[Preliminary study on the expression of MIF in HCC tissues and its relationship with ERK1/2 signaling pathway].

Objective: To investigate the expression of Macrophage migration-inhibitory factors (MIF) in hepatocellular carcinoma (HCC) tissues and its interaction with ERK1/2 signaling pathway, so as to establish a theoretical basis for further studying the molecular mechanism of MIF promoting HCC. Methods: From February 2020 to August 2021, 52 cases of hepatocellular carcinoma (HCC) tissues based on hepatitis B cirrhosis (HBV-LC) and 52 cases of adjacent tissues in Tianjin Medical University Cancer Hospital and 940th Hospital of Joint Logistic Support Force of PLA were collected as the experimental group, including 39 males and 13 females, aged 35-65 years. And 20 cases of normal liver tissue were selected as the control group. Immunohistochemistry was used to detect the expression of MIF, ERK1/2 and p-ERK1/2 proteins in liver tissues of the two groups, and in situ hybridization was used to detect the expression of ERK1/2 nucleic acid in liver tissues of the two groups.HepG2 HCC cells and L-02 normal hepatocytes were co-cultured with different concentrations of rMIF, the expression and phosphorylation levels of ERK1/2 and JNK1 proteins in the two kinds of liver cells were detected by Western-blot, and the expression levels of ERK1/2 nucleic acids in the two kinds of liver cells were detected by RT-PCR. One-way ANOVA was used for measurement data and χ2 test was used for counting data. Results: The expressions of MIF, ERK1/2, p-ERK1/2 and ERK1/2 mRNA were significantly increased in HCC and para-cancer tissues (the expression of MIF in HCC group was 78.8%, and that in adjacent group was 75.0%; ERK1/2 80.8% in HCC group and ERK1/2 71.8% in paracancerous group. The expression of p-ERK1/2 75.0 % in HCC group and 46.2% in paracancerous group were respectively detected. ERK1/2 mRNA was expressed in HCC group 76.9%, ERK1/2 mRNA expression in paracancerous group 78.8%), and the differences were statistically significant compared with normal liver tissues (P<0.05), but there was no significant difference between HCC and para-cancer tissues (P>0.05). The expressions of ERK1/2, p-ERK1/2 and ERK1/2 mRNA in HepG2 HCC cells were significantly increased with the increase of rMIF concentration, and the increase was most obvious when rMIF concentration was 200 ng/ml, and the difference was statistically significant compared with L-02 normal hepatocytes (P<0.05). Conclusion: MIF, ERK1/2 and p-ERK1/2 are highly expressed in HCC tissues and HepG2 HCC cells, suggesting that MIF promotes the occurrence and development of hepatocellular carcinoma through ERK1/2 signaling pathway.

[Esculin inhibits proliferation of triple negative breast cancer cells by down-regulating FBI-1].

Objective: To investigate the effect of esculin on the proliferation of triple negative breast cancer cells and its molecular mechanism. Methods: MDA-MB-231 cells were treated with 28, 56, 112, 225, 450 and 900 µmol/L of esculin for 24, 48 and 72 h, respectively, and the cell viability was detected by cell counting kit 8 (CCK-8) assay. In addition, MDA-MB-231 cells were treated with 0, 225, 450 and 900 µmol/L of esculin for 48 h. And then the changes in cell morphology were observed by inverted microscope. The clone-forming ability was detected by colony formation assay. The mRNA expression levels of FBI-1, p53 and p21 were detected using real-time fluorescence quantitative polymerase chain reaction. The protein expression levels of FBI-1, p53, p21 and Ki67 were detected by western blot. Results: Compared with the blank control group, the cell viability of MDA-MB-231 cells that treated with esculin significantly decreased in a dose-dependent and time-dependent manners. After treatment with esculin, MDA-MB-231 cells shrunk, flattened, adhered poorly to the culture dish and the cell spacing became larger. Meanwhile, shedding and incomplete cells appeared, of which 900 µmol/L of esculin treatment group showed the most dramatic changes. In addition, the colony formation ratios were decreased to (77.18±5.13)%, (65.94±4.98)% and (45.92±3.70)% in the 225, 450 and 900 µmol/L of esculin treatment groups compared with blank control, respectively (P<0.01). Furthermore, the mRNA and protein expressions of FBI-1 increased, while the levels of p53 and p21 mRNA and protein, as well as the protein expression of Ki67 decreased in a concentration-dependent manner (P<0.01). Conclusion: Esculin may regulate cell cycle-related p53-p21 pathway via FBI-1 mediated DNA replication, thus inhibit the proliferation of triple negative breast cancer cells.

Peripheral neutrophil CD64 index combined with complement, CRP, WBC count and B cells improves the ability of diagnosing bacterial infection in SLE.

OBJECTIVE: To investigate the diagnostic role of complement C3, complement C4, C-reactive protein (CRP), procalcitonin (PCT), white blood cell count (WBC), neutrophil CD64 (nCD64) index, lymphocyte subsets and their combination in differentiating bacterial infection from disease relapse in systemic lupus erythematosus (SLE). METHODS: The above biomarkers in 36 hospitalized SLE patients with bacterial infection and 45 with lupus flare without infection were retrospectively studied. Bacterial infection was proven by positive cultures or typical clinical symptoms and signs combined with positive response to antibiotics. Lupus flare was defined as three points greater than their previous SLE disease activity index score. The diagnostic value for bacterial infection was evaluated by the areas under the receiver operating characteristic curves (AUC) and a novel bioscore system combining multiple biomarkers. RESULTS: Increased CRP ( p = 0.049), WBC ( p = 0.028) and nCD64 index ( p = 0.034) were observed in the infected group and C3 ( p = 0.001), C4 ( p = 0.016) and B cells levels ( p = 0.010) were significantly reduced. The AUC for the above six biomarkers had no significant difference. Interestingly, the combination of nCD64 index, CRP, WBC, C3 and C4 improved significantly the diagnostic potential of SLE infection (AUC 0.783 (interquartile range 0.672, 0.871), p < 0.001; sensitivity 85.29% specificity 62.50%). In the bioscore system including the above six biomarkers, the bacterial infection rate in patients with bioscore ≤2, 3, 4, 5 and 6 were 0.00, 39.29, 59.10, 61.54 and 100.00%, respectively. CONCLUSION: The combination of nCD64 index, C3, C4, CRP, WBC and B cells in a bioscore is useful to diagnose bacterial infection in SLE.

[A clinical study on the pathogenesis of lung cancer-related cerebral infarction].

Objective: To explore the risk factors for lung cancer-related cerebral infarction. Methods: The hospitalized active lung cancer patients on anti-cancer therapy with no traditional stroke risk factors, who experienced an acute cerebral infarct in the First Affiliated Hospital of Guangxi Medical University from January 2005 to December 2015, were consecutively collected as the LCRS (lung cancer-related stroke) group. The active lung cancer patients without cerebral infarction hospitalized at the same peroid matched with the LCRS group for age and gender were collected as the LC (lung cancer) group. Clinical data from the two groups were analyzed. Results: A total of 139 LCRS patients and 139 LC patients were enrolled in the study, with 110 male and 29 female in each group, and there were no significant difference for the mean age between the LCRS group (52.1±10.4 years old ) and the LC group (52.1±10.1 years old). Two or more acute ischemic lesions of the brain were showed by MRI in most patients in the LCRS group (117 cases, 84.2%). Compared with the LC group, more patients in the LCRS group were found with adenocarcinoma, metastasis, elevated plasma D-dimer, CA125 and CA199 levels [88 cases (63.3%) vs 47 cases (33.8%); 98 cases (70.5%) vs 56 cases (40.3%); (468.38±291.37) µg/L vs (277.59±191.22) µg/L; (221.42±146.34) U/ml vs (106.84±69.97) U/ml; (254.68±185.84) U/ml vs (97.15±63.64) U/ml; with all P<0.001]. By logistic regression analysis of multiple factors, the elevated plasma D-dimer, CA125 and CA199 levels were showed to be independent risk factors for the cerebral infarction (OR=1.003, 95%CI 1.001-1.004; OR=1.006, 95%CI 1.003-1.010; OR=1.011, 95%CI 1.007-1.015). Conclusions: The elevated plasma D-dimer, CA125 and CA199 levels are the risk factors for the lung cancer related cerebral infarction, which may lead to hypercoagulation and induce cerebral infarction eventually.

[Clinical analysis of idiopathic sudden sensorineural hearing loss with vertigo and without vertigo].

Objective:To explore the clinical characteristics and treatments of patients with idiopathic sudden sensorineural hearing loss（ISSHL） with or without vertigo. Method:One hundred and twenty ISSHL cases were divided into vertigo group (n=36) , without vertigo group (n=84) , and with benign paroxysmal positional vertigo group (n=15). All patients were in regular treatment. Besides, according to the types of BPPV, patients do the Epley maneuver or Barbecue roll maneuver. We summarized the result and treatment of the patients. Result:The audiometric curve of ISSHL with vertigo were mainly at flat type. After treatment of the ISSHL patients were better than the patients with vertigo in the degrees of hearing loss . Furthermore, the rate of the patients of marked efficiency, efficiency and total efficiency of ISSHL was lower than the ones without.The patients with BPPV, including 12 cases of posterior semicircular canal and the 3 cases of lateral semicircular canal, were all ipsilateral. Conclusion:ISSHL with vertigo group lost hearing is severer than ISSHL without vertigo. Thus the hearing and the treatment effect were worse.The symptoms without vertigo in ISSHL were better than the patients with vertigo.

[Pediatric laryngeal clefts: an experience in the diagnosis and management of 13 cases].

Objective: To investigate the diagnosis and management of laryngeal cleft. Method: The clinical data of 13 cases of laryngeal cleft treated between 2007 and 2015 was analyzed retrospectively. Results: The children with laryngeal cleft were classified according to the classification of Benjamin-Inglis, as type Ⅰ(11 cases), typeⅡ(1 case) and type Ⅲ(1 case). All patients were confirmed by microlaryngobronchoscopy under general anaesthetic. Eleven typeⅠ and 1 type Ⅱ clefts were managed conservatively, with which all type Ⅰ patients were successfully managed, while the type Ⅱ patient was resolved by surgical endoscopy. The type Ⅲ patient was treated by open repair but the results was poor. Conclusions: Patients who suffered with choking on feeding or recurrent aspiration pneumonia, especially coexisted with other congenital malformation, needed detailed evaluation for laryngeal cleft, although which was a rare congenital abnormality. Electronic laryngoscope could be the first step to screen the cleft, while microlaryngobronchoscopy is the gold standard for diagnosis of laryngeal cleft. The majority of children with lower type clefts can be managed conservatively. Surgical endoscopy has high success rate when strictly following the indication. Type Ⅲ and Ⅳ clefts have high mortality for usually combining with severe complications and abnormalities.

[Clinical analysis of benign paroxysmal positional vertigo associated with Meniere's disease].

Objective:To explore treatment and therapeutic effectiveness of patients with Meniere' s disease and benign paroxysmal positional vertigo.Method: A series of BPPV 60 cases was retrospective analyzed. The patients were divided into three groups: Meniere' s disease with multiple semicircular canal BPPV(n=6), with single semicircular canal BPPV (n=11) and BPPV without Meniere' s disease group(Control,n=43). All patients were diagnosed by the Dix-Hallpike test or roll test and treated with the canalith repositioning procedure. The outcomes were compared among the three groups. Result: Unilateral semicircular canal BPPV was more than bilateral BPPV, the posterior semicircular canal was the most common canal involved, and multiple semicircular canal BPPV with Meniere' s disease patients needed repeated canalith repositioning procedure and had a higher recurrence rate.Conclusion:A lower success rate of treatment and a higher recurrence rate was found in BPPV patients with Meniere' s disease compared with those without Meniere' s disease . The recurrence rate is highest in multiple semicircular canal BPPV with Meniere' s disease.

Physiological basis for long life span.

A collection of clinical data is reported on nonagenarians in comparison to an 'average' population of younger age. The results of these clinical data indicated that a vital physiological basis for long life span probably existed. The basis include a better micro-blood-flow state, a better cardiac, immune (nature killer cell activity), adrenocortical, hepatic and renal function, and a higher level of high density lipoprotein cholesterol. It is suggested that the method, including Chinese traditional medicine, to improve the micro-blood-flow, nature killer cell activity, high density lipoprotein cholesterol and vital organ function may be beneficial for life preservation and aging retarding.

A comparative study of 632.8 and 532 nm laser irradiation on some rheological factors in human blood in vitro.

The effects of laser irradiation with 632.8 and 532 nm on rheological properties of blood were comparatively studied in vitro. Under the irradiation condition of 30 mW, laser irradiation of blood samples using a spot diameter of 5 mm with each laser, showed promising results in the modulation of hemorheological properties. When blood samples from patients with abnormally high values of erythrocyte sedimentation rate (ESR) were irradiated, the values of ESR were lowered statistically by either of the 632.8 or 532 nm lasers. The laser irradiation reduced blood viscosities at different shear rates (10-110 S(-1)) for the hyper-viscosity blood samples. Laser irradiation increased the electrophoretic mobility (EPM) of erythrocytes when the values of the sample's EPM were abnormally slow. The erythrocyte deformability was enhanced by laser irradiation when the deformability of the sample from the patients was originally poor. For verifying the improvement of laser irradiation on erythrocyte deformability, the typical erythrocyte samples with poor deformability were produced by the pre-treatment of the erythrocytes with Ca(2+). The deformability of these erythrocyte samples was also improved after laser irradiation. These results suggest that membrane-bound hemoglobin (Hbm) might be the initial site of the interaction, since Hbm is the main cause of poor deformability when erythrocytes were treated with Ca(2+). In all experiments including ESR, blood viscosity, EPM and erythrocyte deformability, the 532 nm laser demonstrated more efficient effects on modulating rheological properties than 632.8 nm laser. This wavelength effect is consistent with the absorption spectrum of hemoglobin, reflecting that hemoglobin may be one of the action targets under laser irradiation.

Extended parietal cell vagotomy in the treatment of perforation, hemorrhage and stenosis due to duodenal ulcer.

Ninety-five patients with perforation, hemorrhage or stenosis due to duodenal ulcer were treated by extended parietal cell vagotomy. Postoperative follow-up ranged from 3.5 to 10 years (mean 6 years) in 88 patients (92%) with acute perforation (60), hemorrhage (8) and stenosis (20). There was no operative mortality. Ulcer recurrence was 2.3%. Only one patient (5%) had restenosis and required reoperation. There was no recurrent hemorrhage and there were few long-term complications. According to the Visick classification, 67 patients (76%) belonged to grade I, 13 (14.7%) grade II, 4 (4.5%) grade III, and 4 (4.5%) grade IV. Extended parietal cell vagotomy proved to be safe with excellent results, low ulcer recurrence and few complications. Moreover, recurrent ulcers healed rapidly following medical therapy. The authors believe that extended parietal cell vagotomy should be the treatment of choice for acute perforation, hemorrhage or stenosis due to duodenal ulcer.

[Microinjection of 5-HT into the rostral ventrolateral medulla reduced the hyperviscosity and elevation of blood pressure induced by stress].

Experiments were carried out on 62 wistar rats. The hyperviscosity and elevation of blood pressure were induced by hanging and restraining the rats with their four limbs tied on a frame. It was found that microinjection of 5-HT (25 micrograms/10 microliters) into the 4th ventricle of the brain or bilateral microinjection of 5-HT (4 micrograms/0.5 microliters/site) into rostral ventrolateral medulla (rVLM) reduced stress-induced hyperviscosity (p < 0.01) and elevation of blood pressure (p < 0.01). The effect of 5-HT injected into the 4th ventricle or rVLM was blocked by bilateral microinjection of cinanserine (4 micrograms/0.5 microliter/site) into rVLM. These results suggest that microinjection of 5-HT into 4th ventricle and rVLM could reduce stress-induced hyperviscosity and elevation of blood pressure and these effects were probably mediated via 5-HT receptors in the rVLM.

[Inhibitory effect of intraventricular administration of morphine on the hyperviscosity and elevation of blood pressure induced by stress in the rat].

Experiments were performed on 99 Wistar rats. It was found that hyperviscosity and elevation of blood pressure (BP) could be induced by hanging and restraining conscious rats with their four limbs tied on a frame. These effects were unaffected by bilateral vagotomy. By intravenous injection of propranolol or phentolamine, elevation of BP could be reduced, while stress-induced hyperviscosity could only be reduced by propranolol (i.v.). Stress-induced hyperviscosity and elevation of BP could be inhibited by electroacupuncture applied to the right hind leg or microinjection of morphine into 4th ventricle of the brain. On the other hand, if opiate receptor antagonist naloxone was given into the 4th-ventricle, the stress-induced hyperviscosity and elevation of BP could no longer be inhibited by electroacupuncture. It is suggested that the hyperviscosity and elevation of BP induced by hanging and restraining are mediated by excitatory cardiovascular sympathetic outflow with the result of activation of adrenoreceptors. Activation of the opiate receptors in the hindbrain may be responsible for decrease in stress-induced hyperviscosity and elevation of BP and for the inhibitory effect of electroacupuncture of the right hind leg on stress-induced hyperviscosity and elevation of BP.

[Effect of acute hypoxia on blood viscosity, red blood cell deformability and the left ventricular function in rats].

Experiments were performed on SD rats anaesthetized with urethane and chloralose. The left ventricular function and the parameters of hemorrheology during acute hypoxia were investigated. It was found that during acute hypoxia of 15 min, arterial blood gas value PaO2 decreased markedly. The parameters of the left ventricular function such as +/- dP/dtmax, Peak, HR, Vce40, Vmp, Vmax, L0 (CFU), decreased markedly either. At the same time, hyperviscosity was induced and red blood cell filtration (IF) increased. All these parameters recovered to normal control level after reoxygenation of 15 min. By intravenous injection of propranolol (0.5 mg/kg) and then hypoxia, the left ventricular function were suppressed markedly; and so were to the elevation of blood viscosity and IF. Preinjection of regitine (3 mg/kg) prior hypoxia has the same effect on the blood viscosity and IF. Acute hypoxia induced hyperviscosity and elevation of IF could be suppressed by destruction of carotid sinus region with phenol, but the left ventricular function decreased markedly. These results suggest that acute hypoxia induces hyperviscosity and decreases of red blood cell deformability and left ventricular function, which is probably mediated via the sympathetic nervous system and carotid sinus region.

[Effects of tetramethylpyrazine and ferulic acid alone or combined on vascular smooth muscle, blood viscosity and toxicity].

The experiments showed that both tetramethylpyrazine and ferulic acid relaxed the norepinephrine-induced spasmodic contraction of rabbit and rat aorta strips, increased the coronary flow of isolated guinea pig hearts and reduced the whole blood viscosity in rats. Evaluated with Burgi's equation, the combined effect of these 2 drugs was obviously potentiated, but the combined acute toxicity in mice was greatly reduced.