[Development and validation of a persistent postural-perceptual dizziness screening questionnaire].

Objective: To develop a simple screening questionnaire for persistent postural-perceptual dizziness (PPPD) and evaluate its screening ability. Methods: A convenience sample of 296 individuals who met the inclusion criteria between November 2021 and January 2023 were prospectively selected for three rounds of screening at the Vertigo Specialty Clinic of the Department of Otorhinolaryngology-Head and Neck Surgery in the First Hospital of Shanxi Medical University. In conjunction with expert opinion and statistical analysis, the first and second rounds of screening were used to modify and finalize the questionnaire entries, and the third round of screening was used to evaluate the questionnaire's screening ability. Independent sample t-test was used for inter group comparison, reliability and validity indicators were employed to screen and evaluate questionnaire entries, and the receiver operating characteristic (ROC) curve was plotted to determine the optimal cut-off value and corresponding sensitivity and specificity. Results: The final PPPD screening questionnaire entries included 21 items. In evaluating the reliability of this questionnaire, the Cronbach's alpha coefficient was 0.831, the half folding coefficient was 0.742, the content validity was 0.86, and the Kaiser-Meyer-Olkin (KMO) value in the structural validity was 0.811. Additionally, there were six factors with characteristic root>1 and a cumulative contribution rate of 62.62%. The area under the ROC curve of the screening questionnaire was 0.935 (95%CI: 0.877-0.992), and the optimal cut-off value was 8.5, with a sensitivity of 85.0%, a specificity of 85.5%, and a Kappa value of 0.653. Conclusion: The PPPD simple screening questionnaire designed in this study has a high sensitivity and specificity, making it a useful tool for identifying PPPD patients.

Brusatol attenuated proliferation and invasion induced by KRAS in differentiated thyroid cancer through inhibiting Nrf2.

BACKGROUND: Poorly differentiated thyroid cancer (PDTC) and anaplastic thyroid cancer (ATC) can be developed from differentiated thyroid cancer, and this dedifferentiated transformation leads to poor prognosis and high mortality. The role of Nrf2 in the dedifferentiation of differentiated thyroid cancer (DTC) induced by KRAS remains unclear. METHODS AND MATERIALS: In this study, two DTC cell lines, BCPAP and WRO, were used to evaluate the function of Nrf2 in the dedifferentiation caused by wild-type KRAS (KRAS-WT) and G12V point mutation KRAS (KRAS-G12V). RESULTS: The overexpression of KRAS-WT and KRAS-G12V increased the proliferative and invasive ability of BCPAP and WRO cells. Aggressive morphology was observed in KRAS-WT and KRAS-G12V overexpressed WRO cells. These results suggested that overexpression of KRAS-WT or KRAS-G12V may induce dedifferentiation in DTC cells. The expression of Nrf2 was increased by KRAS-WT and KRAS-G12V in DTC cells. In addition, compared with normal thyroid tissues, the expression of Nrf2 protein was considerably higher in thyroid cancer tissues on immunohistochemistry (IHC) staining, and the increased expression of Nrf2 indicated a poor prognosis of thyroid cancer. These results indicated that Nrf2 is the KRAS downstream molecule in thyroid cancer. Functional studies showed that the Nrf2 inhibitor Brusatol counteracted the proliferative and invasive abilities induced by KRAS-WT and KRAS-G12V in BCPAP and WRO cells. In addition, the xenograft assay further confirmed that Brusatol inhibits tumor growth induced by KRAS-WT and KRAS-G12V. CONCLUSION: Collectively, this study suggests that Nrf2 could be a promising therapeutic target in KRAS-mediated dedifferentiation of thyroid cancer.

Carfilzomib combined with ex vivo-expanded patient autologous natural killer cells for myeloma immunotherapy.

Natural killer (NK) cell-based immunotherapy is promising, as NK cells are in the first line of defense against cancer and capital of lysing tumor cells without pre-stimulation. However, NK cells from multiple myeloma (MM) patients are always deficient in numbers and the expression of certain activating receptors, disabling them in cytotoxicity against the cancer. Therefore, effective strategies to expand NK cells and increase NK cell-mediated cytotoxicity against MM are significant. Here, NK cells were efficiently expanded from peripheral blood mononuclear cells (PBMCs) of newly diagnosed MM patients after co-culture with irradiated K562 cells transfected with 41BBL and membrane-bound interleukin (IL)-15 (K562-mb15-41BBL) in the presence of 200 IU/ml human IL-2. The ex vivo-expanded NK cells were demonstrated to vigorously kill both MM cells and autologous primary MM cells without significant lysis of patient normal cells. Further exploration revealed a significant increase in cell surface expression of most activating receptors of NK cells and indicated that expanded NK (exp-NK) cell killing of MM cells was mediated by perforin/granzyme. NK cells are capital of lysing human leukocyte antigen (HLA) I-deficient tumor cells and carfizomib, a selective proteasome inhibitor approved for the treatment of relapsed/refractory MM patient, down-regulates the expression of HLA class I, thus enhancing NK cell-mediated lysis in MM. Here, we found for the first time that carfizomib dramatically augmented ex vivo exp-NK cell cytotoxicity against patient autologous MM cells, suggesting the use of exp-NK alone or in combination with the drug to treat MM patient.

Preoperative serum α-fetoprotein and prognosis after hepatectomy for hepatocellular carcinoma.

BACKGROUND: While the majority of studies report that a raised serum α-fetoprotein (AFP) level before operation is associated with a high risk of recurrence and death in patients who undergo hepatectomy for hepatocellular carcinoma (HCC), results are conflicting. The aim of this study was to assess the prognostic value of AFP. METHODS: Serum AFP levels were measured in patients with hepatitis-associated HCC who underwent hepatectomy between 1995 and 2012. Kaplan-Meier and multivariable analyses were performed to identify risk factors for overall and disease-free survival. Univariable and multivariable Cox proportional hazards regression was used to evaluate the predictive value of AFP. Receiver operating characteristic (ROC) curves were generated to identify the AFP level that had the highest accuracy in discriminating between survivors and non-survivors. RESULTS: Some 376 patients with hepatitis B virus (HBV)-associated HCC were included in the study. The overall survival rate was 58·8 per cent in patients with an AFP level of 400 ng/ml or less compared with 40·4 per cent for those with a level exceeding 400 ng/ml (P = 0·001). AFP concentration above 400 ng/ml was an independent risk factor for shorter disease-free and overall survival after surgery. ROC analysis indicated that the optimal cut-off values for AFP varied for different subtypes of HCC. The sensitivity and specificity were lower with areas under the ROC curve of less than 0·600. An AFP level greater than 400 ng/ml was not sensitive enough to predict the prognosis in patients with an HCC diameter smaller than 3 cm. CONCLUSION: A serum AFP level above 400 ng/ml predicts poor overall and recurrence-free survival after hepatectomy in patients with HBV-associated HCC. AFP is not a strong prognostic marker given its poor discriminatory power, with low sensitivity and specificity.

HBx mutants differentially affect the activation of hypoxia-inducible factor-1α in hepatocellular carcinoma.

BACKGROUND: Mutations in HBx gene are frequently found in HBV-associated hepatocellular carcinoma (HCC). Activation of hypoxia-inducible factor-1α (HIF-1α) contributes to HCC development and progression. Wild-type HBx has been demonstrated to activate HIF-1α, but the effect of HBx mutations on HIF-1α has not been elucidated. METHODS: HBx mutations were identified by gene sequencing in 101 HCC tissues. Representative HBx mutants were cloned and transfected into HCC cells. Expression and activation of HIF-1α were analysed by western blot and luciferase assays, respectively. The relationship between HBx mutants and HIF-1α expression in HCC tissues was also evaluated. RESULTS: The dual mutations K130M/V131I enhanced the functionality of HBx as they upregulated the expression and transcriptional activity of HIF-1α. The C-terminal truncations and deletion mutations, however, weakened the ability of HBx to upregulate HIF-1α. Meanwhile, the C-terminus was further found to be essential for the stability and transactivation of HBx. In the HCC tissues, there was a positive association between the HBx mutants and HIF-1α expression. CONCLUSION: Different mutations of HBx exert differentiated effects on the functionality of HIF-1α, however, the overall activity of HBx mutants appears to increase the expression and transcriptional activity of HIF-1α.

Hepatitis B virus x protein induces autophagy via activating death-associated protein kinase.

Hepatitis B virus x protein (HBX), a product of hepatitis B virus (HBV), is a multifunctional protein that regulates viral replication and various cellular functions. Recently, HBX has been shown to induce autophagy; however, the responsible mechanism is not fully known. In this study, we established stable HBX-expressing epithelial Chang cells as the platform to study how HBX induced autophagy. The results showed that the overexpression of HBX resulted in starvation-induced autophagy. HBX-induced autophagy was related to its ability to dephosphorylate/activate death-associated protein kinase (DAPK). The block of DAPK by its siRNA significantly counteracted HBX-mediated autophagy, confirming the positive role of DAPK in this process. HBX also induced Beclin 1, which functions at the downstream of the DAPK-mediated autophagy pathway. Although HBX could activate JNK, a kinase known to participate in autophagy in certain conditions, the change in JNK failed to influence HBX-induced autophagy. In conclusion, HBX induces autophagy via activating DAPK in a pathway related to Beclin 1, but not JNK. This new finding should help us to understand the role of autophagy in HBX-mediated pathogenesis and thus may provide targets for intervening HBX-related disorders.

Cinacalcet hydrochloride-nanoemulsion: preparation, characterization, enhanced bioavailability and pharmacodynamics.

OBJECTIVE: The aim of the study was to improve the bioavailability of Cinacalcet hydrochloride (CLC) and enhance its efficacy by the nanoemulsion drug delivery system. MATERIALS AND METHODS: First, cinacalcet hydrochloride-nanoemulsion (CLC-NE) was prepared and optimized through the pseudo ternary phase diagram and central composite design response surface methodology (CCD). The release of CLC-NE in vitro was investigated with four different dissolution media, and the bioavailability of CLC-NE in vivo was studied through beagle dogs. Finally, the pharmacodynamics of CLC-NE was evaluated by the rat model of uremia. RESULTS: Oleic acid, op-10, and PEG-200 were selected as oil phase, emulsifier, and co-emulsifier, respectively. The optimum ratio of oleic acid, op-10, PEG-200, and water was 9.87%, 38.33%, 12.78%, and 39.02%. CLC-NE has similar dissolution rates in different pH media, and the relative bioavailability of CLC-NE was 166.5%. The uremia model showed that CLC-NE could enhance renal function and reduce the excessive phosphorus (P), serum creatinine (Scr), and urea nitrogen (Urea) of model rats, as well as the inhibited increase of fibroblast growth factor-23 (FGF23) and parathyroid hormone (PTH). CONCLUSIONS: The solubility, bioavailability, and pharmacodynamics of CLC can be significantly improved through the nanoemulsion drug delivery system.

[Design and verification of the screening questionnaire for benign paroxysmal positional vertigo].

Objective: To design and validate a high-quality rapid screening questionnaire based on the common medical history and clinical experience of benign paroxysmal positional vertigo (BPPV). Methods: A questionnaire was designed based on expert's opinions, and the first-time patients who complained of dizziness and vertigo in the vertigo clinic of the First Hospital of Shanxi Medical University from September 2020 to June 2021 were prospectively screened. Taking the displacement test as the gold standard, the reliability and validity of the questionnaire were tested to evaluate its authenticity, reliability and benefit value. This study was divided into three steps. The first step was to conduct a pre-experiment and to adjust the questionnaire items; the second step was to determine the questionnaire items and the best cut-off value; the third step was to screen patients with the best cut-off value and to evaluate the quality of the questionnaire. Results: Seven items were finalized. The Cronbach's coefficient of the questionnaire was 0.675, the content validity was 0.85, the KMO value of the construct validity was 0.648, and there were 4 factors with characteristic root>1, and the cumulative contribution rate was 76.309%. The area under the receiver operating characteristic curve (ROC) was 0.938, and its optimal cut-off value was 4.5 points. At this point, the sensitivity was 88.89% and the specificity was 85.44%. Conclusion: The BPPV rapid screening questionnaire has high sensitivity and specificity, which can be used for clinical screening of BPPV patients.

Therapeutic effect of alpha-fetoprotein promoter-mediated tBid and chemotherapeutic agents on orthotopic liver tumor in mice.

Previous studies have shown that the application of Ad/AFPtBid significantly and specifically killed hepatocellular carcinoma (HCC) cells in culture and subcutaneously implanted in mice. This study was to test the therapeutic efficacy of Ad/AFPtBid in an orthotopic hepatic tumor model. Four weeks after implantation of tumor cells into the liver, nude mice were treated with Ad/AFPtBid alone or in combination with 5-fluorouracil (5-FU). Serum alpha-fetoprotein (AFP) was measured as a marker for tumor progression. The results showed that Ad/AFPtBid significantly inhibited Hep3B tumor growth. Ad/AFPtBid and 5-FU in combination was more effective than either agent alone. Tumor tissues of Ad/AFPtBid alone or combination treatment groups showed a decrease in cells positive for proliferation cell nuclear antigen, but an increase in apoptosis. Ad/AFPtBid did not suppress the hepatic tumor formed by non-AFP-producing hepatoma SK-HEP-1 cells or colorectal adenocarcinoma DLD-1 cells. The survival rate was higher in mice treated with Ad/AFPtBid plus 5-FU than those treated with either agent alone. No acute toxic effect was observed in mice receiving Ad/AFPtBid. Collectively, Ad/AFPtBid can specifically target and effectively suppress the AFP-producing orthotopic liver tumor in mice without obvious toxicity, indicating that it is a promising tool in combination with chemotherapeutic agents for treatment of AFP-producing HCC.

[Room tilt illusion: a chief complaint need to be alert in otology clinic].

Objective:This study aims to investigated the clinical significance of room tilt illusion（RTI） in screening for central vertigo in otology clinic. Method:A retrospective study of 8 RTI cases out of 656 patients with dizziness and/or vertigo. The characteristics of the symptoms, signs, physical examination, and other examinations of RTI patients were analyzed. Result:①Patients with RTI complaints accounted for 1.2% of all patients with dizziness and/or vertigo in the same period. Of all 8 patients, 2 were with acute vestibular syndrome, 5 were with episodic vestibular syndrome, and one had no dizziness and/or vertigo attack. ②Two was diagnosed with acute cerebellar infarction, and 3 was diagnosed with posterior circulation transient ischemic attacks, 2 were diagnosed with definite vestibular migraine, and one was unable to identify the cause. ③All 8 patients had a sensation of environment tilting, which was episodic（single or repeated）, transient（seconds to minutes）, in the coronal plane, and with tilt angle 30-180°. ④Seven patients had dizziness and/or vertigo for minutes to hours after a transient RTI, and one patient had no other symptoms after RTI. ⑤All patients had no decrease in vHIT gain, 6 patients had abnormal oculomotor function, and one patient had mild horizontal semicircular canal paresis. Conclusion:Patients with RTI complaints mainly had central dizziness and vertigo diseases, and once are found in otology clinic, should be alert to the possibility of suffering from central diseases.

[How to initially screen common central "malignant vertigo" at the bedside?]

Most of the common dizziness/vertigo diseases in otology clinic are peripheral vertigo diseases, and while about 20% -30% of all vertigo diseases are caused by central nervous system diseases. Pseudo vestibular neuritis, acute ischemic stroke with audiovestibular loss, and central paroxysmal positional vertigo are the three types of central "malignant vertigo" diseases which are most easily misdiagnosed by otologists. This article described the clinical characteristics and bedside physical performance of these three diseases, and summarized the differential diagnosis points for clinical reference.

[Clinical characteristics of LC-BPPV patients with bilateral direction-fixed horizontal nystagmus in primary Roll test].

Objective:To investigate the clinical features of LC-BPPV with direction-fixed horizontal nystagmus in the primary Roll test. Method:Nine patients of LC-BPPV with bilateral direction-fixed horizontal nystagmus in the first Roll test were treated by repositioning maneuvers after judging the affected side and the prognosis was observed. Result:The affected side of eight patients was the side of the apogeotropic nystagmus. Barbecue or Gufoni maneuvers was effective for reposition. The other one had no conversion of nystagmus and could not judge the affected side. Conclusion:The nystagmus performance of patients with LC-BPPV is complicated. It is necessary to carefully analyze the nystagmus performance of each position. Combining with bow and lean test, lying down test and null plane, the position of the otolith is inferred. Comprehensive use of the Barbecue or Gufoni maneuvers, forced side lying on the affected side, mastoid sniper, shaking head method is effective to complete the reposition treatment.

Haem oxygenase-1 plays a central role in NNK-mediated lung carcinogenesis.

The tobacco-specific nitrosamine, 4-(N-methyl-N-nitrosoamino)-1-(3-pyridyl)-1-butanone (NNK), is a potent lung cancer inducer. However, how NNK induces lung cancer is still largely unknown. Haem oxygenase (HO)-1 was evaluated in 30 pairs of lung cancer tumour samples and matched nontumour tissues from patients with a history of cigarette smoking. Expression of HO-1, p21(Cip1/Waf1/Cid1) (p21), B-cell lymphoma (Bcl)-2 family members, mitogen-activated protein kinase and nuclear factor (NF)-kappaB was also studied in lung cancer cells treated with NNK. The levels of HO-1 and p21 were significantly increased in lung tumour tissues. There was a positive relationship between these two proteins in the tumour. NNK stimulated lung cell proliferation and elevated the levels of HO-1, p21, inhibitor of apoptosis protein (c-IAP)2 and Bcl-2, but downregulated Bad. These effects of NNK were blocked by zinc protoporphyrin-XII, an HO-1 inhibitor. The NNK-mediated expression of HO-1 was governed by NF-kappaB and extracellular signal-regulated kinase 1/2, since blocking either of these prevented the stimulatory effect of NNK on HO-1, as well as molecules downstream of HO-1, such as p21, c-IAP2, Bcl-2 and Bad. In conclusion, haem oxygenase-1 plays a central role in NNK-mediated cell proliferation by promoting the expression of p21(Cip1/Waf1/Cid1), inhibitor of apoptosis protein 2 and B-cell lymphoma-2 but inhibiting the activity of Bad. Nuclear factor-kappaB and extracellular signal-regulated kinase 1/2 function upstream of haem oxygenase-1. Therefore, haem oxygenase-1 is likely to be a potential target in the treatment of smoking-related lung cancer.

Mechanisms of lymphocyte loss in SARS coronavirus infection.

1. Human lymphocytes and monocytes are not permissive to productive SARS coronavirus (SARS-CoV) infection in vitro. 2. Challenge of lymphocytes and monocytes with infectious SARS-CoV, inactivated virions, and receptor-binding fragment of spike protein does not trigger apoptosis. 3. Direct infection/interaction between viruses and lymphocytes/monocytes is unlikely to be the cause of lymphopaenia in SARS patients. 4. Lymphopaenia in SARS patients is likely to result from indirect mechanisms secondary to the viral infection.

[Studies on quality standard of PsL 5F injections].

OBJECTIVE: To establish the quality standard of PsL injections containing mainly 5F (ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid). METHOD: The identification of PsL was performed by thin-layer chromatography, and the content was determined by HPLC. The column was Hypersil C18 (4.6 mm x 250 mm, 5 microm), the mobile phase was the mixture of methane-water-acitic acid (55:45: 0.045) with a flow rate of 1.0 mL x min(-1), the detective wavelength was 254 nm, and the column temperature was maintained at 35 degrees C. The pH value and K+ content of the three batchs injection were determined with pH meter and flame photometric meter, and the contents of tannin, protein, oxalic acid salt and heavy metals were detected by deferent methods. RESULT: The TLC method was suitable for the identification of PsL5F. The linearity for 5F was obtained over the range of 30-240 microg x mL(-1) (r = 0.999 8), the average recovery of 5F was 99.8%. The injections were of pH value range from 7.80 to 8.20, K+ contents less than 10 mmol x L(-1), and the contents of tannin, protein, oxalic acid salt and heavy metals were qualified with the Chinese pharmacopoeia, respectively. CONCLUSION: It's sensitive and reliable that can be used as quality control methods of PsL5F injections.

[The clinical characteristics of the evoked nystagmus in the non-affected side during Dix-Hallpike test in the unilateral posterior semicircular canal benign paroxysmal positional vertigo].

Objective: To investigate the clinical characteristics of the evoked nystagmus in the non-affected side during Dix-Hallpike test(D-H test)in unilateral posterior semicircular canal benign paroxysmal positional vertigo(PC-BPPV). Method: Two hundred and thirty-six patients were diagnosed with unilateral PC-BPPV in the Tinnitus and Dizziness clinic.Among them,14 patients exhibited positive nystagmus when the non-affected side was stimulated by D-H test. The clinical data from this subgroup of patients were retrospectively analyzed. Result: The upbeat and torsional nystagmus of 14 patients were all evoked by D-H test in the affected side. 11 cases were right PC-BPPV and 3 cases were left PC-BPPV. Among these 14 patients, 5 patients exhibited the upbeat and torsional nystagmus during D-H test in the non-affected side, which was in the same direction as that evoked in the affected side. Among them, 4 cases were right PC-BPPV and 1 case was left PC-BPPV. However, the downbeat nystagmus of the remaining 9 patients were evoked by D-H test in the non-affected side, in which 7 cases were right PC-BPPV and 2 cases were left PC-BPPV.The nystagmus and vertigo of all patients completely disappeared after performing the Epley or Semont repositioning maneuvers on the affected side. Conclusion: The bilateral positive nystagmus of unilateral PC-BPPV can be evoked by D-H test. The form of nystagmus on the non-affected side is related with the initial position of otoconia in affected semicircular canal and the moving direction of otoconia during the positional test. It is effective to perform Epley or Semont repositioning maneuvers on the affected side. Meanwhile, both the effect of maneuvers and the type of nystagmus evoked by D-H test can verify which side is affected.

Single nucleotide polymorphism in the promoter region of human alpha-fetoprotein (AFP) gene and its significance in hepatocellular carcinoma (HCC).

BACKGROUND: Variations of serum AFP levels in HCC patients and cell lines are likely due to the differential activity of enhancer/silencer elements that control AFP. To understand the potential mechanism underlying the differential expression of AFP, we have examined the sequence of the AFP promoter in HCC. METHODS: Direct DNA sequencing was carried out to sequence 980 bp of AFP promoter of DNA samples isolated from 83 HCC patients. RESULTS: Three novel SNPs in the promoter region of the AFP gene, which have not been previously reported, were found at positions -330, -401 and -692. The level of serum AFP was significantly higher in HCC patients with the CT genotype of 330 SNP or the AG genotype of the 401 SNP. The genotype of CG in 692 SNP was also associated with a significant elevated level of serum AFP, and further this genotype appeared to be associated with the high risk of HCC development. 401 SNP and 692 SNP were located at the positions of known binding sites for transcription factors that have a role in the production of AFP and the growth of tumors. CONCLUSIONS: The novel polymorphisms identified in the promoter region of the AFP gene may be pathologically significant in HCC.

[Expression and significance of serum hepatocyte growth factor and myeloperoxidase in patients with obstructive sleep apnea hypopnea syndrome].

Objective:The aim of this study is to explore the expression and significance of serum HGF and MPO in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) in adults. Method:Ninety-six cases of OSAHS patients diagnosed by PSG were divided into 3 groups: Heavy, medium and light according to the range of AHI; 32 cases of outpatients were healthy persons as the control group. The serum HGF and MPO were determined by ELISA; HGF and MPO were measured after comprehensive treat with CPAP in patients with severe OSAHS, and the correlation between HGF, MPO and PSG were analyzed. Result:①With the severity of OSAHS patients increased, the serum levels of HGF and MPO increased gradually (F=119.006, 25.138; P<0.05); The concentration of HGF and MPO in each group showed that there was no significant difference between the two groups compared with the control group (P>0.05); The rest of the two groups were significantly different (P<0.05). ②There was no correlation between serum levels of HGF, MPO, BMI and age in OSAHS patients (P>0.05). The change of serum concentration was positively related to the two. All were negatively correlated with AHI, and positively correlated with LSaO2 (P<0.05). ③LSaO2 of 32 patients with severe OSAHS of has significantly increased after 3 months of comprehensive treatment, AHI and peripheral blood HGF, MPO levels were significantly reduced (P<0.05). The amylin and myeloperoxidase concentrations of blood after 6 months of treatment were significantly lower than those of 3 months after treatment (P<0.05), which still increased compared with the control group. Conclusion:Surgery combined with CPAP treatment can significantly reduce the degree of hypoxia and cardiovascular damage in OSAHS patients, which was significantly correlated with the treatment time. Combined detection of serum HGF and MPO concentrations in patients with OSAHS has a certain clinical value in judging the condition and curative effect and evaluating the cardiovascular damage.