[Expression of stem cell markers CD44 and Lgr5 in colorectal cancer and its relationship with lymph node and liver metastasis].

Objective: To investigate the relationship between the expression of stem cell markers CD44 and Lgr5 and the clinicopathological features, lymph node and liver metastasis, and to analyse the value of both in evaluating the prognosis of colorectal cancer. Methods: A total of 90 cases of colorectal cancer, 20 cases of adjacent tissues, 30 cases of lymph node metastases, 28 cases of cancer nodules and 30 cases of liver metastasis of colorectal cancer were collected from Department of Pathology of the Second Hospital of Tianjin Medical University from January 2011 to June 2016. The expression of CD44 and Lgr5 protein was detected by immunohistochemistry, and the relationship between them and clinical pathological data, lymph node metastasis and liver metastasis were analyzed. Results: The positive expression rates of CD44 and Lgr5 in colorectal cancer tissues were 68.9% and 62.2%, which were significantly higher than 30.0% and 15.0% of adjacent tissues(P=0.001 and P<0.001). The expression of CD44 and Lgr5 in colorectal cancer was associated with histological differentiation, depth of tumor invasion, lymph node metastasis, soft tissue nodules, hepatic metastasis, and TNM staging(P=0.021, 0.032, 0.030, 0.011, 0.015, 0.007 and P=0.011, 0.027, 0.017, 0.008, 0.011, 0.021). The positive rates of CD44 and Lgr5 expression in lymph node metastases, cancer nodules and liver metastases (93.3%, 89.3%, 90.0%, and 83.3%, 89.3%, 86.7%) were higher than those in the primary lesions(68.9% and 62.2%)(P=0.007, 0.032, 0.022 and P=0.033, 0.007, 0.013). There is a positive correlation between the expression of CD44 and Lgr5 in colorectal cancer(r=0.615, P<0.001), and both positive expression of CD44 and Lgr5 is associated with histological differentiation, lymph node metastasis and cancer nodules, liver metastasis and TNM staging(P=0.005, 0.003, 0.003, 0.026, 0.014). The Kaplan-Meier survival curve showed that the survival rate of positive expression of CD44 and both positive expression of CD44 and Lgr5 in patients with colorectal cancer was low(P=0.030 and 0.001). Log-rank univariate survival analysis showed that histological differentiation, depth of invasion, lymph node metastasis, liver metastasis, and clinical stage were the influencing factors of postoperative survival time(P=0.035, 0.035, 0.018, 0.016, 0.004). Cox proportional hazards regression model multivariate analysis showed that both positive expression of CD44 and Lgr5, lymph node metastasis, liver metastasis, and clinical stage were independent factors affecting the prognosis of patients with colorectal cancer(P=0.004, 0.044, 0.031, 0.008). Conclusions: The expression of CD44 and Lgr5 is related to the malignant biological behavior of colorectal cancer, and they are important factors in promoting local and distant metastases. Both positive expression of them have a hint of bad prognosis.

Quantitative trait locus analysis of grape weight and soluble solid content.

A grapevine hybrid population was derived from a crossing of the early-maturing female parent cultivar '87-1' and the late-maturing male parent cultivar '9-22'. A total of 149 plants were selected from the hybrid population as the mapping population, and after sequence-related amplified polymorphism and simple-sequence repeat marker analysis were conducted we constructed molecular genetic maps of the parents. The molecular linkage map of '87-1' had 19 linkage groups that contained 188 markers, with an average interval of 5.7 cM and a total distance of 1074.5 cM; the '9-22' map had 19 linkage groups that contained 175 markers, with an average interval of 7.8 cM and a total distance of 1100.2 cM. The molecular linkage map of both parents had 19 linkage groups that contained 251 markers, with an average interval of 5.0 cM and a total distance of 1264.2 cM. We used the interval mapping method to conduct a quantitative trait locus (QTL) analysis of grape weight and soluble solid content of the mapping population. Six QTLs were related to grape weight, and the average contribution to the phenotypic variance was between 11.3 and 33.0%. Seven QTLs were related to soluble solid content, and the average contribution to the phenotypic variance was between 15.7 and 55.8%.

[Incidence and prognosis of olfactory and gustatory dysfunctions related to infection of SARS-CoV-2 Omicron strain: a national multi-center survey of 35 566 population].

Objective: This cross-sectional investigation aimed to determine the incidence, clinical characteristics, prognosis, and related risk factors of olfactory and gustatory dysfunctions related to infection with the SARS-CoV-2 Omicron strain in mainland China. Methods: Data of patients with SARS-CoV-2 from December 28, 2022, to February 21, 2023, were collected through online and offline questionnaires from 45 tertiary hospitals and one center for disease control and prevention in mainland China. The questionnaire included demographic information, previous health history, smoking and alcohol drinking, SARS-CoV-2 vaccination, olfactory and gustatory function before and after infection, other symptoms after infection, as well as the duration and improvement of olfactory and gustatory dysfunction. The self-reported olfactory and gustatory functions of patients were evaluated using the Olfactory VAS scale and Gustatory VAS scale. Results: A total of 35 566 valid questionnaires were obtained, revealing a high incidence of olfactory and taste dysfunctions related to infection with the SARS-CoV-2 Omicron strain (67.75%). Females(χ2=367.013, P<0.001) and young people(χ2=120.210, P<0.001) were more likely to develop these dysfunctions. Gender(OR=1.564, 95%CI: 1.487-1.645), SARS-CoV-2 vaccination status (OR=1.334, 95%CI: 1.164-1.530), oral health status (OR=0.881, 95%CI: 0.839-0.926), smoking history (OR=1.152, 95%CI=1.080-1.229), and drinking history (OR=0.854, 95%CI: 0.785-0.928) were correlated with the occurrence of olfactory and taste dysfunctions related to SARS-CoV-2(above P<0.001). 44.62% (4 391/9 840) of the patients who had not recovered their sense of smell and taste also suffered from nasal congestion, runny nose, and 32.62% (3 210/9 840) suffered from dry mouth and sore throat. The improvement of olfactory and taste functions was correlated with the persistence of accompanying symptoms(χ2=10.873, P=0.001). The average score of olfactory and taste VAS scale was 8.41 and 8.51 respectively before SARS-CoV-2 infection, but decreased to3.69 and 4.29 respectively after SARS-CoV-2 infection, and recovered to 5.83and 6.55 respectively at the time of the survey. The median duration of olfactory and gustatory dysfunctions was 15 days and 12 days, respectively, with 0.5% (121/24 096) of patients experiencing these dysfunctions for more than 28 days. The overall self-reported improvement rate of smell and taste dysfunctions was 59.16% (14 256/24 096). Gender(OR=0.893, 95%CI: 0.839-0.951), SARS-CoV-2 vaccination status (OR=1.334, 95%CI: 1.164-1.530), history of head and facial trauma(OR=1.180, 95%CI: 1.036-1.344, P=0.013), nose (OR=1.104, 95%CI: 1.042-1.171, P=0.001) and oral (OR=1.162, 95%CI: 1.096-1.233) health status, smoking history(OR=0.765, 95%CI: 0.709-0.825), and the persistence of accompanying symptoms (OR=0.359, 95%CI: 0.332-0.388) were correlated with the recovery of olfactory and taste dysfunctions related to SARS-CoV-2 (above P<0.001 except for the indicated values). Conclusion: The incidence of olfactory and taste dysfunctions related to infection with the SARS-CoV-2 Omicron strain is high in mainland China, with females and young people more likely to develop these dysfunctions. Active and effective intervention measures may be required for cases that persist for a long time. The recovery of olfactory and taste functions is influenced by several factors, including gender, SARS-CoV-2 vaccination status, history of head and facial trauma, nasal and oral health status, smoking history, and persistence of accompanying symptoms.

[Skull base aneurysms: a retrospective review of fifteen cases focusing on the involvement of internal carotid artery].

Objective: To explore the diagnosis and clinical features of internal carotid artery aneurysm in the skull base. Methods: The data of 15 patients with internal carotid aneurysms in the skull base diagnosed and treated by digital subtraction angiography (DSA) or CT angiography (CTA) in the Provincial Hospital Affiliated to Shandong First Medical University from 1995 to 2017 were collected and analyzed. Among the 15 patients, 12 were males, and 3 were females, aging from 17 to 67 years old, with a median age of 44 years. Thirteen patients were diagnosed by DSA; the other two patients were diagnosed by CTA. Thirteen patients were diagnosed with pseudoaneurysm with the first symptom of epistaxis, in which eight patients underwent head trauma and 5 underwent radiotherapy of skull base tumor. The other two patients were diagnosed with true aneurysm presented headache and cranial nerve disorder. All patients were followed up for 2 to 12 years after treatment to see whether they were cured and survived. Results: Among the eight patients with a history of trauma, five patients were cured by embolization, two patients without embolization died of massive epistaxis, one patient died of progressive cerebral infarction after embolization. Among the five patients with radiotherapy of skull base tumor, one patient died of cerebral infarction after embolization, two patients died out of the hospital due to the recurrence of the primary tumor and intracranial invasion, one patient recovered well after embolization and surgical operation, one patient gave up treatment and died of massive hemorrhage out of hospital. In the other two patients with symptom of headache, one received embolization treatment outside the hospital after receiving mistake operation, and another one gave up treatment and died due to personal reasons. In total, four patients died in hospital, four died out of the hospital, and seven patients survived. Conclusions: Internal carotid artery aneurysm is a high-risk disease of anterior and middle skull base. For patients with epistaxis with a history of trauma and radiotherapy or patients with headaches and cranial nerve disorders, the possibility of the internal carotid artery aneurysm should be considered, in which DSA or CTA examination is essentially required for ensured diagnosis and disease evaluation.. The correct diagnosis and treatment by the otolaryngologist are crucial to the prognosis of the patient.

[Resection and free flap repair of recurrent maxillofacial and paranasal sinus skull base malignant tumor: report of 9 cases].

Objective: To investigate the treatment method and effect of surgical resection and free flap repair of recurrent malignant tumors of maxillofacial paranasal sinus and skull base. Methods: The clinical data of 9 patients with recurrent maxillofacial paranasal sinus and skull base malignant tumors who underwent surgical resection and free flap repair in the Department of Otorhinolaryngology Head and Neck Surgery of Shandong Provincial Hospital from August 2009 to May 2019 were analyzed retrospectively, including 5 males and 4 females, aged 32-69 years. There were 4 cases of squamous cell carcinoma, 3 cases of adenoid cystic carcinoma, 1 case of myoepithelial carcinoma and 1 case of malignant fibrous histiocytoma. All patients were repaired with anterolateral femoral flap. The treatment process and postoperative effect were analyzed by descriptive statistics. Results: All patients were followed up from 16 months to 6 years. There were 6 cases of recurrence after operation, including 1 case of local recurrence and 5 cases of skull base and craniocerebral recurrence. The median tumor free survival time of recurrent patients was 35 months. Conclusion: Surgical resection and free flap repair can effectively prolong the life of patients with recurrent maxillofacial paranasal sinus and skull base malignant tumors.

[Granulomatosis with polyangiitis with nasal symptoms as the first clinical manifestation: clinical analysis of 18 cases].

Objective: To analyze and summarize the diagnosis, treatment and prognosis of granulomatosis with polyangiitis (GPA) with nasal symptoms as the first clinical manifestation. Methods: The data of 18 patients of GPA with nasal mucosal symptoms as the first clinical manifestation from the Department of Otorhinolaryngology Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University between 2005 and 2019 was collected, including 8 males and 10 females, aged from 5 to 68 years. Nasal endoscopy, imaging examination, laboratory examination, immunological and histopathological examination of nasal mucosa were completed. All patients were treated with glucocorticoid combined with cyclophosphamide and were followed up for 2 to 15 years. Descriptive statistical method was used for analysis. Results: All the 18 patients had the nasal mucosal symptoms as the first clinical manifestation, including nasal obstruction, running nose and epistaxis. Nasal endoscopy showed swelling, erosion, scab and bleeding of nasal mucosa, and 6 cases had nasal septal perforation. Nasal sinus CT scan showed high density shadow of sinus, as well as hyperostosis and osteosclerosis. CT imaging features of pulmonary showed nodular lesion or patchy infiltration in 12 patients and cavitation was found in 6 cases. Laboratory results showed that 13 cases were positive for anti-neutrophil cytoplasmic antibodies (ANCA), and 5 cases were negative. During follow-up period, thirteen patients were symptomatic controlled and survived; two patients died of disease progression; one patient gave up treatment and died; two patients were lost to follow-up. Conclusions: Nasal symptoms are the first clinical manifestation of GPA. Early diagnosis and early treatment with glucocorticoid combined with cyclophosphamide can effectively improve the survival rate.

[Reconstruction of orbital floor defect with preserved orbital contents after advanced maxillary sinus cancer resection].

Objective: To analyze the method and effect of reconstruction of the orbital floor defect with preserved orbital contents after advanced maxillary sinus cancer resection. Methods: Thirty-three patients of T3/T4 stage maxillary sinus cancer with orbital invasion who were treated in Shandong Provincial Hospital from January 2010 to October 2016 were retrospectively analyzed, including 20 males and 13 females, aged from 12 to 80 years old. Patients were treated with appropriate surgical methods according to their clinical manifestation, imaging finding, invasion range, preoperative and intraoperative pathology. Surgical approaches such as maxillectomy, expanded maxillectomy and superstructure maxillectomy were used for patients with Medpor orbit floor repairment. Patients were followed up regularly after surgery, and the surgical efficacy was discussed by descriptive statistical method. Results: Of those 33 cases, 19 were squamous cell carcinoma, 8 were adenoid cystic carcinoma, 5 were inverted papilloma malignant transformation, and 1 was myoepithelial carcinoma in our study. After tumor resection and orbital floor repair, the orbital contents and eyeball function of all patients remained intact. During follow-up time ranged from 3 to 8 years, a total of 6 patients relapsed at 3 years and died of ineffective treatment. The survival rate was 81.8% (27/33). Conclusion: Tumor resection and Medpor orbital floor repair in maxillary sinus cancer patients with orbital invasion can preserve the function of the eyeball well, greatly improve the quality of patient's life.

Sperm mediated human coagulation factor VIII gene transfer and expression in transgenic mice.

PRINCIPLES: B-domain deleted human coagulation factor VIII cDNA (BDD-hFVIIIcDNA) transgenic mice were produced by using sperm mediated gene transfer (SMGT). The transcription and expression of human FVIII in transgenic mice were also investigated. METHODS: Sperm were isolated from caudae epididymides of male C57BL/6 mice and transfected with linearized RC/RSV-BDDhFVIIIcDNA plasmid, and subsequently used to fertilize female mice via artificial insemination in vivo. After birth, F0 progeny were identified by PCR and Southern blotting for BDD-hFVIIIcDNA transgenic mice. F1 progeny were subsequently derived from a male transgenic F0 mouse and a normal C57BL/6 female mouse. The F1 progeny were then identified as BDD-hFVIIIcDNA transgenic mice by Southern blotting. The transcription and expression of BDDhFVIIIcDNA in transgenic mice were determined by northern blotting, western blotting and immunohistochemical staining. Blood was also collected from both F0 and F1 progeny to detect hFVIII:Ag and anti-hFVIII inhibitors. RESULTS: A total of 9 F0 and 8 F1 progeny were delivered, in which 3 F0 and 2 F1 progeny were identified to have BDD-hFVIIIcDNA. The transcription and expression of BDD-hFVIIIcDNA were found to exist in the liver and kidneys of all transgenic mice. HFVIII:Ag in plasma of the transgenic F0 progeny was 31.95 ng/ml, 23.52 ng/ml and 26.36 ng/ml respectively, whilst the F1 transgenic mice showed results of 18.82 ng/ml and 12.16 ng/ml. Anti-hFVIII inhibitors were negative in both F0 and F1 progeny. CONCLUSIONS: Human FVIII gene transgenic mice can be produced by the SMGT technique and express human FVIII protein in their bodies.

[Clinical analyses of the diagnosis and treatment of invasive fungal rhinosinusitis: report of 14 cases].

OBJECTIVE: Through the retrospective analysis of the clinical data in 14 cases of invasive fungal rhinosinusitis (IFRS), the clinical characteristics, diagnosis and treatment of this disease were evaluated. METHODS: Fourteen clinically confirmed cases of IFRS since January 2008 to October 2015 were evaluated.collected, the clinical features, diagnosis, treatment and prognosis were analyzed to obtain a more comprehensive understanding for clinical reference. Fourteen patients were confirmed by pathological examination as IFRS, including 9 cases of aspergillus, 4 cases of mucor, and 1 case of rhinocerebral zygomycosis; including 5 cases of acute IFRS, 9 cases of chronic IFRS. All patients were treated with endoscopic surgery and intravenous antifungal therapy. RESULTS: Nine cases of chronic IFRS (including 1 case of mucor, 7 cases of aspergillus and 1 case of rhinocerebral zygomycosis) were cured, but the vision loss, diplopia or blindness, hard palate perforation remained. Five cases of acute IFRS included 3 cases of mucor and 2 cases of aspergillus. Among the 3 cases of mucor, 2 cases were died and 1 case was cured. Among the 2 cases of aspergillus, 1 patient was cured and the other patient died of electrolyte disorder after discharge from hospital. CONCLUSIONS: Patients with IFRS usually have diabetes. After the active surgical cleaning of lesion tissue and the systematic antifungal treatment with adequate dosage, these patients would have a better result. IFRS caused by mucor is ofen dangerous.

[Effect of agonists of opioid receptor on coronary arterial strips in porcine].

Bioassay of spiral strips was used to study the effect of agonists of opioid receptor on coronary artery strips of porcine. It was found that selective agonist of kappa (U-50488H), mu (ohmefentanyl OMF) and delta (morphine and DADLE) receptor and wide spectrum agonist of opioid receptor etorphine all produced the contraction of strips dose-dependently. The order of the effect was U-50488H > morphine > etorphine > OMF, DADLE. Naloxone did not antagonize but increase the effect, or even produce contraction of the strips by itself. Haloperidol inhibited the contraction of the strip produced by U-50488H noncompetitively, also in a dose-dependent manner, probably without involving dopamine mechanism. The results indicated that kappa subtype is chiefly involved in the contraction of porcine coronary artery. The conclusion agrees with our autoradiography study of opioid receptor in porcine coronary artery.

[Immune function of guifudihuang pills (GFDH) and their oral liquid preparation].

Experimental observation was made on the effect of GFDH pills and their oral liquid preparation on thymus and spleen weight, formation of the antibody haemolysis plaque and phagocytic function of the macrophages of peritoneal cavity. The result revealed that both GFDH pills and their oral liquid preparation were found helpful to the enhancement of immunological function. The oral liquid preparation also increased markedly the plasma level of testosterone in immature rats.

[Phencyclidine receptors in porcine coronary artery].

By using radioligand assay, there was a phencyclidine (Phe) binding site in porcine coronary artery. This binding was specific, reversible, saturable, and stereoselective. The Scatchard analysis showed that the binding site was a single class, with dissociation constant (Kd) and maximum binding (Bmax) of 27.7 +/- 6.9 nmol.L-1 and 0.82 +/- 0.15 pmol/mg protein, respectively. The displacement experiments revealed that the [3H]Phe binding was displaced by nonradioactive Phe, TCP (Phe receptor agonists), and dextrophan (its antagonist). d-INN, a ligand of sigma receptor, had a weaker activity of displacement. These showed a dose-dependent manner. Both etorphine, an agonist of opioid receptor, and N-methyl-D-aspartic acid (NMDA), an excitatory amino acid, failed to displace the binding. These results suggest that the Phe receptors exist in the porcine coronary artery.

Effect of phencyclidine on contraction of porcine coronary vessel strips.

Classic muscular contraction experiment was used to study the effect of phencyclidine (Phe) on spiral strips of porcine coronary vessels. It showed that Phe and its analogs: 1-[1-(2-thienyl)cyclohexyl] piperidine (TCP), (+)-5-methyl-10, 11-dihydro-5H-dibenzo [a,d] cyclohepten-5, 10-imine maleatel (dizocilpine maleate) and sigma receptor ligand N-allyl-N-normetazocine (SKF10047) all exhibited a concentration-dependent contraction of porcine coronary vessel strips. Phe had no effect on the electrically stimulated contraction of the spiral strips. Dextromethorphan (Dex), a Phe receptor antagonist, and haloperidol (Hal), a sigma receptor antagonist, partially antagonized and suppressed the actions of Phe in a non-competitive manner. These results suggested that Phe, TCP, dizocilpine maleate, and (+), (-)-SKF10047 cause contraction through Phe and sigma opioid receptors.

Quaternary ammonium salt derivative of haloperidol inhibits KCl-induced calcium increase in rat aortic smooth muscle cells.

AIM: To study the effect of quaternary ammonium salt derivative of haloperidol (F3) on intracellular calcium (Ca2+) dynamic change in vascular smooth muscle cells (VSMC). METHODS: Using laser confocal scanning microscopy (LCSM), to observe the effect of F3 (0.01 - 10 micromol/L) on increase of intracellular Ca2+ fluorescent intensity induced by KCl (30 mmol/L) in VSMC. RESULTS: KCl induced a rapid increase of intracellular Ca2+ fluorescent intensity. F3 inhibited the rapid increase of that in both concentration-dependent manner (KCl: 67 +/- 24; F3 0.01 micromol/L: 57 +/- 13; 0.1 micromol/L: 40 +/- 13; 1 micromol/L: 29 +/- 9; 10 micromol/L: 20 +/- 6) and time-dependent manner; during 0 - 30 s after F3 was added, the change of fluorescent intensity was more rapid. CONCLUSION: The inhibitory effect of F3 on Ca2+ concentration in VSMC was mainly due to the blocking of the Ca2+ influx, and might partially attributed to opening of the potassium (K+) channel.

Effect of phencyclidine on dog coronary artery.

AIM: To study the effect of phencyclidine (Phe) on dog coronary artery. METHODS: Contraction of spiral strips of dog coronary artery in bioassay and coronary artery blood flow (CBF) using electromagnetic flowmeter on anesthetized dogs were observed. RESULTS: Phe 0.1-100 mumol.L-1 induced contraction of strips in a concentration-dependent manner. Dextromethorphan (Dex) 10 mumol.L-1, an antagonist of Phe receptor, antagonized the action of Phe. In vivo, Phe 10 mg.kg-1 increased flow of left circumflex coronary artery of anesthetized dogs from 334 +/- 35 mL.kg-1.min-1 to 510 +/- 58 mL.kg-1.min-1, and both left ventrical pressure (LVP) and blood pressure (BP) rose slowly after medication. Dextromethorphan (Dex) 5 mg.kg-1 also antagonized the effect of Phe. CONCLUSION: The regulation of Phe on coronary artery in vivo differs from that in vitro, which may result in the contradictory effects.