[摘 要] 目的：探究双特异性磷酸酶26（DUSP26）在肺腺癌（LUAD）A549细胞增殖、迁移和侵袭中的作用及其分子机制。方法：检索肿瘤数据库GEPIA2网站DUSP26表达数据，分析DUSP26在LUAD患者和正常人肺组织中的表达差异。收集2022年10月至2023年10月期间萍乡市人民医院手术切除的12例LUAD组织和癌旁组织标本，通过免疫组织化学（IHC）和WB法检测DUSP26在LUAD组织和癌旁组织之间的表达差异；通过WB法检测DUSP26在4种LUAD细胞（A549、SK-LU-1、Calu-3、H1299）和2种正常支气管上皮细胞（BEAS-2B、HBEC）中的表达差异。利用慢病毒转染细胞的方法构建稳定过表达DUSP26（DUSP26-OE）及阴性对照（DUSP26-OENC）的A549细胞，通过克隆形成、划痕愈合实验、Transwell实验分别检测DUSP26过表达对细胞增殖、迁移及侵袭能力的影响，WB法检测各组细胞中TGF-β1/SMAD2/3通路、EMT相关蛋白的表达水平，细胞免疫荧光法检测细胞中Ki-67、cyclin D1表达水平。加入TGF-β1重组蛋白进行回复实验。构建A549细胞裸鼠荷瘤模型，观察DUSP26过表达对移植瘤体内生长的影响，WB法检测移植瘤组织中TGF-β1/SMAD2/3通路、EMT相关蛋白表达水平，免疫荧光染色法检测移植瘤组织中Ki-67、cyclin D1表达水平。结果：DUSP26在LUAD组织和细胞中均呈低表达（P < 0.05或P < 0.01或P < 0.001或P < 0.000 1）。与DUSP26-OENC组相比，DUSP26-OE组A549细胞的增殖、迁移和侵袭能力均显著降低（P < 0.01或P < 0.001），TGF-β1、p-SMAD2/3、vimentin、N-cadherin、snail、Ki-67、cyclin D1表达均降低（P < 0.01或P < 0.001或P < 0.000 1），E-cadherin表达升高（P < 0.000 1）。加入5 ng/mL TGF-β1重组蛋白后，可部分逆转在体外实验中由DUSP26过表达导致的结果。成功构建裸鼠A549细胞荷瘤模型，DUSP26-OE组裸鼠移植瘤生长速度缓慢，体积和质量均减小（均P < 0.001），移植瘤组织中TGF-β1、p-SMAD2/3、vimentin、N-cadherin、snail、Ki-67、cyclin D1表达均降低（P < 0.01或P < 0.001），E-cadherin表达升高（P < 0.000 1）。结论：DUSP26在LUAD组织和细胞中均呈低表达状态，上调DUSP26的表达水平能够通过抑制TGF-β1/SMAD2/3信号通路抑制A549细胞的增殖、迁移和侵袭。

Objective To investigate the effects of atorvastatin calcium on thyroid function, immune response and C-Jun N-terminal kinase/p38 mitogen-activated protein kinase (JNK/p38 MAPK) signaling pathway in rats with hypothyroidism. Methods A total of 30 healthy adult male SD rats were randomly divided into control group, hypothyroid group (PTU group) and atorvastatin calcium treatment group (ACT group), with 10 rats in each group. Rats in the PTU group and the ACT group were injected with PTU subcutaneously at the dorsum of the neck every day for 28 consecutive days; instead of PTU, rats in the control group were injected subcutaneously with 0.3 mL of saline. After 2 weeks of PTU treatment, rats in the ACT group were gavaged with 3 mL of atorvastatin calcium saline solution (containing 5 mg/kg of atorvastatin calcium), which was administered once daily; the control group was gavaged with an equal amount of saline in the same way. The body weight, food intake and water intake of rats were measured weekly. The histopathological changes of the thyroid gland were observed in histopathological sections of rats in each group. Enzyme-linked immunosorbent assay (ELISA) was performed to determine the levels of triiodothyronine (T3), thyroxine (T4), thyroid stimulating hormone (TSH), interferon γ (IFN-γ) and interleukin-4 (IL-4) in serum; quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was performed to detect the mRNA expression levels of IFN-γ, IL-10, Foxp3 and IL-4; western blot was performed to determine the levels of p-JNK/JNK and p-p38/p38 MAPK. Results Compared with control group, PTU-induced hypothyroidism rats showed a significant decrease in body mass and food and water consumption (P < 0.05). After 2 weeks of treatment with atorvastatin calcium, the body mass loss of PTU rats was inhibited, food and water consumption was improved, and the differences were statistically significant (P < 0.05). Atorvastatin calcium was able to significantly increase the serum T3 and T4 levels and decrease the serum TSH level in hypothyroid rats (P < 0.05). Atorvastatin calcium treatment was able to significantly alleviate histopathological changes such as follicular cell proliferation and related hypertrophy induced by PTU, and increase follicular size (P < 0.05). After treatment with atorvastatin calcium, the spleen mass of PTU rats increased significantly, and the expression of IFN-γ mRNA in hypothyroid rats decreased significantly, but the expression levels of IL-10 mRNA, Foxp3 mRNA and IL-4 mRNA increased significantly (P < 0.05). After treatment with atorvastatin calcium, the levels of p-JNK/JNK and p-p38/p38 MAPK in thyroid tissue of hypothyroid rats decreased significantly (P < 0.05). Conclusion Atorvastatin calcium treatment for hypothyroidism has the function of promoting the normalization of thyroid hormone imbalance, balancing Th1/Th2 cytokines, and inhibiting the activation of JNK/p38 MAPK signaling pathway.

Objective:To evaluate the ultrasound diagnostic value of portal vein complications after liver transplantation by monitoring changes in portal vein hemodynamic parameters using the color Doppler ultrasound technology and to determine its clinical significance.Methods:The clinical data of 99 patients who underwent liver transplantation at the Organ Transplantation Center of the Affiliated Hospital of Qingdao University from July 2015 to December 2018 were analyzed retrospectively. There were 81 males and 18 females, aged (51±9) years old. These patients were divided into the portal vein complication ( n=23) and the non-portal vein complication ( n=76) groups, based on whether portal vein complications had developed within 2 years after surgery. In addition, 30 healthy volunteers at the Affiliated Hospital of Qingdao University, including 16 males and 14 females, aged (40±14) years old were selected to form the control group. The patients’ morphology of liver was studied using color Doppler ultrasound at days 1, 7, 14, 30, 180, 365 and 730 after liver transplantation, and the maximum portal vein blood flow velocity and portal blood flow were recorded. Results:Compared with the control group, the maximum portal venous flow velocity and portal venous blood flow significantly increased on days 1, 7, 14, 30, and 180 after liver transplantation in the non-portal complication group (all P<0.05). With time, these changes showed a decreasing trend. By day 365 after surgery, the differences between the maximum portal venous flow velocity and the portal venous blood flow between the two groups became not significant ( P>0.05). Of the 23 patients in the portal vein complication group, 9 developed portal vein stenosis (PVS) and 14 portal vein embolism. The 9 patients with PVS had a maximum portal flow velocity of 63.8 (46.0, 78.6) cm/s at 1 month after surgery, and this flow velocity was significantly higher than that in the non-portal complication group [35.0(29.6, 41.8) cm/s, Z=-3.35, P<0.001]. The portal blood flow was 993 (887, 1168) ml/min in the 9 patients with portal vein stenosis at 1 month after surgery, and it was significantly higher than those in the non-portal complication group [811(682, 1 018) ml/min, Z=-2.37, P=0.020]. Conclusions:After liver transplantation, both the portal venous blood flow velocity and the blood flow were at high levels in the early postoperative period and they returned to normal levels with time. Ultrasound dynamic monitoring of portal venous blood flow changes was of clinical significance in diagnosing portal vein stenosis and portal vein embolism after liver transplantation.

Objective: To describe the MICM (morphology, immunology, cytogenetics and molecular biology) characteristics of a case of acute myelomonocytic leukemia M 4C . Methods: The medical history data of the case of M 4C admitted to our hospital was reviewed. The results of bone marrow cell morphology, cytochemical stains, bone marrow biopsy, immunophenotype, cytogenetics, molecular test and NGS (next-generation sequencing) of the case were analyzed. Results: The bone marrow smear showed markedly active proliferation of bone marrow cells in which the myelomonocytic cells accounted for 85.6%. Cytochemical stains showed peroxidase (POX) stain partially and weakly positive; specific esterase AS-DCE partially positive; non-specific esterase α-NBE partially positive and smothered by sodium fluoride; non-specific esterase AS-DAE partially positive and smothered by sodium fluoride. Bone marrow biopsy showed hyperproliferative cells and diffused hyperplasia of blasts. Immunophenotype analysis showed that the abnormal cell population was positive for CD11B, CD64, CD56, cMPO, CD33, CD41, CD61, CD38 and CD58, but negative for CD13, CD34, CD117, CD7, CD123, HLA-DR, CD10, CD19, CD20, CD2, CD14, CD235, CD15, CD303, CD304, CD25, cCD79a, cCD3, cCD22, CD1a and TDT. Cytogenetic analysis showed 47, XY, t(9;11) (p22;q23),+mar. The molecular test for leukemia showed MLLT3/KMT2A gene rearrangement. NGS showed NRAS and TET2 mutation. The case was finally diagnosed as AML (acute myelomonocytic leukemia) M 4C with t(9;11)(p22;q23), MLLT3-KMT2A. Conclusion Leukemia M 4C may show the characteristics of both granulocytes and monocytes with complex morphological features. The combined examination of MICM should be necessary for the diagnosis of M 4C with great significance.

Objective To evaluate the efficacy of volume therapy guided by stroke volume variabil-ity ( SVV) in the patients undergoing surgery for severe traumatic brain injury. Methods Thirty patients of both sexes with severe traumatic brain injury, aged 18-64 yr, of American Society of Anesthesiologists physical status Ⅲ, who were admitted to the hospital within 24 h after injury, with Glasgow Coma Scale ( GCS) score≤8, were divided into control group ( C group, n=15) and SVV group ( n=15) using a ran-dom number table method. In group C, conventional fluid administration was performed to maintain mean arterial pressure at 65-110 mmHg, central venous pressure at 5-12 cmH2 O and urine volume>1 ml·kg-1 ·h-1 . Fluid was given according to SVV, maintaining SVV≤13％ and mean arterial pressure at 65-110 mmHg in group SVV. Immediately after skin incision ( T0 ) , immediately after opening cerebral dura mater ( T1 ) , at 1 h after opening cerebral dura mater ( T2 ) , immediately after suturing cerebral dura mater ( T3 ) and at the end of operation ( T4 ) , blood samples were collected from the radial artery and inter-nal jugular venous bulb for blood gas analysis, the jugular venous oxygen partial pressure, jugular venous bulb oxygen saturation, blood lactate, arterial oxygen partial pressure, arterial oxygen saturation and Hbwere recorded, and the cerebral artery and arteriovenous blood O2 content difference and cerebral O2 extrac-tion rate were calculated. Blood samples were collected from the internal jugular venous bulb at T0-2 , T4 and 24 h after operation ( T5 ) for determination of S100β protein concentrations by enzyme-linked immunosor-bent assay. The intraoperative volume of fluid intake and output and consumption of vasoactive drugs were recorded. GCS scores were recorded immediately after admission to the operating room, and at 1, 3, 7 and 14 days after operation. The development of postoperative length of hospitalization and complications ( pul-monary infection and brain edema) was recorded. Glasgow Outcome Scale Score was used to assess the early postoperative quality of life. Results Compared with group C, the urine volume was significantly in-creased, the consumption of vasoactive drugs was reduced, jugular venous bulb oxygen saturation was in-creased at T2,3 , the cerebral O2 extraction rate was decreased at T2-4 , the serum S100β protein concentra-tion was decreased at T2 , and the GCS score was increased at day 3 after operation ( P<0. 05) , and no sig-nificant change was found in blood lactate, postoperative Glasgow Outcome Scale score or length of hospital-ization at each time point in group SVV ( P>0. 05) . Conclusion SVV-guided volume therapy can improve cerebral oxygen metabolism, ensure adequate tissue perfusion and reduce craniocerebral injury in the pa-tients undergoing surgery for severe traumatic brain injury.

Objective To evaluate the efficacy of phloroglucinol combined with dyclonine hydrochloride mucilage in preventing catheter-related bladder discomfort (CRBD)during recovery from anesthesia in patients under general anesthesia.Methods A total of 120 male patients scheduled for laparoscopic cholecystectomy under general anesthesia,aged 18-60 years,weighing 46-80 kg, ASA physical status I or II,were randomly divided into 3 groups (n=40 in each group):group of combination of phloroglucinol and dyclonine hydrochloride mucilage (group P),sufentanil group (group S)and control group (group C).After induction of general anesthesia,the patients in group P were tracheally incubated and then inj ected with 5 ml dyclonine hydrochloride mucilage per urethra.In the meantime,for patients of groups S and C,equal volume of normal saline was inj ected and paraffin oil was used to lubricate for urethral catheterization.The catheter was clamped and then reopened 30 min later.At 15 min before the end of surgery,80 mg Phloroglucinol,0.10 μg/kg sufentanil and an equal volume of normal saline were injected intravenously in group P,group S and group C,respec-tively.The catheter was removed when the patients were fully awake.The awakening time and extu-bation time were recorded.In addition,Riker sedation-agiation scale (SAS)score was documented at 5 min (T1),15 min (T2),30 min (T3),1 h (T4)and 2 h (T5)after extubation.The occurrence and severity of CRBD within 2 h after surgery,as well as occurrence of nausea and vomiting and respira- tory depression were recorded.Results Compared with group C,the SAS score at T1-T4and inci-dence and severity CRBD were decreased,whereas the emergence time and extubation time were pro-longed in group S.The SAS score at T1-T5,incidence and severity of CRBD were decreased (P<0.05),and no significant change was found in emergence time and extubation time in group P.Com-pared with group S,the SAS score at T1-T4was increased,whereas the SAS score at T5,incidence and severity of CRBD were decreased,and the emergence time and extubation time were shortened in group P (P<0.05).There was no significant difference in the incidence of nausea and vomiting,re-spiratory depression and extubation time among the three groups. Conclusion Dyclonine hydrochloride mucilage injected per urethra combined with phloroglucinolol injected intravenously at 1 5 min before the end of surgery can reduce the incidence and severity of CRBD during the recovery from anesthesia in the patients under general anesthesia.

Objective To explore the impact of neoadjuvant therapy on the number of harvested lymph nodes in D2 radical resection of the proximal locally advanced gastric cancer (GC).Methods The retrospective cohort study was conducted.The clinicopathological data of 319 patients with proximal locally advanced GC who were admitted to the Beijing Cancer Hospital from January 2013 to September 2016 were collected.Of 319 patients,200 underwent D2 radical resection of GC and didn't undergo neoadjuvant therapy who were divided into the surgery group,88 underwent neoadjuvant chemotherapy into the chemotherapy group,and 31 underwent neoadjuvant chemoradiotherapy into the chemoradiotherapy group.Observation indicators and evaluation criteria:comparison of postoperative pathological results among 3 groups,according to tumor staging guideline of American Joint Committee on Cancer (AJCC) (8th version) Measurement data with normal distribution were represented as x±s,comparisons among groups were analyzed using the ANOVA.Measurement data with skewed distribution were described as M (range),comparisons among groups were analyzed using the ANOVA,and pairwise comparisons were analyzed using nonparametric test.Comparisons of count data among groups were analyzed using the exact chisquare test,and pairwise comparisons were analyzed using the chi-square partition method.Results Comparison of postoperative pathological results among 3 groups:stage T0,Tla,Tlb,T2,T3,T4a and T4b of T staging were respectively detected in 0,2,10,24,99,58,7 patients in the surgery group and 5,1,2,11,41,26,2 patients in the chemotherapy group and 5,1,2,8,10,4,1 patients in the chemoradiotherapy group.Stage N0,N1,N2,N3a and N3b of N staging 56,41,34,47,22 patients in the surgery group and 29,17,27,10,5 patients in the chemotherapy group and 18,10,2,1,0 in the chemoradiotherapy group.Cases with and without lymphovascular invasion were respectively 124,76 in the surgery group and 43,45 in the chemotherapy group and 6,25 in the chemoradiotherapy group.Total number of harvested lymph nodes,number of lymph node metastases and rate of lymph node metastasis in the surgery,chemotherapy and chemoradiotherapy groups were respectively 31 (range,15-87),30 (range,15-62),21 (range,15-36) and 3 (range,0-39),2 (range,0-37),0 (range,0-7) and 9.2％ (range,0-91.3％),7.7％ (range,0-78.7％),0 (range,0-30.4％).There were statistically significant differences in the T staging,N staging,with and without lymphovascular invasion,total number of harvested lymph nodes,number of lymph node metastases and rate of lymph node metastasis among groups (x2 =35.799,32.489,21.076,Z =27.120,22.088,16.947,P ＜ 0.05).There were statistically significant differences in the above indicators between surgery group and chemoradiotherapy group (x2 =28.500,20.124,19.570,P＜0.05),and no statistically significant difference in the above indicators between surgery group and chemotherapy group (x2 =11.436,12.343,4.295,P＞ 0.05).There were statistically significant differences in the N staging,with and without lymphovascular invasion,total number of harvested lymph nodes,number of lymph node metastases and rate of lymph node metastasis between chemotherapy group and chemoradiotherapy group (x2 =14.027,8.313,P＜ 0.05),and no statistically significant difference in the T staging between chemotherapy group and chemoradiotherapy group (x2=11.742,P＞ 0.05).Conclusion Neoadjuvant chemoradiotherapy could reduce the total number of harvested lymph nodes and number of lymph node metastases after radical resection of proximal locally advanced GC.

Objective To evaluate the effect of low-dose norepinephrine (NE) combined with goal-directed fluid therapy (GDFT) on cerebral oxygen metabolism in patients undergoing intracranial tumor resection.Methods Forty patients of American Society of Anesthesiologists physical status Ⅱ or Ⅲ,aged ≥ 18 yr,scheduled for elective intracranial tumor resection,were divided into 2 groups (n =20 each) using a random number table method:GDFT group (group G) and low-dose NE combined with GDFT group (group N).Fluid was replaced according to stroke volume variation (SVV) under the guidance of Flotrac-Vigileo system in both groups.When SVV ≤13％,fluid was replaced at 1-2 ml · kg-1 · h-1.When SVV＞ 13％ for more than 5 min,fluid replacement was enhanced to reduce it below 13％.In group N,NE was infused continuously via the central vein at 0.01-0.03 μg· kg-1 · min-1 after anesthesia induction,and mean arterial pressure (MAP) was maintained ≥ 65 mmHg.After anesthesia induction (T1),when the dura of brain was opened (T2),at 1 h after opening the dura (T3) and at the end of surgery (T4),the heart rate and MAP were recorded,and blood samples were collected from the internal jugular venous bulb and radial artery for blood gas analysis.The fluid input and output were recorded.Arterial oxygen content,jugular bulb venous oxygen content,arteriovenous oxygen content difference,cerebral oxygen extraction rate,cerebral lactic acid production rate and ratio of cerebral blood flowto cerebral oxygen metabolic rate were calculated.Results Compared with group G,MAP at T4 and cerebral oxygen extraction rate at T3,4 were significantly increased,the total volume of fluid and volume of crystalloid solution were decreased (P＜0.05),and no significant change wasfound in arterial oxygen content,jugular bulb venous oxygen content,arteriovenous oxygen content difference,ratio of cerebral blood flow to cerebral oxygen metabolic rate or cerebral lactic acid production rate in group N (P＞0.05).Conclusion Low-dose NE combined with GDFT can reduce the intraoperative volume of fluid infused and improve cerebral oxygen supply when applied to the patients undergoing intracranial tumor resection.

Objective:To establish an LC-MS/MS method for the content determination of triptolide in Shenshe ointment.Methods:The chromatographic separation was performed on an Agilent TC-C18 (250 mm×4.6 mm,5 μm)column with the mobile phase consisting of methanol-0.1% formic acid solution (73∶27,v/v).The flow rate was 0.5 ml·min-1 and the column temperature was maintained at 35℃.An electrospray ionization (ESI) source was applied with multiple reaction monitoring (MRM) combined with a positive mode.Metronidazole was used as the internal standard.The mass transitions were 361.4→43.2 for triptolide and 172.1→128.0 for metronidazole,respectively.Results:The linear range was 10-500 ng·ml-1 with good correlation coefficient (r=0.999 9).The limit of quantification (LOQ) was 9.5 ng·ml-1.The intra-and inter-day RSDs of peak areas were all less than 3% and the average recovery was 96.87%(RSD=2.79,n=6).Conclusion:The established LC-MS/MS method is simple,efficient,sensitive and accurate in the quality control of Shenshe ointment.