Objective　To improve the understanding and provide a reasonable plan for clinical diagnosis and treatment of TRE(TSC-related epilepsy,TRE),by analyzing the clinical and genetic characteristics of TRE as well as the treatment effect about medication and surgery.Methods　We analyzed the video EEG,imaging,pathology,genetic characteristics,and treatment effect of 10 patients retrospectively.These patients with refractory epilepsy were diagnosed and treated by our epilepsy center from November 2014 to November 2019.The clinical and genetic characteristics of TRE were summarized.Results　There were 3 females,7 males,4 patients with seizures before 3 years old,5 patients with seizures from 3 to 6 years old and 1 at 19 years old.There were 5 cases of spastic seizures and 5 cases of focal seizures.The Video EEG showed that 7 patients have epileptiform discharge site located on one side brain hemisphere and 3 patients about both side.Imaging results showed 4 cases of unilateral subventricular or subcortical nodules,5 cases of bilateral abnormalities,and 1 case of Dandy-Walker deformity.Pathology showed brain tissue section of 4 patients with six-story structure disappearance and immature neurons appearance.Genetic testing revealed 5 cases of TSC1 mutations,5 cases of TSC2 mutations,7 cases of family mutations,and 4 cases of surgical patients were nonsense mutations.The treatment effect about 3 patients of Rapamycin improved than before significantly.Conclusion　TRE patients usually fall ill in early age,more males than females,mainly spastic and focal seizures.The Video EEG showed epileptiform discharge site located on one side brain hemisphere.Gene testing is useful for treatment.The type of mutation may be related to the degree and location of brain lesions and the origin of the discharge,but there is no significant correlation between the discharge site recorded on the EEG and the abnormal imaging site.Therefore,genetic testing should be considered when clinical treatment is not improvement.

BACKGROUND: Hydroxyapatite-geltin composite has good biocompatibility and osteoinductivity, and can used be as tissue engineering scaffold to repair bone defects. OBJECTIVE: To observe the effect of three-dimensional (3D) printed hydroxyapatite/gelatin scaffold combined with bone marrow mesenchymal stem cells and umbilical vein endothelial cells in repairing rabbit skull defects. METHODS: Nine male New Zealand white rabbits were taken to establish a skull defect model with a diameter of approximately 0.8 cm, and randomly divided into three groups: blank group: no any treatment; control group: only 3D printed hydroxyapatite-geltin scaffold; experimental group: 3D printed hydroxyapatitegeltin scaffold and bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells complex group, with three rabbits in each group. At 8 weeks after the operation, CT scan of the skull pyramid beam and histological observation of the skull defect were performed on the white rabbits of each group. Animal experiments were approved by the Ethics Committee of Jining Medical College. RESULTS AND CONCLUSION: (1) CT scan of pyramidal tract: Blank group showed obvious bone defects, and the defect area was slightly radiopaque with the edges of the surrounding normal bone tissue. Control group showed some new bone formation, which was discontinuous and inconsistent with the surrounding bone tissue. Experimental group showed that the new bone tissue was linear and continuous; the thickness was thin; and the defect area merged with the adjacent bone tissue edge. (2) Histological observation: Hematoxylin-eosin staining and Masson trichrome staining showed that fibrous connective tissue formation and a small amount of free bone cells were seen in the defect area of the blank group. A small amount of bone formation was seen in the control group. Bone matrix was deposited at the edge of the material, replacing the material to form small bone trabeculae. The material space of the experimental group was gradually replaced by new bone, and the defect area was filled with new bone and trabecular bone structure-like tissue. (3) The results show that the 3D bionic printing hydroxyapatite-geltin scaffold combined with bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells can effectively promote the growth of bone tissue and accelerate the repair of bone defects.

AIM: To study the effects of small-incision lenticule extraction(SMILE)and femtosecond laser-assisted excimer laser in situ keratomileusis(FS-LASIK)on early postoperative corneal higher order aberrations(HOAs)and visual quality in patients with high myopia.

METHODS:Totally 102 patients(204 eyes)with high myopia who underwent corneal refractive surgery between August 2018 and August 2020 in the hospital were selected as study subjects for the prospective study, and they were randomly divided into SMILE group(51 cases, 102 eyes)and FS-LASIK group(51 cases, 102 eyes). The postoperative visual acuity, corneal HOAs and objective visual quality were compared between the two groups, and the correlation between corneal HOAs and objective visual quality was analyzed.

RESULTS: At 1mo after surgery, the spherical equivalent degrees in SMILE group and FS-LASIK group were significantly reduced(P<0.05), and there was no statistically significant difference in the proportion of patients with postoperative uncorrected visual acuity≥preoperative best corrected visual acuity between the two groups(95.1% vs 92.2%, P>0.05). The corneal HOAs in the two groups were significantly increased(P<0.05), and the overall spherical aberration(SA), trefoil and higher HOAs in SMILE group were lower than those in FS-LASIK group(all P<0.05). The objective scattering index(OSI)was significantly increased in the two groups while the MTF cutoff frequency(MTF_cutoff), Strehl Ratio(SR)and contrast visual acuity(VA100%, VA20%, VA9%)were significantly decreased(all P<0.05). The OSI of SMILE group was lower than that of FS-LASIK group while MTF_cutoff and VA9% were higher than those of FS-LASIK group(all P<0.05). The corneal HOAs in patients with high myopia were negatively correlated with MTF_cutoff and were positively correlated with OSI(P<0.05).

CONCLUSION:Both SMILE and FS-LASIK can effectively correct high myopia, and SMILE has a smaller increase in early postoperative corneal HOAs, and it is of great significance to relieve visual quality impairment.

The present study was aimed to investigate the effect of Janus kinase 3 (JAK3) on the migration of breast cancer cells and the underlying mechanism. The expression of JAK3 in breast cancer MCF-7 cells was silenced by siRNA (siJAK3). The migration ability of MCF-7 cells was detected by scratch test. The activity of store-operated calcium channel (SOCC) was detected by fluorescence calcium imaging. The expression levels of Orai1 and STIM1, key molecules in the process of store-operated calcium entry (SOCE) were detected by Western blot and RT-PCR. The results showed that 2-APB, an inhibitor of SOCC, could inhibit the migration ability of MCF-7 cells. siJAK3 transfection significantly inhibited the migration ability of MCF-7 cells, decreased the activity of SOCC, and down-regulated mRNA and protein expression levels of Orai1 and Stim1. Over-expression of Orai1 or STIM1 in JAK3-silenced cells restored their migration ability. These results suggest that JAK3 facilitates the migration of breast cancer cells by SOCC.
Breast Neoplasms ; enzymology ; Calcium ; metabolism ; Calcium Channels ; metabolism ; Cell Movement ; physiology ; Gene Expression Regulation, Neoplastic ; Humans ; Janus Kinase 3 ; genetics ; metabolism ; MCF-7 Cells ; ORAI1 Protein ; genetics

Objective To investigate the characteristics of voice changes by multi-dimensional acoustic analysis in patients who underwent thyroid gland lobectomy without laryngeal nerve injury.Methods To observe vocal cord movement and evaluate voice quality,32 patients with unilateral thyroid adenoma received the examinations of fiberlaryngscopy and Multi-Dimensional Voice Program before and after operation,respectively.Forty subjects with normal voice were used as cantrol group.Results Fiberlaryngscopy examinations were normal in all patients before and after surgery.In thyroid group,the preoperative acoustic parameters in male patients showed no significant difference(P＞0.05) compared with the control group.There was no significant difference between preoperative and postoperative acoustic parameters(P＞0.05).While the postoperative acoustic parameters of female patients was significantly lower than those of preoperative and in control group(P＜0.05).There was significant improvement 3 months after the operation compared with that of preoperative group(P＜0.05).Fhi of 3 months after operation was still lower than those of preoperation and in control group in male and female patients that continues to be significant after 3 months.Conclusion Voice changes may occur after unilateral thyroid gland lobectomy without any laryngeal nerve injury,and there is little voice change in male patients,while in female patients,voice improvement is observed after operation.Fhi still remaines lower level for a certain of time.

OBJECTIVE: To observe the expressions of sphingosine kinase 1 (SphK1) and sphingosine-1-phosphate receptor 2 (S1PR2) in hippocampus of epileptic rats and to investigate the pathogenesis of SphK1 and S1PR2 in epilepsy. METHODS: One hundred and eight male Sprague-Dawley (SD) rats were randomly divided into control group (n=48) and pilocarpine (PILO) group (n=60). A robust convulsive status epilepticus (SE) was induced in PILO group rats by the application of pilocarpine. Control group rats were injected with respective of physiological saline. Pilocarpine group was randomly divided into 6 subgroups (n=8): acute group (E6 h, E1 d, E3 d), latent group (E7 d) and chronic group (E30 d, E56 d). Each subgroup has 8 control rats and 8 epileptic rats. Hippocampal tissue and brain slices were obtained from control rats and rats subjected to the Li-PILO model of epilepsy at 6 h, 1 d, 3 d,7 d,30 d and 56 d after status epilepticus (SE). Western blot technique was used to determine the expressions of SphK1 and S1PR2 in hippocampus at different point of time after pilocarpine treatment. Immunofluorescence was applied to detect the activation and proliferation of hippocampal astrocytes and the localization of SphK1 and S1PR2 in rat hippocampal astrocytes. RESULTS: Compared with control group, the levels of SphK1 in acute phase (E3 d), latent phase (E7 d) and chronic phase (E30 d, E56 d) were significantly increased while the expressions of S1PR2 were decreased in acute phase (E3 d), latent phase (E7 d) and chronic phase (E30 d, E56 d)(P＜0.05 or P＜0.01). Immunofluorescence results showed astrocyte activation and proliferation in hippocampus of epileptic (E7 d) rats (P＜0.05). Confocal microscopy confirmed the preferential expressions of SphK1 and S1PR2 in epileptic rat(E7 d)hippocampal astrocytes. CONCLUSION The results indicate that SphK1 and S1PR2 may play an important role in the pathogenesis of epilepsy by regulating the activation and proliferation of hippocampal astrocytes and altering neuronal excitability.
Animals ; Astrocytes ; enzymology ; Epilepsy ; enzymology ; physiopathology ; Hippocampus ; cytology ; enzymology ; Male ; Phosphotransferases (Alcohol Group Acceptor) ; metabolism ; Pilocarpine ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Lysosphingolipid ; metabolism

To analyze the characteristics of the results of serum immunological tests and brain CT image examinations of atypical cerebral cysticercosis patients, so as to provide the reference for improving the diagnosis of the disease.Totally 446 suspected cerebral cysticercosis patients were chosen as the study objects, all of them were given experimental treatment with praziquantel, and then the patients with atypical cerebral cysticercosis were diagnosed according to the treatment effect and review results of brain CT or MRI. Meanwhile, all the 446 patients were tested for serum specific IgG and IgG4 antibodies and cysticercus circulating antigen (CAg) by ELISA, McAb ELISA and PEG-ELISA respectively, and the IHA test was also performed. All the patients received the brain CT examinations. The test results were analyzed statistically and the test performances of the methods above-mentioned were calculated.Among the 446 suspected cerebral cysticercosis patients, after the praziquantel treatment, there were 315 patients whose symptoms were alleviated, and they were diagnosed as atypical cerebral cysticercosis. Among the 446 suspected cerebral cysticercosis patients, the positive rates of specific IgG and IgG4 antibodies, and CAg were 15.47%, 15.02%, and 11.21% respectively, and the positive rate of IHA was 28.47% (χ² = 52.45, P ＜ 0.01). The brain CT examinations showed that there were 79.14% (353/446) of patients with suspected cysticercus foci. The sensitivities of ELISA, McAb-ELISA, PEG-ELISA, IHA test and brain CT examination (suspected cysticercus foci) for the diagnosis of atypical cerebral cysticercosis were 15.36%–96.82%, the specificities were 63.36%–99.24%, the positive predictive values were 86.40%–98.52%, the negative predictive values were 32.83%–98.25%, the positive likelihood ratios were 2.64–27.86, the negative likelihood ratios were 0.05–0.81, and the OR values were 7.16–52.80. The consolidation of the five tests above-mentioned showed the OR value was 108.00, which was 2 times of the OR value of CT examination.Atypical cerebral cysticercosis is definitely diagnosed mainly by means of evaluating the effect of the diagnostic therapy (anti-cysticercus), and the serum immunological examinations and brain CT examination also have important reference values.

Objective: To investigate the expression characteristics of Tim-3 on natural killer (NK) cells of peripheral blood in patients with acute myeloid leukemia (AML) and its clinical significance. Methods: Peripheral blood was obtained from 39 patients with newly diagnosed AML before intervention, with peripheral blood from 28 cases of healthy volunteers collected as normal control. Using CD3, CD56 and Tim-3 as markers, expression levels of Tim-3 on the peripheral blood NK cells were detected by immune fluorescence labeling and flow cytometry. Results: The ratio of the peripheral blood CD3(-)CD56(+) NK cells in newly diagnosed AML patients (5.74±5.31) %decreased significantly, compared with the normal control (12.55±6.33) % (t=4.596, P<0.001) . Tim-3 expression on the peripheral blood NK cells in newly diagnosed AML patients (42.67±19.08) % decreased significantly, compared with the normal control group (60.99±20.69) % (t=3.781, P<0.001) . CD3(-)CD56(+)NK cell ratio of peripheral blood in AML patients was significantly correlated with Chromosome karyotype (t=2.915, P<0.005) . Expression level of Tim-3 on NK cells in the peripheral blood of AML patients had significant correlation with ratio of CR and NCCN high risk group (P<0.05) . Conclusion: The rate of NK cells in peripheral blood and the expression level of Tim-3 on NK cells in AML patients decreased significantly.The lower expression level of Tim-3 on NK cells correlate with prognosis of AML.
Flow Cytometry ; Hepatitis A Virus Cellular Receptor 2/metabolism* ; Humans ; Killer Cells, Natural ; Leukemia, Myeloid, Acute ; Prognosis

Objective To compare broth microdilution and agar dilution methods for in vitro testing of activities of fluconazole,ketoconazole and itraconazole against clinical Malassezia isolates.Methods Broth microdilution and agar dilution methods were used to determine the minimal inhibitory concentration(MIC)of fluconazole,ketoconazole and itraconazole for 27 clinical strains(5 species)of Malassezia.Results The minimal inhibitory concentration(MIC)ranges of fluconazole,ketoconazole and itraconazole were 0.25-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.125 mg/L respectively as shown by broth microdilution method,2-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.25 mg/L respectively as revealed by agar dilution method.Both methods demonstrated that itraconazole possessed the strongest activity against Malassezia species,followed by ketoconazole and fluconazole.The agreement rate in MICs between the two methods was 78.8％,85.2％ and 88.9％,respectively for fluconazole,ketoconazole and itraconazole,with the intraclass correlation coefficients (ICCs)being 0.88,0.80 and 0.76 respectively.Conclusions Fluconazole,ketoconazole and itraconazole are highly active against Malassezia species in vitro,and itraconazole is the most active.Broth microdilution and agar dilution method coincide well in,and are applicable for,the antifungal susceptibility testing of Malassezia species in vitro.

OBJECTIVETo search for an effective method for treatment of obstinate hiccup.

METHODSNinety cases of obstinate hiccup secondary to cerebral diseases were randomly divided into 3 groups, 30 cases in each group. Group A were treated by injection of compound chlorpromazine into Yintang (Ex-HN 3), group B by intramuscular injection of compound chlorpromazine and group C by acupuncture at Yintang (Ex-HN 3). Their therapeutic effects were compared.

RESULTSThe effective rate was 93.3% in the group A, 10.0% in the group B and 30.0% in the group C, the group A being better than the group B and the group C (P < 0.01).

CONCLUSIONInjection of compound chlorpromazine into Yintang (Ex-HN 3) has a significant therapeutic effect on obstinate hiccup.