LASS2/TMSG1 overexpression inhibits proliferation and promotes apoptosis of human lung cancer A549 cells possibly by upregulating ceramide and p38 MAPK to activate a signaling cascade / 南方医科大学学报

OBJECTIVE@#To investigate the effects of LASS2/TMSG1 gene overexpression on proliferation and apoptosis of human lung cancer A549 cells and explore the possible mechanism.@*METHODS@#We examined LASS2/TMSG1 expression level in a previously constructed A549 cell line overexpressing LASS2/TMSG1 using Western blotting. The proliferation and apoptosis of the cells were detected using colony-forming assay, CCK-8 assay, Hoechst/PI double staining and flow cytometry. Fourteen nude mice were randomized into 2 groups (n=7) to receive subcutaneous injection of A549 cells with or without LASS2/TMSG1 overexpression on the back of the neck, and the cell proliferation in vivo was observed. The expression levels of p38 MAPK protein and p-p38 MAPK protein in the xenografts were detected with Western blotting. ELISA was used to detect the levels of ceramide and p38 MAPK protein in cultured A549 cell supernatants and the xenografts in nude mice.@*RESULTS@#Compared with the negative control cells, A549 cells with LASS2/TMSG1 overexpression had significantly lowered proliferation ability in vitro with increased early apoptosis rate (P < 0.05), and showed obvious growth inhibition after inoculation in nude mice(P < 0.05). Western blotting showed that in both cultured A549 cells and the xenografts in nude mice, LASS2/TMSG1 gene overexpression significantly increased the expression levels of p38 MAPK protein and p-p38 MAPK protein (P < 0.05); the results of ELISA also revealed significantly increased levels of ceramide and p38 MAPK protein in the cell supernatant andxenografts as well (P < 0.05).@*CONCLUSION@#Overexpression of LASS2/TMSG1 gene can significantly inhibit the proliferation and promote early apoptosis of human lung cancer A549 cells both in vitro and in vivo possibly by upregulating the expressions of ceramide and p38 MAPK protein to activate a signal transduction cascade.

Clinical efficacy of analysis of modified biliary-intestinal anastomosis by pancreaticoduodenec-tomy and influencing factors of postoperative biliary leakage / 中华消化外科杂志

Objective:To investigate the clinical efficacy of modified biliary-intestinal anasto-mosis by pancreaticoduodenectomy and influencing factors of postoperative biliary leakage.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopatholo-gical data of 165 patients with benign and malignant diseases around the ampullary who underwent pancreaticoduodenectomy in the Affiliated Hospital of Inner Mongolia Medical University from June 2014 to October 2020 were collected. There were 92 males and 73 females, aged (59±10)years. Of the 165 patients, 44 patients undergoing modified biliary-intestinal anastomosis within pancreatico-duodenectomy were divided into the modified group, and 121 patients undergoing traditional biliary-intestinal anastomosis within pancreaticoduodenectomy were divided into the traditional group. Observation indicators: (1) propensity score matching and comparison of general data of patients between the two groups after matching; (2) intraoperative and postoperative situations; (3) analysis of influencing factors of biliary leakage after pancreaticoduodenectomy. Propensity score matching was done by the 1:1 nearest neighbor matching method, with the caliper setting as 0.05. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Univariate analysis was conducted using the corresponding statistical methods based on data type. All indicators in univariate analysis were included in multivariate analysis. Multivariate analysis was conducted using the Logistic regression model. Results:(1) Propensity score matching and comparison of general data of patients between the two groups after matching. Of the 165 patients, 72 cases were successfully matched, including 36 cases in the modified group and 36 cases in the traditional group, respectively. The elimination of jaundice, preoperative reduction of jaundice and hypertension confounding bias ensured comparability between the two groups after propensity score matching. (2) Intraoperative and postoperative situations. All patients in the two groups underwent surgery successfully. The operation time, postoperative pathological type (lower bile duct cancer, pancreatic head cancer, pancreatic cystic tumor, chronic pancreatitis, duodenal cancer), time of no drainage fluid in the drainage tube around biliary-intestinal anastomosis were 371(270,545)minutes, 6, 12, 1, 2, 15, (12±7)days in patients of the modified group, versus 314(182,483) minutes, 13, 14, 1, 4, 4, (16±8)days in patients of the traditional group, showing significant differences in the above indicators between the two groups ( Z=-3.54, χ2=10.01, t=-2.34, P<0.05). Cases with postoperative grade A biliary leakage was 0 in patients of the modified group, versus 6 in patients of the traditional group, showing a significant difference between the two groups ( P<0.05). Cases with postoperative grade B biliary leakage, cases with postoperative grade B pancreatic fistula, cases with postoperative bleeding, cases with abdominal infection, cases with incision infection, cases with delayed gastric emptying, cases undergoing unplanned readmission were 1, 0, 1, 4, 1, 5, 1 in patients of the modified group, versus 0, 1, 2, 5, 2, 5, 2 in patients of the traditional group, showing no significant difference in the above indicators between the two groups ( P>0.05). Cases with postoperative grade A pancreatic fistula, cases with overall complications, cases with Clavien-Dindo grade Ⅰ-Ⅱ complications, cases with Clavien-Dindo grade Ⅲ-Ⅳ complications were 6, 12, 6, 6 in patients of the modified group, versus 7, 14, 8, 6 in patients of the traditional group, showing no significant difference in the above indicators between the two groups ( χ2=0.09, 0.24, 0.36, 0.00, P>0.05). None of patient in the two groups had postoperative grade C biliary leakage and postoperative grade C pancreatic fistula. (3) Analysis of influencing factors of biliary leakage after pancreaticoduodenectomy. Results of multivariate analysis showed that preoperative reduction of jaundice and traditional biliary-intestinal anastomosis were independent risk factors for biliary leakage after pancreaticoduodenectomy ( odds ratio=11.37, 12.27, 95% confidence interval as 1.76-73.35, 1.14-131.23, P<0.05). Conclusions:Compared with traditional biliary-intestinal anastomosis, modified biliary-intestinal anastomosis within pancreaticoduodenectomy is safe and feasible. Preoperative reduction of jaundice and traditional biliary-intestinal anastomosis are independent risk factors for biliary leakage after pancreaticoduodenectomy.

A multi-center study of biochemical and hotspot gene screening for neonatal genetic metabolic diseases / 中华实用儿科临床杂志

Objective:To investigate the efficiency of biochemical screening and hotspot gene screening in the detection of neonatal inherited metabolic diseases.Methods:This was a prospective multi-center study.The study was carried out on 21 442 neonatal samples collected from 12 hospitals in 10 provinces from November 2020 to November 2021.The results of biochemical screening and hotspot gene screening were analyzed jointly.Biochemical screening methods included glucose-6-phosphate dehydrogenase deficiency enzyme activity assay and neonatal tandem mass spectrometry.Genetic screening analysis involved 135 genes associated with 75 neonatal diseases.Results:Of all the 21 442 neonates enrolled in the study, 21 205 were subject to biochemical screening.A total of 813 cases were positive in the initial screening, and 0.45% of them (95 cases) were diagnosed after recall.All the 21 442 neonates underwent gene screening.About 168 positive cases were detected in the initial screening, and 0.73% (156 cases) of them were confirmed finally.Biochemical and genetic screening improved the detection sensitivity of such diseases as primary carnitine deficiency, neonatal intrahepatic cholestasis caused by citrin deficiency, and 2-methylbutyrylglycinemia.Moreover, biochemical and genetic screening enabled the detection of more diseases, including the common single-gene genetic diseases such as thalassemia and Wilson disease.Conclusions:In neonatal screening, the combination of biochemical screening and gene screening expands the number of diseases detected and improve screening efficiency.

Expression of TUBB4B in mouse primary spermatocyte GC-2 cells and its regulatory effect on NF-κB and MAPK signaling pathway / 南方医科大学学报

OBJECTIVE@#To explore the interaction between Tubulin beta 4B class IVb (TUBB4B) and Agtpbp1/cytosolic carboxypeptidase- like1 (CCP1) in mouse primary spermatocytes (GC-2 cells) and the role of TUBB4B in regulating the development of GC-2 cells.@*METHODS@#Lentiviral vectors were used to infect GC-2 cells to construct TUBB4B knockdown and negative control (NC-KD) cells. The stable cell lines with TUBB4B overexpression (Tubb4b-OE) and the negative control (NC-OE) cells were screened using purinomycin. RT-qPCR and Western blotting were used to verify successful cell modeling and explore the relationship between TUBB4B and CCP1 expressions in GC-2 cells. The effects of TUBB4B silencing and overexpression on the proliferation and cell cycle of GC-2 cells were evaluated using CCK8 assay and flow cytometry. The signaling pathway proteins showing significant changes in response to TUBB4B silencing or overexpression were identified using Western blotting and immunofluorescence assay and then labeled for verification at the cellular level.@*RESULTS@#Both TUBB4B silencing and overexpression in GC-2 cells caused consistent changes in the mRNA and protein expressions of CCP1 (P < 0.05). Similarly, TUBB4B expression also showed consistent changes at the mRNA and protein after CCP1 knockdown and restoration (P < 0.05). TUBB4B knockdown and overexpression had no significant effect on proliferation rate or cell cycle of GC-2 cells, but caused significant changes in the key proteins of the nuclear factor kappa-B (NF-κB) signaling pathway (p65 and p-p65) and the mitogen-activated protein kinase (MAPK) signaling pathway (ErK1/2 and p-Erk1/2) (P < 0.05); CCP1 knockdown induced significant changes in PolyE expression in GC-2 cells (P < 0.05).@*CONCLUSIONS@#TUBB4B and CCP1 interact via a mutual positive regulation mechanism in GC-2 cells. CCP-1 can deglutamize TUBB4B, and the latter is involved in the regulation of NF-κB and MAPK signaling pathways in primary spermatocytes.

Mechanism study of chlorogenic acid alleviating macrophage activation / 中国药房

OBJECTIVE To observe the effects of chlorogenic acid on the activation of macrophage induced by lipopolysaccharide （LPS）， and to explore the role of triggering receptors expressed on myeloid cells-2 （TREM2） in the action. METHODS To find a suitable LPS concentration， the cells were cultured with 1， 10 and 100 ng/mL LPS for 24 h. The level of interleukin 6 （IL-6） in the cell culture supernatant and protein expression of inducible nitric oxide synthase （iNOS） in the cells were detected. To search for a suitable chlorogenic acid concentration， the cells were divided into control group， LPS group and three chlorogenic acid （0.01， 0.1 and 1 μmol/L）+LPS groups. The levels of tumor necrosis factor α （TNF-α） and IL-1β in the cell culture supernatant， the protein expressions of iNOS and TREM2 in the cells and cell viability were detected. To observe the effects of TREM2 in chlorogenic acid alleviating macrophage activation， TREM2-small interfering RNA （TREM2-siRNA） was taken to intervene in TREM2 protein expression. The cells were divided into control group， LPS group， chlorogenic acid+LPS group， TREM2-siRNA+chlorogenic acid+LPS group and SC-siRNA+chlorogenic acid+LPS group. After 24 h incubation， the levels of TNF- α and IL-1β in the cell culture supernatant and protein expressions of TREM2， iNOS and nuclear factor κB p65 （NF-κB p65） in the cells were detected. RESULTS 10 ng/mL LPS promoted IL-6 release and increased iNOS protein expression， and 10 ng/mL LPS was taken in the next experiments. Compared with the LPS group， 0.1 μmol/L chlorogenic acid decreased TNF-α jiaji1981@126.com and IL-1β levels， and down-regulated iNOS expression，meanwhile increased TREM2 expression without effect on cell viability， and 0.1 μmol/L chlorogenic acid was taken in the next experiments. Compared with the control group， the protein expressions of iNOS and NF- κB p65 in the LPS group were significantly increased （P＜0.05）； compared with the LPS group， the protein expressions of iNOS and NF- κB p65 in the chlorogenic acid+LPS group were significantly decreased， the protein expressions of TREM2 was significantly increased （P＜ 0.05）； compared with the chlorogenic acid+LPS group， the protein expressions of iNOS and NF-κB p65 of TREM2-siRNA+ chlorogenic acid+LPS group were significantly increased， the protein expressions of TREM2 was significantly decreased （P＜0.05）. TREM2-siRNA could significantly reverse the above effects of chlorogenic acid， while SC-siRNA did not significantly affect the above anti-inflammatory effects of chlorogenic acid. CONCLUSIONS Chlorogenic acid can inhibit the LPS-induced macrophage activation， and its anti-inflammatory may be mediated by TREM2 protein.

Differential Expression of Long Non-coding RNA Cancer Susceptibility Candidate 2 and Imprinted Gene H19 in Extrahepatic Cholangiocarcinoma / 中国医学科学院学报

Objective To investigate the expression and the potential roles of long non-coding RNA(lncRNA)cancer susceptibility candidate 2(CASC2)and imprinted gene H19 in extrahepatic cholangiocarcinoma(ECC). Methods Four samples from patients with ECC were collected for high-throughput sequencing which was conducted to reveal the transcriptomic profiles of lncRNA CASC2 and H19.Bioinformatics tools were employed to predict the potential roles of the two genes.Another 22 ECC tissue samples and the cholangiocarcinoma cell lines(RBE,QBC939,HuH-28,and HuCCT1)with different degrees of differentiation were selected for validation.The para-carcinoma tissue and normal human intrahepatic biliary epithelial cell(HIBEC)were used as the control groups.The expression levels of lncRNA CASC2 and H19 in carcinoma tissue,para-carcinoma tissue,and cell lines were determined by real-time quantitative polymerase chain reaction(qRT-PCR).The correlation analysis was carried out for the clinical indicators of patients with the expression levels of the target genes. Results The two target genes showed significantly different expression between carcinoma tissue and para-carcinoma tissue(all P<0.05).Specifically,CASC2 had higher expression level in the carcinoma tissue than in the para-carcinoma tissue(t=1.262,P=0.025),whereas the expression of H19 showed an opposite trend(t=1.285,P=0.005).The expression levels of CASC2 in QBC939(t=8.114,P=0.015)and HuH-28(t=9.202,P=0.012)cells were significantly higher than that in the control group.The expression levels of H19 were significantly lower in RBE(t=-10.244,P<0.001),QBC939(t=-10.476,P<0.001),HuH-28(t=-19.798,P<0.001),and HuCCT1(t=-16.193,P=0.004)cells than in the control group.Bioinformatics analysis showed that CASC2 was mainly involved in the metabolic process and H19 in the development of multicellular organisms.Both CASC2 and H19 were related to catalytic activity.The expression level of lncRNA CASC2 was correlated with pathological differentiation(χ 2=6.222,P=0.022)and lymph node metastasis(χ2=5.455,P=0.020),and that of lncRNA H19 with pathological differentiation(χ2=1.174,P=0.029)and tumor size(χ2=-0.507,P=0.037). Conclusions In the case of ECC,lncRNA CASC2 and H19 have transcription disorders.lncRNA CASC2 is generally up-regulated in the carcinoma tissue,while H19 is down-regulated.Both genes have the potential to become new molecular markers for ECC.

Avascular necrosis of the metacarpal head: a case report and literature review / 中华骨科杂志

Osteonecrosis is the most common disorder in femoral head and scaphoid. However, avascular necrosis (AVN) in metacarpal head is a rare disease. In the present study, a 14-year-old male patient complained of right-hand pain with a limited range of motion for one month. Physical examination showed that the active flexion was 70° and the extension was limited to 30° of the metacarpophalangeal (MCP) joints. The grip strength decreased to 60% of the contralateral hand. X-rays demonstrated flattening and sclerotic changes in the long finger and ring finger of metacarpal head. CT scanning indicated cystic, osteochondral defects and sclerotic changes in metacarpal head. Imaging examination further confirmed the diagnosis of AVN in the long metacarpal finger and ring finger. After conservative treatments including splint immobilization, non-steroidal anti-inflammatory drugs (NSAIDs) and physiotherapy, the local pain symptoms of the MCP joint gradually disappeared. The range of motion of MCP joint returned to normal with the grip strength score as 105% of the contralateral hand. MRI confirmed excellent remodeling and regeneration in the metacarpal head at two years later. The clinical characteristics, diagnosis and treatments of AVN of metacarpal head were reviewed. Although radiograph examination is commonly used, early-stage osteonecrosis of the metacarpal head should be confirmed by MRI. Given the rarity of this disorder, there is no consensus on the treatments. Metacarpal necrosis is the more common disorder in adolescent patients with a history of trauma. Considering the potential of bone remodeling, juveniles with metacarpal head necrosis could recover by conservative treatments.

Quantitative analysis methods of white matter microstructure based on diffusion tensor imaging and its application in chronic pain research / 生理学报

As the two essential components, the white matter and gray matter compose the central nervous system of the brain. Widely known that axons of neurons mainly form the white matter, and these formed nerve fibers are responsible for transmitting information among various brain regions to achieve the coordinated operation of the entire brain. Early research on the white matter could only be done by dissecting living animals or human cadavers, until Basser et al. proposed diffusion tensor imaging (DTI) technology in 1994, which could detect the diffusion characteristics of water in the brain in vivo noninvasively. Accordingly, this technology could be applied to investigate the diffusion movement of water in white matter to obtain the information of direction and micro-anatomy of white matter fiber bundles. With the advancement on the display and analysis of the anatomical structure of white matter fiber bundles, the exploration of microscopic pathological changes, and the assistance of clinical diagnosis and neurophysiological research, DTI technology has become one of the most popular topics in brain science research. Chronic pain refers to pain lasting more than three months, which not only seriously affects the patient's physical and social functions, but also dramatically reduces the quality of life. It was reported that long-term pain stimulation might cause pathological remodeling of the central nervous system, and abnormalities in white matter were found in imaging examinations of patients with chronic pain. This review introduces the quantitative analysis methods of white matter fiber bundle microstructure based on DTI and its application in chronic pain, and further discusses the application value of DTI technology on clinical research of chronic pain.

CT and MRI features of acinar cell carcinoma of the pancreas / 中华消化外科杂志

Objective:To summarize the computed tomography (CT) and magnetic resonance imaging (MRI) features of acinar cell carcinoma of the pancreas (ACCP).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 21 patients with ACCP who were admitted to the Affiliated Hospital of Inner Mongolia Medical University from January 2015 to December 2019 were collected. There were 5 males and 16 females, aged (57±9)years, with a range from 41 to 74 years. Patients underwent CT and MRI examinations. Observation indicators: (1) imaging examination; (2) imaging features on CT; (3) imaging features on MRI; (4) pathological examination and immunohistochemistry staining; (5) treatment and follow-up. Follow-up using outpatient examination and telephone interview was conducted at 1, 3, 6 months after discharge and once every 6 months thereafter to detect survival of patients up to December 2019. Measurement data with normal distribution were represented as Mean± SD. Count data were described as absolute numbers. Results:(1) Imaging examination. Of the 21 patients, 7 underwent single CT examination, 11 underwent MRI examination, and 3 underwent both CT and MRI examinations. ① Tumor shape: all the 21 patients had single tumor, including 17 showing round or quasi-round shape, and 4 showing irregular clumps. ② Tumor location: of the 21 patients, 6 had tumor located at pancreatic head, 2 had tumor located at pancreatic head and body, 2 had tumor located at pancreatic body, 4 had tumor located at pancreatic body and tail, 4 had tumor located at pancreatic tail, and 3 had had tumor located at ampulla. ③ The maximum tumor diameter was (43±29)mm, with a range from 11 to 129 mm. ④ Adjacent organ invasion: 10 of the 21 patients had invasion of adjacent organ, including 2 with invasion of stomach, spleen and left adrenal gland invasion, 4 with invasion of duodenum, 3 with invasion of duodenum and common bile duct, 1 with invasion of spleen. ⑤ Vascular invasion: 12 patients had invasion of splenic artery or splenic vein, including 1 combined with invasion of both common hepatic artery and superior mesenteric vein, 1 combined with invasion of celiac root. ⑥ Pancreatic and bile duct invasion: 8 patients had pancreatic and bile duct dilation, including 4 with bile duct and upper pancreatic duct dilation, and 4 with pancreatic duct dilation. ⑦ Lymph node metastasis: 2 patients had perineoplastic lymph node enlargement. ⑧ Other conditions: 7 patients had tumor center with cystic necrosis. Four patients had atrophy pancreatic parenchyma. Two patients had splenic vein tumor thrombosis. Two patients had cysts. One patient had multiple liver metastases. (2) Imaging features on CT. ① The solid part was dominant in the main body of the 10 patients undergoing CT examination, demostrating equal density, of which 3 cases had clear boundaries, 2 cases had pseudocapsule around the lesion, and 5 cases had low-density necrotic area in the center of lesion. ② In arterial phase of CT examination, the solid part of tumor had a lower enhancement compared with the normal pancreatic tissues in 7 patients, while the solid part of tumor had a high enhancement compared with the normal pancreatic tissues in 3 patients. ③ In delayed phase of CT examination, the tumor density was slightly lower than or equal to density of normal pancreatic parenchyma in 7 patients, showing slightly progressive enhancement, while the tumor density was slightly higher than or equal to density of normal pancreatic parenchyma in 3 patients. (3) Imaging features on MRI. ① MRI plain scan of 14 patients showed that 8 patients demostrated slightly longer T2 and slightly longer T1 signals in lesions, while 6 patients demostrated mixed signals dominated by long T2 and equal T1 signals. The area of cystic necrosis was observed in lesions of 4 patients and was not observed in 10 patients. No antiphase signal reduction was observed in the 14 patients. ② MRI dynamic enhanced scan of 12 patients showed that 11 patients presented mild progressive enhancement in lesions and 1 patient presented obvious confounding enhancement and clearance in the delayed phase. Compared with adjacent normal pancreatic parenchyma, diffused weighted imaging showed high signals in 6 cases, slightly high signals in 6 cases, and high signal halo in 2 cases. The apparent diffusion coefficient in 14 lesions was (1.22±0.14)×10 -3 mm 2/s. (4) Pathological examination and immunohistochemistry staining. Results of pathological examination in the 21 patients: acinic cell carcinoma, mixed ductal-acinic cell carcinoma, acinar-endocrine carcinoma, and atypical hyperplasia inacinus were detected in 14, 5, 1, and 1 patients, respectively. Of the 21 patients, 10 had invasion of adjacent organ, 3 had invasion of bile duct, 2 had invasion of lymph node. Results of immunohistochemistry staining in 17 patients: 17 patients had proliferation index of Ki-67 as 1%-80%; 10 out of 16 patients were positive for synaptophysin; 6 out of 16 patients were positive for CD56 protein; 2 out of 14 patients were positive for Chromogranin A; 12 out of 13 patients were positive for α-antitrypsin; 9 out of 11 patients were positive for cytokeratin; 8 patients were positive for β-catenin; 2 patients were positive for B lymphoma-10 protein. (5) Treatment and follow-up. Of the 21 patients, 10 cases underwent pancreatico-duodenectomy, 6 cases underwent pancreatic body and tail pancreatectomy combined with splenectomy, 2 cases underwent pancreatic body and tail pancreatectomy, 1 case underwent pancreatic tail tumor enucleation, 1 case underwent liver metastasis resection, and 1 case underwent ultrasound-guided pancreatic lesion puncture biopsy. All the 21 patients were followed up for (30±16)years, with a range from 2 to 52 months. There were 13 patient surviving and 8 cases of death. They had survived for (19±13)months, with a range from 2 to 35 months. Conclusions:The CT and MRI enhanced scan of ACCP showed slightly progressive enhancement, with cystic necrosis seen in the center and high signals in diffused weighted imaging. Dilation of bile duct and pancreatic duct is common in patients with pancreatic head tumors, and invasion of splenic artery and vein is common in pancreatic body and tail tumors. Calcification and cyst are rare and lesions of pancreatic head and body cause atrophy in pancreatic tail.

Safety and efficacy of modified liquid isolation method in microwave ablation in treatment of secondary hyperparathyroidism / 中国介入影像与治疗学

Objective: To observe the safety and efficacy of modified liquid isolation method during ultrasound-guided microwave ablation in treatment of secondary hyperparathyroidism (SHPT). Methods: Clinical data of 82 SHPT patients who underwent ultrasound-guided microwave ablation were retrospectively analyzed. Among them, 41 cases adopted improved liquid isolation method (observation group), while the other 41 cases received traditional liquid isolation method (control group). Unit volume ablation time, serum intact parathyroid hormone (iPTH), iPTH change rate and serum calcium before and 7 days after operation, the incidence and recovery rate of recurrent laryngeal nerve injury during and 6 months after operation were compared between the two groups. Results: There was no significant difference of unit volume ablation time, serum iPTH, iPTH change rate nor serum calcium level before and 7 days after operation between the two groups (all P>0.05). The incidence of recurrent laryngeal nerve injury in observation group during and 6 months after operation was 14.63% (6/41), lower than that of control group (43.90%[18/41], P=0.004). Six months after operation, all 6 patients (6/6, 100%) with recurrent laryngeal nerve injury in observation group and 13 patients (13/18, 72.22%) in control group recovered (P=0.003). Conclusion: Modified liquid isolation microwave ablation can effectively treat SHPT and reduce recurrent laryngeal nerve injury.

Clinical application value of modified invagination for pancreaticojejunostomy in pancreaticoduodenectomy / 中华消化外科杂志

Objective To explore the clinical application value of modified invagination for pancreaticojejunostomy in pancreaticoduodenectomy (PD).Methods The retrospective cohort study was conducted.The clinicopathological data of 39 patients who underwent PD in the Affiliated Hospital of Inner Mongolia Medical University from January 2014 to December 2017 were collected.There were 26 males and 13 females,aged (60±7)years,with a range of 41-75 years.All the 39 patients underwent PD,using Child method to reconstruct digestive tract.Of 39 patients,19 undergoing modified invagination for pancreaticojejunostomy and 20 undergoing mucosa-to-mucosa end-to-side pancreaticojejunostomy were allocated to innovative group and traditional group,respectively.Observation indicators:(1)surgical situations;(2) postoperative complications;(3) follow-up.Follow-up was performed by outpatient examination and telephone interview to detect postoperative tumor recurrence,main pancreatic duct dilatation,survival,and discomfort (abdominal pain,bloating,indigestion,etc.) of patients up to October 2018.Measurement data with normal distribution were represented as Mean±SD,and comparison between groups was analyzed by t test.Measurement data with skewed distribution were represented as M (P25,P75) or M (range),and comparison between groups was analyzed by Mann Whitney U test.Count data were expressed as absolute numbers,and comparison between groups was analyzed by chi-square test or Fisher exact probability.Results (1) Surgical situations:operation time,volume of intraoperative blood loss,cases with soft pancreas or hard pancreas (pancreatic texture),pancreatic duct diameter,time of pancreatic duct removal,cases using somatostatin,and duration of postoperative hospital stay of the innovative group were (342±47) minutes,400 mL (300 mL,400 mL),10,9,3.1 cm (2.9 cm,3.4 cm),37 days (32 days,63 days),17,18 days (15 days,22 days),respectively,versus (392±95)minutes,400 mL (300 mL,525 mL),6,14,3.6 cm (2.6 cm,4.2 cm),43 days (34 days,49 days),18,and 24 days (15 days,27days) of the traditional group;there was no significant difference in the volume of intraoperative blood loss,cases with soft pancreas or hard pancreas (pancreatic texture),pancreatic duct diameter,time of pancreatic duct removal,cases using somatostatin,and duration of postoperative hospital stay between the two groups (Z=-0.775,x2 =2.063,Z=-1.155,Z=-0.295,x2 =0.003,Z=-1.286,P＞0.05);but a significant difference in operation time between the two groups (t =-2.114,P＜0.05).(2) Postoperative complications:6 patients in the innovative group had complications,including 1 of delayed gastric emptying,1 of wound infection,1 of pulmonary infection,1 of acute respiratory failure,1 of perihepatic effusion,and 3 of grade A pancreatic leakage;11 patients in the traditional group had postoperative complications,including 1 of bile leakage,2 of delayed gastric emptying,4 of abdominal infection,4 of wound infection,2 of pulmonary infection,1 of ascites,1 of abdominal hemorrhage,1 of pleural effusion,2 of grade A pancreatic leakage,5 of grade B and C pancreatic leakage;the same patient had multiple complications.There was no significant difference in postoperative complications between the two groups (x2=2.174,P＞0.05),but there was a significant difference in postoperative grade B and C pancreatic leakage between the two groups (P＜O.05).Patients with postoperative complications were improved after symptomatic support treatment,and no patient died during the perioperative period.(3) Follow-up:of the 39 patients,33 (18 in the innovation group and 15 in the traditional group) were followed up for 3-57 months,with a median follow-up time of 17 months.Of the 18 patients receiving follow-up in the innovative group,5 died of tumor recurrence and metastasis,with a survival time of 5-24 months,1 had tumor recurrence at 34 months after operation,1 had main pancreatic duct dilatation and intermittent abdominal pain and abdominal distension,5 had indigestion,1 had back pain,and 5 had good recovery.Of 15 patients receiving follow-up in the traditional group,10 died of tumor recurrence and metastasis,with a survival time of 3-57 months,2 had main pancreatic duct dilatation and intermittent abdominal pain and abdominal distension,2 had indigestion,1 had good recovery.Conclusion Compared with the traditional mucosa-to-mucosa end-to-side pancreaticojejunostomy,modified invagination for pancreaticojejunostomy in the PD is safe and feasible,which can simplify the operation,reduce the requirements for the operator's operation skills,shorten the operation time,and reduce incidence of postoperative grade B and C pancreatic leakage.

Computed tomography and magnetic resonance imaging features of regional portal hypertension / 中华消化外科杂志

Objective To investigate the features of computed tomography (CT) and magnetic resonance imaging (MRI) of regional portal hypertension (RPH).Methods The retrospective cohort study was conducted.The clinicopathological data of 31 patients with PHR in the RPH group and 31 patients with liver cirrhotic portal hypertension (CPH) in the CPH group who were admitted to the Affiliated Hospital of Inner Mongolia Medical University between February 2014 and February 2018 were collected.Etiologies of patients in the RPH group included 21 of chronic pancreatitis complicated with pancreatic pseudocyst,5 of carcinoma of pancreatic body and tail,1 of solid pseudopapillary tumor of the pancreas,1 of pancreatic serotls cystoadenoma,1 of gastric stromal tumor,1 of retroperitoneal metastatic tumor and 1 of left renal carcinoma.Etiologies of patients in the CPH group included 27 of liver cirrhosis after viral hepatitis type B (4 complicated with liver metastasis),3 of alcoholic cirrhosis and 1 of cholestatic cirrhosis.All the patients underwent CT and MRI examinations.Patients in the RPH group were mainly treated the primary diseases and patients in the CPH group were decreased portal vein pressure.Observation indicators:(1) imaging features of patients in the two groups;(2) treatment and follow-up situations.Follow-up using outpatient examination and telephone interview were performed to detect management of portal hypertension after treatment up to February 2018.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed by the t test.Measurement data were represented as M (range) and comparison between groups was analyzed by the Mann-Whitney rank sum test.Count data were compared with chi-square test.Results (1) Imaging features of patients in the two groups:of 31 patients in the RPH group,12 underwent CT examination,2 underwent MRI examination,and 17 underwent CT combine with MRI examination.Of 31 patients in the CPH group,12 underwent CT examination and 19 underwent CT combined with MRI examination.The number of patients with varices in the gastric fundus,the number of patients with combined esophageal varices,the number of perigastric varices,diameter of main portal vein,diameter of splenic vein,liver volume,splenic volume,hepatosplenic volume ratio were 11,1,49,(13.9±2.9) mm,(12.0±2.8) mm,1 383 cm3 (range,1 005-1 637 cm3),271 cm3(range,199-311 cm3) and 5.5±2.0 in the RHP group and 24,21,33,(16.3±1.7)mm,(10.5±3.2)mm,1 087 cm3(range,916-1 536 cm3),603 cm3(range,415-869 cm3) and 2.2±0.9 in the CHP group,with statistically significant differences in the number of patients with varices in the gastric fundus,the number of patients with combined esophageal varices,the number of perigastric varices,diameter of main portal vein,splenic volume,hepatosplenic volume ratio between the two groups (x2=11.088,28.182,8.940,t=4.430,Z=6.205,t=8.544,P＜0.05) and with no statistically significant differences in the diameter of splenic vein and liver volume between the two groups (t=1.974,Z=1.162,P＜0.05).Of 31 patients in the RPH group,2 with pancreatic pseudocyst were misdiagnosed as pancreatic cancer and 29 were diagnosed accurately by imaging examinations.Of 31 patients in the CPH group,3 with liver metastasis were undetected by CT examination and the other 28 were diagnosed accurately by imaging examinations.Splenic vein occlusion,severe splenic vein stenosis,moderate splenic vein stenosis and mild splenic vein stenosis were detected in 2,17,10 and 2 of 31 patients in the RHP group.All the 31 patients in the CHP group mainly had dilation in splenic veins,with no clear stenosis.(2) Treatment and follow-up situations:patients in the two groups were followed up for 6-48 months,with a median time of 21 months.Of 21 patients with chronic pancreatitis complicated with pancreatic pseudocyst in the RPH group,7 underwent pancreatic pseudocyst puncture and drainage,6 of them had poor control on portal hypertension and 1 had moderate control;4 underwent pancreaticoenteric drainage,1 of them underwent pancreaticojejunostomy 4 years later and 3 of them had good control on portal hypertension;3 undergoing splenectomy combined with perigastrectomy had good control on portal hypertension;7 undergoing conservative treatment had good control on portal hypertension.Of 5 patients with carcinoma of pancreatic body and tail in the RHP group,2 undergoing distal pancreatectomy combined with splenectomy had good control on portal hypertension and 3 undergoing non-operative combined therapy died of primary disease one year later.One,1 and 1 patient with solid pseudopapillary tumor of the pancreas,pancreatic serotls cystoadenoma and gastric stromal tumor respectively in the RHP group underwent relative surgical treatments and had good control on portal hypertension.One and 1 patient with retroperitoneal metastatic tumor and left renal carcinoma respectively in the RHP group underwent non-operative combined therapy and had good control on portal hypertension.All the 31 patients in the CHP group were mainly treated with protection of liver function,8 of them were encounted with medusa head,7 with upper gastrointestinal rehemorrhage within one year,5 with subcutaneous varicose vein of abdominal wall,3 with continuing increase of spleen volume and 8 had good control on portal hypertension.Conclusions RHP are existed in pancreatic,splenic or peritoneal diseases,especially the pancreatic primary diseases.The main imaging features of RHP include isolated gastric varices,perigastric varices and splenic vein occlusion without normal main portal vein and liver function.Surgical resection of primary tumor and reasonable splenectomy are effective therapy.

New mutation of ACADVL-related very long chain acyl-CoA dehydrogenase deficiency / 中华实用儿科临床杂志

Objective According to the clinical and gene mutation characteristics of ACADVL-related very long chain acyl-CoA dehydrogenase deficiency(VLCADD),the types that contribute to the gene mutation of ACADVL were summarized.Methods By analyzing clinical,laboratory and genetic data of 1 case with ACADVL-related very long chain acyl-CoA dehydrogenase deficiency diagnosed from Department of Pediatrics,the Affiliated Hospital of Inner Mongolia Medical University in August 2016,based on the agreement signed by both the litde patient's parents and the hospital,plus the high-throughput sequencing analysis and PCR sequencing test for the 2 generation genes,some presented mutation sites were analyzed and concluded,in addition to taking "ACADVL" as key words to search the databases from CNKI,Wanfang(updated in 2016) as well as PubMed and related documents from On-line Mendal Inheritant databases of Man (OMIM) and HGMD.Results Through physical examination,VLCADD was diagnosed.After being given Levocamitine and the diet likemedium-chain fatty acid food for a week,the metabolism returned to normal.Tracking him for 3 months,his hepatitis obviously rebounded,within the reach of 3 cm under the right rib and 1 cm under the xiphoid.The exome sequencing study (trios) was identified the novel heterozygous mutation according to the statistics below A CAD VL (N M_000018.3) Exon7:c.608 C ＞ T;p.(Pro203 Leu) (heterozygous) and A CAD VL (NM _000018.3) Exon18:c.1748C ＞ T;p.(Ser583Leu) (heterozygous) in ACADVL.Relevant literature reported suggest these two mutations from both the parents are pathogenic genes,which can account for the reason why the boy got ill.However,these two mutations had not been reported in ACADVL-related VLCADD so far.Up to now,73 types of mutations from documents index were related to the VLCADD,but the clinical case included 75 kinds of gene mutations.Conclusions The apparent symptoms of the boy with the gene mutation were reflected in abnormal heart rates,hepatomegaly and hypoglycemia.VLCADD was diagnosed through genetic testing,and systematic treatment can partly control the development of the disease.In conclusion,the findings (exon 7 and 18) show that according to the genetic tests,disease-causing genes from both parents are new mutations of ACADVL and they are pathogenic.

Feasibility of body surface electrodes instead of multipair esophageal electrodes for assessment of neural re-spiratory drive in COPD patients / 实用医学杂志

Objective To analyze the feasibility of body surface electrodes instead of multipair esophageal electrodes for the evaluation of neural respiratory drive in patients with COPD. Methods Diaphragm electromyo-gram(EMG)from body surface electrodes and multipair esophageal electrodes,was recorded in 29 patients with stable COPD recruited from outpatient clinic. Changes of neural respiratory drive of two kinds of electrodes during resting and maximal isocapnic ventilation (MIV) were observed before and after inhalation of bronchodilators. Results Ventilation significantly improved ,RMS-sur and RMS-eso significantly decreased after the inhalation of bronchodilators during resting and MIV. RMS-sur and RMS-eso were significantly correlated(r=0.660,P<0.01). Conclusion EMG from the surface electrodes may be a useful and noninvasive technique to evaluate neural respi-ratory drive in patients with COPD.

Effect of maternal separation stress on behavior of neonatal rd mice / 中国比较医学杂志

Objective To evaluate the effect of maternal separation stress on the behavior of neonatal rd mice.Methods Neonatal rd mice were divided into maternal separation (MS) group (n=9) and control group (n=9).MS-stress was induced in the MS group by 4-hour-separation per day for 28 days.Open field test,elevated plus maze test,forced swim test and tail suspension test were used to evaluate the anxiety-like and depression-like behavior of the neonatal rd mice.Results The stay time and distance travelled of MS group in the central zone were 0.88％ and 28.17±5.65 cm,respectively,significantly shorter than that of the control group (2.61％,109.9±9.79 cm.P =0.04,P =0.001).Compared with the control group,the stay time in open arms of the MS group was significantly decreased (P<0.01),while the immobility time in forced swim test and tail suspension test of the MS group were 126.5±10.22 s and 21.56±6.83 s,significantly longer than that of the control group (77.75±16.83 s,P =0.02,7.37±3.22 s,P =0.03).Conclutions The 28-day maternal separation stress can significantly increase the anxiety-like and depression-like behavior in neonatal rd mice.

Exploration and optimization of processes for key intermediates of phosphorodiamidate morpholino oligonucleotides / 军事医学

Objective To explore and optimize the processes for synthesis of key intermediates of phosphorodiamidate morpholino oligonucleotides-7'-hydroxy-N-trityl morpholino nucleoside monomer in order to contribute to the research of phosphorodiamidate morpholino oligonucleotides antisense nucleotides.Methods With N-benzoylcytidine,guanosine and 5-methyluridine as starting materials,the ribose was modified to morpholino and the key chemical groups were protected to obtain 7'-hydroxy-N-trityl morpholino nucleoside monomer.Results Compounds N4-benzoyl-7'-hydroxy-N-trityl morpholinocytidine,N2-benzoyl-7'-hydroxy-N-trityl morpholinoguanosine and 7'-hydroxy-N-trityl morpholinothymidine were synthesized.The synthetic processes were optimized as well.The structures of all the intermediates and title compounds were characterized.Conclusion The synthetic processes of 7'-hydroxy-N-trityl morpholino nucleoside monomers have been optimized,which can be employed to prepare title compounds on a large scale.

Relationship between subtypes of T follicular helper cells and unexplained recurrent spontaneous abortion / 上海交通大学学报（医学版）

Objective · To discuss the relationship between unexplained recurrent spontaneous abortion (URSA) and T follicular helper (Tfh) cell subtypes. Methods · Twenty-eight normal early pregnancy women, who had undergone induced abortion, were taken as control, and 28 patients with URSA were enrolled in the abortion group. The mononuclear cells in peripheral blood and decidual tissues were separated in the two groups, and CXCR3+CCR6- Tfh cells, CXCR3-CCR6- Tfh cells, CXCR3-CCR6+ Tfh cells and B cells were tested by flow cytometry. Results · The decidual CXCR3-CCR6- Tfh cells significantly increased in the abortion group compared with control group (P=0.015). And there was a strong association between the decidual CXCR3-CCR6- Tfh cells and B cells in URSA patients (R2=0.779, P=0.025). Conclusion · The up-regulation of decidual CXCR3-CCR6- Tfh cells in early pregnancy women may be related with the occurrence of URSA.

Pretreatment with Pam3 CSK4 decreases inflammation in renal tissue from mice with systemic MRSA infection / 中国免疫学杂志

Objective:To observe whether pretreatment with Pam3CSK4,a TLR2 agonist,could decrease the inflammation response in kidney from mice with systemic MRSA infection,and to investigate the mechanism of the attenuation of inflammation with Pam3CSK4 pretreatment. Methods:BALB/c mice were pretreated with Pam3CSK4 (10 μg/100 μl/each mouse) or PBS via tail vein once daily for two consecutive days. All mice were infected with live MRSA (ATCC43300) at 2×107 CFU/each mouse (via tail vein) 24 h after the second treatment. The levels of cytokines in kidney were measured by ELISA and real-time PCR,respectively. The relative expression of TLR2,IRAKs etc. were detected by real-time PCR. Western blot was performed to detect the phosphorylation of NF-κB, the expression of IRAK-M and A20,respectively. Results:The level of TNF-α,IL-6,IL-1β,CCL3 and IFN-γ in renal tissue from mice pretreated with Pam3CSK4 was decreased significantly compared with that from PBS-treated mice,respectively. Pam3CSK4 pretreatment down-regulated the relative expression of TLR2, inhibited the expression of IRAK-1 and the phosphorylation of NF-κB post infection. The expression of IRAK-M,one of the negative regulators in TLRs signaling pathway was increased significantly in renal tissue from Pam3CSK4-treated mice post infection. Conclusion:Pam3CSK4 pretreatment attenuated the inflammation response in kidney from mice with systemic MRSA infection,and these attenuation is related with up-regulation of IRAK-M.

Relationship between subtypes of T follicular helper cells and unexplained recurrent spontaneous abortion / 上海交通大学学报（医学版）

Objective · To discuss the relationship between unexplained recurrent spontaneous abortion (URSA) and T follicular helper (Tfh) cell subtypes. Methods · Twenty-eight normal early pregnancy women, who had undergone induced abortion, were taken as control, and 28 patients with URSA were enrolled in the abortion group. The mononuclear cells in peripheral blood and decidual tissues were separated in the two groups, and CXCR3+CCR6- Tfh cells, CXCR3-CCR6- Tfh cells, CXCR3-CCR6+ Tfh cells and B cells were tested by flow cytometry. Results · The decidual CXCR3-CCR6- Tfh cells significantly increased in the abortion group compared with control group (P=0.015). And there was a strong association between the decidual CXCR3-CCR6- Tfh cells and B cells in URSA patients (R2=0.779, P=0.025). Conclusion · The up-regulation of decidual CXCR3-CCR6- Tfh cells in early pregnancy women may be related with the occurrence of URSA.

Pretreatment with Pam3 CSK4 decreases inflammation in renal tissue from mice with systemic MRSA infection / 中国免疫学杂志

Objective:To observe whether pretreatment with Pam3CSK4,a TLR2 agonist,could decrease the inflammation response in kidney from mice with systemic MRSA infection,and to investigate the mechanism of the attenuation of inflammation with Pam3CSK4 pretreatment. Methods:BALB/c mice were pretreated with Pam3CSK4 (10 μg/100 μl/each mouse) or PBS via tail vein once daily for two consecutive days. All mice were infected with live MRSA (ATCC43300) at 2×107 CFU/each mouse (via tail vein) 24 h after the second treatment. The levels of cytokines in kidney were measured by ELISA and real-time PCR,respectively. The relative expression of TLR2,IRAKs etc. were detected by real-time PCR. Western blot was performed to detect the phosphorylation of NF-κB, the expression of IRAK-M and A20,respectively. Results:The level of TNF-α,IL-6,IL-1β,CCL3 and IFN-γ in renal tissue from mice pretreated with Pam3CSK4 was decreased significantly compared with that from PBS-treated mice,respectively. Pam3CSK4 pretreatment down-regulated the relative expression of TLR2, inhibited the expression of IRAK-1 and the phosphorylation of NF-κB post infection. The expression of IRAK-M,one of the negative regulators in TLRs signaling pathway was increased significantly in renal tissue from Pam3CSK4-treated mice post infection. Conclusion:Pam3CSK4 pretreatment attenuated the inflammation response in kidney from mice with systemic MRSA infection,and these attenuation is related with up-regulation of IRAK-M.