Objective To study the role of glutamate receptor subtypes in nucleus tractus solitarius(NTS)in cardiac-somatic motor reflex (CMR)induced by intrapericardial administration of capsaicin,and to clarify the modulation mechanism of NTS to cardiac nociceptoion.Methods 60 SD rats were randomly divided into ibotenic (IBO)group, glutamate group, MK-801 group, MCGP group, MK-801 + MCPG group and DNQX group. The NTS microinjected with 130 mmol·L-1 IBO 100 nL,100,200,500 mmol·L-1 L-glutamate 100 nL,NMDA receptor antagonist 40 and 60 mmol · L-1 MK-801 100 nL, metabotropic glutamate receptors antagonist 25 and 50 mmol·L-1 MCPG 100 nL,25 mmol· L-1 MCPG 50 nL plus 40 mmol· L-1 MK-801 50 nL,non-NMDA receptor antagonist 20 and 50 mmol·L-1 DNQX 100 nL,respectively.The changes of CMR of the rats in various groups were observed.Results Compared with control group,the CMR of the rats in IBO group was decreased (P<0.05);the CMR was increased as the concentration increased in glutamate group(P<0.05);the CMR in MK-801 and MCPG groups were decreased (P<0.05);the CMR in MK-801+MCPG group was decreased (P<0.05);the CMR in DXQX group had no changes (P>0.05).Conclusion NTS play an facilictory role in cardiac nociception,and the NMDA receptors and mGluRs receptors mediate this facilitory modulation.

Objective To explore the role of metabotropic glutamate receptors (mGluRs)group Ⅲ and its subtypes mGluR7 and mGluR8 in nucleus tractus solitarius(NTS)in cardiac-somatic motor reflex (CMR),and to clarify the modulation role of mGluR Ⅲ and its subtypes in NTS in cardiac nociceptoion.Methods 40 SD rats were randomly divided into L-AP4 group,microinjection of mGluRs Ⅲ agonist L-AP4 0.1,1.0,10.0 or 20.0 nmol in NTS;AMN082 group,microinjection of mGluR7 agonist AMN082 1,2 or 4 nmol;DCPG group,microinjection of mGluR8 agonist DCPG 4, 6 or 8 nmol;MSOP group, microinjection of mGluR Ⅲ antagonist MSOP 20 or 100 nmol,20 nmol MSOP+410 nmol L-AP,20 nmol MSOP+2 nmol AMN082,20 nmol MSOP+6 nmol DCPG. The changes of CMR of the rats in various groups were observed.Results Compared with control,the CMR in L-AP4 and AMN082 groups was decreased (P<0.05);the CMR in DCPG group was increased (P<0.05);the CMR in MSOP group after injection of 20 nmol MSOP had no change (P>0.05);the CMR in MSOP group after injection of 100 nmol MSOP was increased (P<0.05);the CMR in MSOP group after injection of 20 nmol MSOP followed L-AP4 or AMNO82 had no change (P>0.05).Conclusion The group Ⅲ mGluRs in the NTS play an inhibitory role in cardiac nociception, and mGluR7 has anti-nociceptive effects while mGluR8 has pro-nociceptive effects.

Objective:To express the GPI-anchored CD55 and recombinant transmembrane form CD55 molecules on mouse fibroblasts cell line NIH3T3, and compare their inhibitory function of complement lysis.Methods:In previous study,had constructed the transmembrane form CD55 (CD55-TM) cDNA by linking the extracellular portion of CD55 to the transmembrane and cytoplasmic domains of MCP, and then subcloned into retroviral vector pLXSN. In this experiment,had transfected recombinant CD55-TM and GPI anchored CD55 into PA317 packaging cell to generate stable virus-producing cell lines. And then, mouse fibroblasts cell line NIH3T3 was infected with the virus containing CD55, recombinant CD55-TM or pLXSN alone. The expression of these molecules on NIH3T3 cells was detected by FACS analysis. Complement killing assay was carried out using MTT colorimetric method.Results:FACS analysis showed that both CD55 and its TM version were expressed stably on NIH3T3 cells. Both kinds of CD55 molecules can efficiently protect the NIH3T3 cells from complement mediated lysis and there is no significant difference between them. Conclusion:These data showed that both GPI-anchored CD55 and its TM version can normally expressed on NIH3T3 cells and both can protect the NIH3T3 cells from human complement mediated lysis. These results confirmed the feasibility of TM CD55 based gene therapy could be used for PNH, and also provided an excellent model for the study of signal transduction mechanisms mediated by GPI-anchored protein.

Objective To study the clinical features,diagnosis and management of small cell carcinoma of the bladder (SCCB).Method The clinical data of 18 cases of patients with small cell carcinoma of the bladder were analyzed retrospectively and the literature were reviewed.There were 16 males and 2 females,ages 54 to 81 years (median age,61 years).Clinical manifestations included gross hematuria in 11 cases,urgency in 2 cases,dysuria in 2 cases and postoperative review after TURBT of bladder urothelial carcinoma in 3 cases.The median tumor size was 3.35cm (ranged,1.0 to 6.0 cm).2 cases underwent TURBT and intravesical chemotherapy regularly were followed after surgery.3 cases underwent partial cystectomy,intravenous chemotherapy combined with radiotherapy was followed in one case,the other 2 cases refused the following therapy.13 cases underwent radical cystectomy,intravenous chemotherapy was followed in 2 cases,pelvic radiotherapy was followed in 2 csaes and intravenous chemotherapy combined with radiotherapy was followed in 2 cases,the other 7 cases refused the following therapy.Results 11 cases were pure SCCB,7 cases were mixed SCCB,all with urothelial carcinoma.T1N0M0 in 3 cases,T2N0.1M0 in 4 cases,and T3N0-2M0 in 11 cases.The duration of follow-up was from 5 to 35 months after surgery.9 cases died of tumor metastasis,9 cases are still alive,except 1 case with lymph node metastasis,the other 7 cases are free of tumor recurrence or metastasis.Conclusions SCCB is rare,with high malignant degree and poor prognosis.The prognosis of the pure SCCB may be worse than the mixed SCCB.The diagnosis depends on pathology examination.Radical cystectomy is the main treatment method,the strategy of bladder-preserving may be an attempt for proper SCCB patients.Adjuvant therapy plus surgery may be better.

Mandibular symphysis fracture combined with anteromedial dislocated condyle fracture is commom in clinical,but mandibular symphysis fracture associated with superolateral dislocation of the mandibular condyle is rare,which is often misdiagnosed or completely over-looked.Malpractice can lead to ankylosis and other sequelae.This article reviews 10 patients with mandibular symphysis fracture associated with superolateral dislocation of the mandibular condyle,discusses the causative mechanism,diagnostic features and clinical management according to literature data.

Objective:To compare the effect of fluorescence staining and periodic acid-Schiff staining in the diagnosis of fungal keratitis (FK).Methods:A total of 147 corneal specimens from 147 FK patients treated in Eye Hospital of Shandong First Medical University from January 2017 to May 2019 with positive corneal scraping or fungal culture were collected.Among them, there were 84 cases with penetrating keratoplasty (PKP), 42 cases with lamellar keratoplasty (LKP) and 21 cases with lesion resection.Another 11 cases with herpes simplex virus keratitis served as negative control.The corneal tissue specimens were performed with fungal fluorescence staining and periodic acid-Schiff staining, respectively.The stained sections were placed under fluorescence microscope and optical microscope to observe fungal hyphae or spores, respectively.The positive rates of the two staining methods were compared, and the positive cases of the diagnosis of FK in corneal tissue samples obtained by different surgical methods and corneal infection caused by different strains of the two staining methods were compared.Written informed consent was obtained from each patient.The study protocol adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Shandong Eye Hospital (No.SDSYKYY-2016012).Results:The positive rate of periodic acid-Schiff staining and fungal fluorescence staining was 60.5% (89/147) and 79.6% (117/147), respectively.The positive rate of fluorescence staining in the diagnosis of fungal keratitis was significantly higher than that of periodic acid-Schiff staining ( χ2=28.00, P<0.01), and both the specificity of the two staining methods was 100%.The positive rate of specimens obtained by PKP with fluorescent staining was 85.7% (72/84), and the positive rate with periodic acid-Schiff staining was 65.5% (55/84), and the difference was statistically significant ( χ2=17.00, P<0.01). The positive rate of specimens obtained by LKP with fluorescent staining was 71.4% (30/42), and the positive rate with periodic acid-Schiff staining was 52.4% (22/42), and the difference was statistically significant ( χ2=8.00, P<0.01). The positive rate of resected foci specimens with fluorescent staining was 71.4% (15/21), and the positive rate with periodic acid-Schiff staining was 57.1% (12/21), and the difference was not statistically significant ( χ2=1.30, P=0.25). The positive cases of two kinds of staining were different among different fungal strains.Among them, the positive cases of Fusarium solani complex, Pythium insidiosum, Aspergillus fumigatus complex, Candida guilliermondii, Trichoderma and Nigrograna mackinnonii with fluorescence staining were 19, 5, 5, 1, 1 and 1, and the positive cases of periodic acid-Schiff staining were 11, 0, 3, 0, 0 and 0, respectively.The staining results of the 11 negative controls were negative. Conclusions:Fluorescence staining is more sensitive than periodic acid-Schiff staining in the detection of fungal components in paraffin-embedded corneal tissues, and it can significantly improve the fungal detection rates.

Objective To study the value of CT perfusion imaging(CTPI)on brain hemodynamic of the aged with cerebral infarction. Methods The 48 patients who were doubted with cerebral infarction underwent 16-slice CT plain scanning and CTPI within 24 hours of onset. The cerebral blood flow(CBF), mean transit time(MTT)and time to peak(TTP)of contrast-medium in region of interest(FOV)were used as brain hemodynamic parameters in comparation with contralateral regions. All cases were followed up with MRI after 3-10 days. Results Ischemia lesion was found on CT plain scanning in 40.9% of patients, while 93.2% of patients showed abnormal perfusion on CTPI. The sensibility of CTPI in identifying ischemia area was 93.2%, and the specificity was 100%. CBF in research area was significantly reduced, MTT and TTP were remarkably increased in contrast to counterparts(P＜0.01). Conclusions CT perfusion imaging can sensitively reveal the hemodynamic condition of cerebral ischemia, which could provide the important information for early diagnosis and treatment of the elderly with brain infarction.

AIM:To investigate the application value of fluorescent staining technique in the detection of amoebic pathogens in corneal tissue biopsy, and to apply fluorescent staining technique in the histopathological diagnosis of Acanthamoeba keratitis(AK), comparing the results with those of hemotoxyiln-eosin staining(HE staining)and periodic acid-schiff staining(PAS staining), and analyzing the sensitivity and specificity of these three staining methods.

METHODS:Specimens of infected corneal tissue were collected from 74 cases(75 eyes), and then they were divided into an AK group and a non-Acanthamoeba keratitis(NAK)group based on the results of corneal scraping, culture and histopathological diagnosis. The tissues of consecutive sections were stained with HE staining, PAS staining and fluorescence respectively, and the sensitivity and specificity of the three staining methods for the diagnosis of AK were analyzed. Area under the curve(AUC)was calculated using the receiver operating characteristic(ROC)curve. Further analysis was performed to count the number of Acanthamoeba pathogens found by the three staining methods under the same magnification field of view at the same site, and to clarify the diagnostic value of fluorescent staining technique for AK.

RESULTS: The sensitivity of HE staining was 69%(27/39)with a specificity of 92%; the sensitivity of PAS staining was 62%(24/39)with a specificity of 97%, and the sensitivity of fluorescent staining was 95%(37/39)with a specificity of 97%. There were differences in the sensitivity of the three staining methods for the diagnosis of AK(χ2=19.857, P<0.001), and pairwise comparison revealed that the differences between HE staining and fluorescent staining, PAS staining and fluorescent staining for the diagnosis of AK were statistically significant(P=0.003,<0.001), while the difference in sensitivity between HE staining and PAS staining for the diagnosis of AK was not statistically significant(P=0.978). The maximum AUC was 0.960 for fluorescence staining, followed by 0.804 for HE staining and 0.794 for PAS staining, respectively. The median number of amoeba cysts detected by HE staining, PAS staining and fluorescent staining at the same site under the same magnification field of view was 4(0, 11), 2(0, 9)and 12(3, 33), respectively(χ2=56.561, P<0.001). Pairwise comparison revealed that the differences in the number of amoeba cysts found by HE staining and fluorescence staining, PAS staining and fluorescence staining were statistically significant(P<0.001), while the difference in the number of amoeba cysts found by HE staining and PAS staining was not statistically significant(P=0.210). Fluorescently stained histopathological sections make it easier to identify amoebic pathogens.

CONCLUSION:Fluorescent staining technique is more sensitive to histopathological diagnosis of AK than HE staining and PAS staining, which can significantly improve the positive rate of detection of amoebic pathogens.

Objective To investigate the expression of DNA-methyltransferases 3B(DNMT3B)gene in human pancreatic carcinoma and to evaluate its relationship with elinicopathologic parameters.Methods 42 samples of pancreatic carcinoma tissues and 42 para-carcinoma tissues and 10 normal pancreatic tissues were collected and the expression of DNMT3B mRNA and protein Was detected by real.time PCR and immunohistochemistry techniques.Results The expression of DNMT3B mRNA(RQ level)in human pancreatic carcinoma tissues and para-carcinoma tissues,normal pancreatic tissues was 9.4±5.9,1.02±0.71 and 0,respectively,and the difference was statistically significant(P<0.05).The rate of expression of DNMT3B protein in human pancreatic carcinoma tissues,para-carcinoma tissues and normal pancreatic tissues were 83.3%,14.3%and 10%,respectively,and the difference wag also statistically significant(P<0.01).The expression of DNMT3B mRNA correlated significantly with clinical staging,differentiation degree of the tumor and lymph node metastasis(P<0.01 or P<0.05).The expression of DNMT3B protein correlated significantly with the location ofthe tumor and lymph node metastasis(P<0.01 or P<0.05).The expression of DNMT3B mRNA and protein Was not assecimed with age,sex,neural invasion,tumor size,sernm CEA and CA19-9.Conclusions Highly expressed DNMT3B mRNA and protein may indicate the lymph node metastasis and poor prognosis in human pancreatic carcinoma.

Objective To investigate the feasibility of induction of experimental chronic pancreatitis rat model with L-arginine.Methods Animals were randomly divided into control group,arginine 12 h group,arginine 24 h group and arginine 7 d group with 10 rats in each group.L-arginine solution was intraperitoneally injected twice with an interval of 1 h.Serum amylase and glucose levels at corresponding time points were detected and histopathological scores of pancreas were evaluated.Collagen in pancreas was stained with Van Gieson method.Results Serum amylase levels were (1 634±890 ) U/L,( 3 872±2 676 ) U/L,( 3 307±2 197)U/L and (1 561±304) U/L in control group,arginine 12 h group,arglnine 24 h group and arginine 7 d group,respectively.The serum amylase level in arginine 7 d group was significantly lower than those in arginine 12 h group and arginine 24 h group (P ＜ 0.05 ).There was no significant difference in serum glucose level among all the groups.Histopathological scores were 0.8±0.4,5.1±2.6,6.5±2.2 and 4.5±1.6,respectively.The histopathological score of arginine 7 d group was significantly lower than those in arginine 24 h group (P ＜ 0.05 ).Obvious collagen could be found in pancreatic parenchyma in arginine 7 d group,while little collagen was found in pancreatic tissue in control,arginine 12 h and arginine 24 h groups.Conclusions Injection of L-arglnine induced fibrosis in pancreatic parenchyma and proliferation of tubular complex 7 days later,and it could be used for chronic pancreatitis model induction.