 8 patients, good in 7, fair in 3, poor in 1 in Ilizarov group, with the excellent rate of 79%; and excellent in 6, good in 3, fair in 3, poor in 1 in unilateral group, with the excellent rate of 69%, with insignificant difference between the two groups ( P>0.05). Incidence of axial deviation was zero in Ilizarov group versus 23% in unilateral group ( P<0.05). After operation, Ilizarov group had refracture in 1 patient and pin site infection in 5, and unilateral group had refracture in 1 patient and pin site infection in 1, showing no significant difference between the two groups ( P>0.05). Conclusions:Tibial segmental bone defect with soft tissue defect can be effectively treated by open bone transport with Ilizarov and unilateral frame. Ilizarov frame has better biomechanical properties and is more convenient for correction of poor postoperative axial alignment.

Objective:To investigate the accuracy and postoperative efficacy of fluoroscopy and CT in measuring the mounting parameters of Taylor Spatial Frame.Methods:Data of patients with peripheral knee deformities who were treated by Taylor Spatial Frame from June 2006 to December 2017 were retrospectively analyzed. According to different measurement methods of mounting parameters, they were divided into fluoroscopy group (mounting parameters were obtained by intraoperative fluoroscopy) and CT group (mounting parameters were obtained by postoperative CT). There were 33 patients (35 segments) in the fluoroscopy group, 23 males (23 segments) and 10 females (12 segments), with an average age of 36.4±11.6 years old. In CT group, there were 30 patients, 19 males and 11 females, with an average age of 36.9±13.8 years. There were 22 cases (24 segments) of high tibial osteotomy, 5 cases (5 segments) of distal femur osteotomy, and 6 cases (6 segments) of both distal femur and high tibial osteotomy. Operation time, external fixation time, the number of electronic prescription and deformity correction time, mechanical axis deviation (MAD), medial proximal tibia angle (MPTA), mechanical lateral distal femoral angle (mLDFA), range of motion (ROM) andhospital for special surgery (HSS) knee functional scores were compared between the two groups.Results:All the 63 patients were followed up for 21.9 months (range, 12-60 months). In fluoroscopy group, operating time was 100.9±9.1 min, electronic prescription number 1.4±0.6, and deformity correction time was 19.4±3.6 days. In CT group, operating time was 79.2±10.8 min, electronic prescription number 1.2±0.4, and deformity correction time was 16.0±4.4 days. The difference of the above indexes between the two groups was statistically significant ( t=8.803, 2.042, 3.440, all P < 0.05). In the fluoroscopy group, the external fixation time was 4.8±0.9 months; MAD was 4.3±2.1 mm; MPTA was 88.5°±1.9°; mLDFA was knee 89.2°; ROM was 122.4°±3.9° and HSS score was 90.0±3.6. In CT group, the external fixation time was 4.6±0.9 months; MAD was 4.0±1.9 mm; MPTA was 87.8°±1.7°; mLDFA was knee 88.6°; ROM was 122.7°±3.4° and HSS score was 91.1±2.9. There was no statistically significant difference in the above indexes between the two groups (all P >0.05). In the fluoroscopy group, 22 segmental deformities were corrected by one electronic prescription, and 13 segmental deformities were corrected by two or more electronic prescriptions. In CT group, 25 segmental deformities were corrected by one electronic prescription, and 5 segmental deformities were corrected by two electronic prescriptions. There was no incision infection and no neurovascular injury in the two groups. Conclusion:Both fluoroscopy and CT scan can obtain the mounting parameters of the Taylor Spatial Frame, and the results of correction of the peripheral deformities of the knee joint are satisfactory. However, CT measurement of the mounting parameters is more accurate which could achieve shorter operation time, and less times of electronic prescriptions.

OBJECTIVE To compare the metabolomic characteristics of stage T1 papillary thyroid carcinoma(PTC)and nodular goiter(NG),and the relationship between metabolites and lymph node metastasis of PTC.METHODS Serum samples were collected from 60 patients with stage T1 PTC and 30 patients with NG who underwent thyroidectomy at the Department of Otolaryngology Head and Neck Surgery,Civil Aviation General Hospital between September 2021 and April 2022.The PTC group was divided into the N+ group with lymph node metastasis and the N-group without lymph node metastasis according to the presence or absence of lymph node metastasis.The serum metabolites of the N+ and N-groups and the PTC and NG groups were compared and analyzed using an ultra-performance liquid chromatography-mass spectrometry(UPLC-Q-Exactive-MS)coupled platform,and principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA),and orthogonal partial least squares discriminant analysis(OPLS-DA)was performed using SIMCA-P 14.1 software.OPLS-DA modeling,combined with FDR-corrected Mann-Whitney-Wilcoxon test results and metabolite difference multiples in the two groups undergoing comparison,etc.to screen for potential small molecule metabolic markers,and to establish a joint diagnostic model by binary logistic regression analysis.RESULTS There were no significant differential metabolites between the N+ group with lymph node metastasis and the N-group without lymph node metastasis.Seven differential metabolites were found between PCA patients and NG patients,and the five relevant metabolic pathways were the pentose phosphate pathway,pentose and glucuronide interconversion,glycolysis/gluconeogenesis,fructose,and mannose metabolism,and fatty acid biosynthesis.The differential metabolite with an area under the ROC curve>0.9 was D-glyceraldehyde 3-phosphate,and another N-undecanoylglycine,uronic acid,and the area under the ROC curve for three metabolites,N-undecanoylglycine,uric acid,and triiodothyronine glucuronide,was>0.8.CONCLUSION PTC patients differed from NG patients mainly in glucose metabolism and lipid metabolism,and D-glyceraldehyde 3-phosphate could be distinguished from NG patients with the aid of N-undecanoylglycine,uric acid,and triiodothyronine glucuronide,combined with imaging findings.Also,no significant differences in serum metabolites were found in the N+ group compared with the N-group,and the presence or absence of lymph node metastases did not affect serum metabolites in patients with stage T1 PTC.

Objective To test and evaluate the JAK2 gene V617F mutation in patients with myeloproliferative tumors based on i-densy IS-5320 platform according to ISO15189 accreditation requirements.Methods Instrument performance verification.Selected from December 2014 to February 2017, 20 cases of JAK2 V617F mutation positive peripheral blood samples from Huashan Hospital of the Shanghai FuDan University Medical College and 20 cases of peripheral blood samples with negative JAK2V617F mutation.The Realtime PCR with TaqMan MGB probe was selected as the control method to verify and evaluate the accuracy of testing JAK 2 V617F mutation on i-densy IS-5320 platform.Whole blood samples were used to evaluate the reproducibility , cross-contamination and anti-interference ability of this platform.The ability of mutation load was verified by detecting mixtures of human erythnoleukemia cells and colorectal cancer cell HCT116 with 12 different proportions.Results I-densy IS-5320 platform and TaqMan MGB probe real-time fluorescence quantitative PCR show the same result .The within-run reproducibility and between-run reproducibility are both 100%.There is no observed contamination .High bilirubin and high triglyceride blood samples have no obvious interference on mutation detection .The mutation ratio with a load as low as 0.25%could be tested stably by i-densy IS-5320 platform.The detecting peak of melting curve can reflect the ratio of JAK2 V617F mutation to some extent.Conclusions I-densy IS-5320 can detect the mutation of JAK2 V617F gene in the whole blood directly.It has high sensitivity, accuracy and stability, and is easy to operate, and also can reflect the mutation load of JAK2 V617F, which could meet the clinical requirements for the detection of mutations.

Objective By a sequencing panel consisting of 50 targeted genes, aiming at depicting the molecular landscape of ET, PV, and PMF, which are three major subtypes of MPN, to provide valuable information in the diagnosis and prognosis of MPN.Methods A retrospective study was conducted of 53 patients from Huashan hospital and Changhai hospital. All patients were diagnosed in accordance with the 2016 WHO diagnostic criteria for MPN, including 31 cases of ET(11 males, 20 females, median age 55 years), 17 cases of PV(12 males, 5 females, median age 65 years), and 5 cases of PMF(4 males, 1 females, median age 67 years), and underwent next-generation of DNA sequencing of their bone marrow or blood samples. The genetic analyses were performed on bone marrow or peripheral blood. Referring to COSMIC, dbSNP, Clinvar and other public databases, we analyzed the sequencing data, and elucidated the mutation profile of MPN patients, combining with their clinic information. Results In addition to the typical JAK2, CALR, and MPL mutations, pathogenic mutations in other 11 genes were detected, as well as 4 SNPs that confer individual susceptibility to MPNs (rs4858647, rs9376092, rs58270997, rs621940). The average rate of mutated genes was 2.3 genes per patient. In all patients (53 cases), the mutated genes detected were TET2, EZH2, ASXL1, MIR662, SF3B1, BARD1, DNMT3A, KIT, RUNX1, TP53, NRAS according to their mutational frequency. Conclusions Applying next-generation sequencing technology, multi-gene sequencing of a bunch of typical BCR-ABL-negative MPN patients can be performed at one time within 2 working days, and pathogenic mutations other than JAK2, CALR, MPL can be found, which has a bright prospection in clinic.

OBJECTIVE: To explore the clinical characteristics and genetic etiology of a child with multiple pterygium syndrome (MPS). METHODS: A child with MPS who was treated at the Orthopedics Department of Guangzhou Women and Children's Medical Center Affiliated to Guangzhou Medical University on August 19, 2020 was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and her parents were also collected. Whole exome sequencing (WES) was carried out for the child. Candidate variant was validated by Sanger sequencing of her parents and bioinformatic analysis. RESULTS: The child, an 11-year-old female, had a complain of "scoliosis found 8 years before and aggravated with unequal shoulder height for 1 year". WES results revealed that she has carried a homozygous c.55+1G>C splice variant of the CHRNG gene, for which both of her parents were heterozygous carriers. By bioinformatic analysis, the c.55+1G>C variant has not been recorded by the CNKI, Wanfang data knowledge service platform and HGMG databases. Analysis with Multain online software suggested that the amino acid encoded by this site is highly conserved among various species. As predicted with the CRYP-SKIP online software, the probability of activation and skipping of the potential splice site in exon 1 caused by this variant is 0.30 and 0.70, respectively. The child was diagnosed with MPS. CONCLUSION The CHRNG gene c.55+1G>C variant probably underlay the MPS in this patient.
Humans ; Child ; Female ; Abnormalities, Multiple/genetics* ; Malignant Hyperthermia/genetics* ; Skin Abnormalities/genetics* ; Heterozygote ; Mutation ; Receptors, Nicotinic/genetics*

OBJECTIVE： To study the anti-gout effect of aqueous extract from the stems and leaves of Erythropalum scandens （ASLE）. METHODS： The mice were randomly divided into normal group， model group， allopurinol group （positive control， 5 mg/kg）， ASLE low-dose， medium-dose and high-dose groups （1 300， 2 600， 5 200 mg/kg， by raw material； similarity hereinafter）， with 10 mice in each group. Except for normal group， other groups were given potassium oxonate intragastrically to induce hyperuricemia model. One hour after modeling， normal group and model group were given constant volume of normal saline intragastrically； administration group was given relevant medicine intragastrically， once a day， for consecutive 7 d. One hour after last administration， the levels of serum uric acid （SUA） and serum creatinine （Scr） were detected by colorimetry assay. Another mice were randomly divided into normal group， model group， indomethacin group （positive control， 7.5 mg/kg）， ASLE low-dose， medium-dose and high-dose groups， with 10 mice in each group. Normal group and model group were given constant volume of normal saline intragastrically； administration group was given relevant medicine intragastrically， once a day， for consecutive 7 d. After last administration， except for normal group， the mice were given sodium microcrystalline urate via toes to induce gouty arthritis model. Before and 1， 2， 4， 6， 8 h after modeling， the circumference of the same part of the inflamed limbs and toes of mice in each group was measured by wire binding method， and the degree of toe swelling was calculated. The number of white blood cell （WBC）， neutrophil （NEU） and lymphocyte （LYM） were detected by animal hematology analyzer. The levels of SUA and Scr were measured by colorimetry assay. The content of NO in toe tissue was determined by Griess method. RESULTS： The experimental results of hyperuricemia model showed that the levels of SUA and Scr in mice were significantly higher in model group than those in normal group （P＜0.01）. Compared with model group， above indexes of mice were decreased significantly in administration group （P＜0.05 or P＜0.01）. The experimental results of gouty arthritis model showed that the level of SUA， the degree of toe swelling （2-8 h）， the number of WBC， NEU and LYM， NO content in model group were increased significantly， compared with normal group （P＜0.05 or P＜0.01）. Compared with model group， the levels of SUA and Scr （ASLE groups）， the degree of toe swelling [indomethacin group， ASLE high-dose group （2-8 h）， ASLE low-dose group （2， 6 h）， ASLE medium-dose group （6 h）]， the number of WBC and NEU （administration groups）， the number of LYM （indomethacin group） and NO content （administration groups except for ASLE low-dose group） were decreased significantly in administration groups （P＜0.05 or P＜0.01）. CONCLUSIONS： The anti-gout effect of ASLE may be associated with promoting uric acid metabolism， anti-inflammatory and improving renal function.

Immune escape is one of the ten hallmarks of tumors， which plays an important role in the occurrence and development of tumors. Immune escape refers to a process where tumor cells remodel and edit the immune system through the model of immune clearance， immune balance， and immune escape to "transform" the immune cells into immunosuppressive cells in the tumor microenvironment， so as to support immune escape. The five-stage evolution is the summary of tumor pathogenesis by professor LI Jie. He believes that the gradual development of tumors follows the core pathogenesis of "deficiency-cold-toxin-obstruction-collapse"， in which "depression" runs through the whole process， and cancer toxin is the key. Based on immune editing， this paper combined phenotypic characteristics of tumor cells with the core pathogenesis of the five-stage evolution of professor LI to reveal the biological basis of malignant tumor five-stage evolution. The results indicate that the prominent change from deficiency to cold is the reduction of immune surveillance and the prominent change from toxin to obstruction is immune escape. The final stage of collapse is the outcome of immune failure. Depression is the booster of tumor immune editing. Therefore， the method of reinforcing the healthy Qi and removing toxins was proposed to regulate the immune editing and cut off the five-stage evolution of tumors. Supplementing Qi and warming Yang can reinforce the healthy Qi and restore immune surveillance. Removing toxins and dredging can reverse toxins and immune escape. The harmonizing method can maintain the dynamic balance of immune cells/immunosuppressive cells. Resolving depression can truncate tumor immune editing. Those methods can provide a certain reference for the treatment based on microscopic syndrome differentiation in traditional Chinese medicine （TCM）. In future studies， it is necessary to further explore the specific mechanism of the regulation of immune editing with the methods of supplementing Qi and warming Yang， removing toxins and dredging， their combination， and resolving depression， so as to find out specific Chinese medicines and targets and provide more sufficient evidence for the regulation of tumor immune editing by TCM.

Objective: To investigate the diagnosis, treatment and prognosis of nonodontogenic periapical lesions and to provide a reference for clinical diagnosis and treatment. Methods: A case of a patient with right upper molar pulp with apical penetration and local occlusion admitted to the West China Stomatological Hospital of Sichuan University was retrospectively analyzed, and the curative effect of microapical surgery and pith preservation was also analyzed. Results : The imaging features of tooth 16 showed periradicular radiolucency combined with local radiopaque lesions around the distal buccal apical area. Endodontic microsurgery was performed under local anesthesia. Soft tissue coverage was observed in the distal buccal apical area during the surgery, and no radiopaque tissue was detected. The distal buccal root apex was cut by 3 mm, and mineral trioxide aggregate was used for root-end backfilling. The postoperative pathological results revealed fibrous connective tissue. One-week recall X-ray examination showed tight root-end backfilling and no periradicular radiolucency; an electrical test of pulp vitality showed positive results. The four-year follow-up showed that there was no discoloration in tooth 16 and no significant difference in thermal and electrical tests of pulp vitality compared with control teeth. Combining the clinical manifestations, imaging features, surgical exploration results and pathological reports, the case was most likely to be cemental hypoplasia. Through the literature review, the treatment and healthy pulp preservation of such cases by endodontic microsurgery under the premise of preserving teeth has not been reported. Conclusion For maxillary posterior teeth with periapical lesions but healthy pulp, accurate estimation of pulp status, endodontic microsurgical exploration and application of bioactive materials can achieve vital pulp preservation while removing the lesions.