Preliminary observation of thyroid function changes in subclinical thyroid diseases.

BACKGROUND: It is estimated that 1.95% and 5.55% of adults in China suffer from subclinical thyroid diseases, which is difficult to diagnose and treat. OBJECTIVE: This study aimed to explore the development and prognosis of subclinical thyroid diseases to provide a reference from our single center experience. METHODS: A total of 240 cases from April 2019 to August 2021 in the laboratory information system database of Huanghua Development Boai Hospital were retrospectively analyzed. Binary logistic regression was conducted to analyze odds ratio (OR) of subclinical thyroid disease types returning to a normal state. RESULTS: Among the patients hypothyroidism Ia and hyperthyroidism Ia were the most common type with conversion to the normal state (P< 0.001). TSH level of patients with conversion to a normal state was significantly lower than that of those who developed to abnormal disease (P= 0.015). The OR values of hyperthyroidism Ia and hypothyroidism Ia that returned to a normal state compared with hyperthyroidism Ib were 2.659 (1.159 ∼ 6.096, P= 0.021) and 3.138 (1.1.278 ∼ 7.709, P= 0.013), respectively. The OR value of hypothyroidism Ib that returned to normal compared with hyperthyroidism Ib was 0.629 (0.131 ∼ 3.010, P= 0.561). Thyroid hormone levels, age, and gender at first diagnosis were not impact factor for prognosis of subclincal thyroid disease (P> 0.05). CONCLUSION: Cases with grade hypothyroidism Ia and hyperthyroidism Ia are more likely to revert to normal state than other subclinical thyroid diseases. TSH reference range should be explored for diagnosis and treatment.

Effects of Dezocine-Remifentanil Intravenous Anaesthesia on Perioperative Signs, Serum TNF-&aipha; and IL-6 in Liver Cancer Patients undergoing Radiofrequency Ablation.

OBJECTIVE: To analyse the impact of dezocine-remifentanil intravenous anaesthesia on perioperative signs, serum tumour necrosis factor-α (TNF-α), and interleukin-6 (IL-6) in liver cancer patients undergoing radiofrequency ablation (RFA). STUDY DESIGN: An experimental study. PLACE AND DURATION OF STUDY: Renmin Hospital of Wuhan University, Wuhan, China, from January 2017 to February 2018. METHODOLOGY: Eighty patients with small hepatocellular carcinoma (SHCC) were selected as the research object. They were divided into Group A and Group B with the random number table method, with 40 cases in each group. Group A were given dezocine-remifentanil intravenous anaesthesia and Group B were given midazolam-remifentanil intravenous anaesthesia. Patients' situations in the surgery were compared between the two groups. Changes in heart rate (HR), mean arterial pressure (MAP) and blood oxygen saturation (SpO2) were recorded before the surgery (T0), at 5 minutes after the RFA (T1) and at the end of the RFA (T2). Levels of tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) on the 12 day after the RFA were compared between the two groups. RESULTS: The wake-up time in Group A was shorter than Group B (p<0.001), and the VAS pain score in Group A was lower than Group B (p<0.001). At T1, the MAP in Group A was higher than Group B (p<0.001). There was no significant difference in MAP between the two groups at T0 and T2 (p=0.881, 0.696, respectively). At T1 and T2, the HR in Group A was lower than Group B (all p<0.001). There was no significant difference in HR between the two groups at T0 (p=0.684). There was no significant difference in SpO2 between the two groups at T0, T1 and T2 (p=0.654, 0.884 and 0.798, respectively). On the 1st day after the RFA, the level of TNF-α, IL-6 in Group A were lower than those of Group B (all p<0.001). There was no significant difference in the incidence of intraoperative complications between the two groups (p=0.644). CONCLUSION: Compared with midazolam-remifentanil intravenous anaesthesia, the dezocine-remifentanil method has a better analgesic effect, shorter wake-up time, and can effectively regulate the expression of inflammatory cytokines TNF-α and IL-6. However, the effect of remifentanil on the respiratory function is dose-dependent. Therefore, respiratory cycle monitoring and management should be strengthened during the surgery.

Overexpression of miR-381 relieves neuropathic pain development via targeting HMGB1 and CXCR4.

MicroRNA are significant regulators of neuropathic pain development. Neuroinflammation contributes a lot to the progression of neuropathic pain. miR-381 is involved in various pathological processes. However, the role of miR-381 in neuropathic pain development remains barely understood. Therefore, in our study, we aimed to investigate the effects of miR-381 on the process of neuropathic pain progression by establishing a rat model using chronic sciatic nerve injury (CCI). Here, we observed that miR-381 was dramatically decreased in CCI rats. Up-regulation of miR-381 strongly reduced neuropathic pain behaviors including mechanical and thermal hyperalgesia. In addition, inflammatory cytokine expression, including IL-6, IL-10 and TNF-α were significantly repressed by overexpression of miR-381. High mobility group box 1 protein (HMGB1) and Chemokine CXC receptor 4 (CXCR4) participate in neuropathic pain development. In our present study, HMGB1 and CXCR4 were predicted as direct targets of miR-381 by employing bioinformatics analysis. Overexpression of miR-381 was able to restrain the expression of HMGB1 and CXCR4 greatly. The direct correlation between HMGB1 and CXCR4 and miR-381 was confirmed in our research. Furthermore, we found that HMGB1 and CXCR4 were increased in CCI rats time-dependently. Moreover, it was demonstrated that silence of HMGB1 and CXCR4 in CCI rats depressed neuropathic pain progression greatly. In conclusion, it was indicated that miR-381could inhibit neuropathic pain development through targeting HMGB1 and CXCR4.

Effect of ozone oxidative preconditioning on oxidative stress injury in a rat model of kidney transplantation.

The aim of the present study was to investigate the protective effect of ozone oxidative preconditioning (OOP) on renal oxidative stress injury in a rat model of kidney transplantation. Thirty-six male Sprague Dawley (SD) rats were randomly divided into three groups: A sham (S) group, a kidney transplantation (KT) group and an OOP and kidney transplantation (OOP+KT) group. In the S group, the rats' abdomens were opened and closed without transplantation. In the KT group, the rats received a left kidney from donor SD rats. In the OOP+KT group, donor SD rats received 15 OOP treatments by transrectal insufflations (1 mg/kg), once a day, at an ozone concentration of 50 µg/ml, before the kidney transplantation. Twenty-four hours after transplantation, the parameters of renal function of the recipients were measured. The morphology and pathological effects of renal allograft were examined using hematoxylin and eosin staining, periodic acid-Schiff staining, a terminal deoxynucleotidyl transferase dUTP nick end labeling assay and immunohistochemistry. Markers of oxidative stress were also detected using the thiobarbituric acid method, and expression levels of Nrf-2 and HO-1 were determined by western blot analysis. Blood urea nitrogen and creatinine levels were significantly decreased in the OOP+KT group compared with the KT group, and the morphology and pathological changes of renal allograft were also less severe. Meanwhile, the renal allograft cell apoptosis index was significantly higher in the KT group compared to the OOP+KT group (P<0.05). Levels of superoxide dismutase, glutathione and catalase in the renal allografts were significantly higher in the OOP+KT group compared to those in the KT group (P<0.05), while malondialdehyde levels were significantly lower in the OOP+KT group compared to those in the KT group (P<0.05). Western blot analysis indicated that the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf-2) and heme oxygenase 1 (HO-1) were significantly higher in the OOP+KT compared to the KT group (P<0.05). In conclusion, the mechanism by which OOP alleviates oxidative stress injury in renal transplantation may be related to the activation of the signaling pathways of Nrf-2/HO-1 and inhibition of renal tubular epithelial cell apoptosis.

Thioredoxin-Interacting Protein Mediates NLRP3 Inflammasome Activation Involved in the Susceptibility to Ischemic Acute Kidney Injury in Diabetes.

Kidney in diabetic state is more sensitive to ischemic acute kidney injury (AKI). However, the underlying mechanisms remain unclear. Herein, we examined the impact of diabetes mellitus on thioredoxin-interacting protein (TXNIP) expression and whether mediated NLRP3 activation was associated with renal ischemia/reperfusion- (I/R-) induced AKI. In an in vivo model, streptozotocin-induced diabetic rats showed higher susceptibility to I/R injury with increased TXNIP expression, which was significantly attenuated by resveratrol (RES) treatment (10 mg/kg intraperitoneal daily injection for 7 consecutive days prior to I/R induction). RES treatment significantly inhibited TXNIP binding to NLRP3 in diabetic rats subjected to renal I/R injury. Furthermore, RES treatment significantly reduced cleaved caspase-1 expression and production of IL-1ß and IL-18. In an in vitro study using cultured human kidney proximal tubular cell (HK-2 cells) in high glucose condition (HG, 30 mM) subjected to hypoxia/reoxygenation (H/R), HG combined H/R (HH/R) stimulated TXNIP expression which was accompanied by increased NLRP3 expression, ROS generation, caspase-1 activity and IL-1ß levels, and aggravated HK-2 cells apoptosis. All these changes were significantly attenuated by TXNIP RNAi and RES treatment. In conclusion, our results demonstrate that TXNIP-mediated NLRP3 activation through oxidative stress is a key signaling mechanism in the susceptibility to AKI in diabetic models.

[The expression of GABA(A) receptor alpha1 and GABA(B) receptor 1 in medulla oblongata solitary nucleus and ambiguous nucleus in the cases of tramadol intoxication].

OBJECTIVE: To observe the expression of GABA(A) receptor alpha1 (GABA(A)alpha1) and GABA(B) receptor 1 (GABA(B)1) in human medulla oblongata solitary nucleus and ambiguous nucleus due to tramadol-induced death. METHODS: GABA(A)alpha1 and GABA(B)1 were detected by immunohistochemical SP method in tramadol-induced death group and control group. All results were evaluated by images analysis system. RESULTS: Low expression of GABA(A)alpha1 and GABA(B)1 were detected in solitary nucleus and ambiguous nucleus in the control brain tissue. In cases of tramadol-induced death, the expression of GABA(A)alpha1 and GABA(B)1 significantly increased. CONCLUSION: The mechanism of tramadol intoxication death could be caused by respiratory depression induced by over-expression of GABA(A)alpha1 and GABA(B)1 in medulla oblongata solitary nucleus and ambiguous nucleus.

Protective immunity induced in mice by multiantigenic DNA vaccine with genes encoding SAG1 and MIC8 of Toxoplasma gondii.

OBJECTIVE: To observe the immunoprotection induced by multiantigenic SAG1-MIC8 DNA vaccine of Toxoplasma gondii in C57BL/6J mice. METHODS: The sequences of genes encoding SAG1 and MIC8 protein were inserted into the eukaryotic expression vector pcDNA3.1 and the multiantigenic recombinant plasmid pcDNA3.1-SAG1-MIC8 was constructed. Then the recombinant plasmid was transfected into Hela cells to test its expression and the recombinant protein characterized by Western blotting 70 mice were divided into 5 groups randomly: PBS, pcDNA3.1, pcDNA3.1-SAG1, pcDNA3.1-MIC8 and pcDNA3.1-SAG1-MIC8. Each mouse was injected intra-muscularly by 100 microg recombinant plasmid for 3 times every two weeks. Mice were bled on day 0, 13, 27, 41, and 55. Four weeks after the final inoculation (on day 56), spleens from seven immunized mice per group were collected. Another seven immunized mice per group were intraperitoneally challenged with 1 x 10(4) tachyzoites of RH T. gondii and the survival time was observed. Serum IgG antibody and cytokines IFN-gamma and IL-4 were demonstrated by ELISA and the T lymphocyte proliferation assay were carried out with 3H-TdR incorporation. RESULTS: Western blotting showed that the mature protein extracts in Hela cells upon transfection with pcDNA3.1-SAG1 (Mr 34,000), pcDNA3.1-MIC8 (Mr 74,000) and pcDNA3.1-SAG1-MIC8 (Mr 109,000) were effectively expressed in cells. The results of IgG antibodies (on day 41 and 55), IgG2b, IgG2c, IFN-gamma (on day 55) and T lymphocyte proliferation assay (on day 56) were more obvious in mice immunized with pcDNA3.1-SAG1-MIC8 multiantigenic DNA vaccine than those in mice with single-gene plasmids (P < 0.05). There was no significant difference in IgG1 and IL-4 levels between vaccinated and control mice after the final inoculation (on day 55) (P > 0.05). The median survival time was 3, 4, 7, 7, and 10d, respectively, with considerable difference among the groups (P < 0.01). CONCLUSION: The multiantigenic DNA vaccine elicits a stronger immuno-protection in mice than the monovalent DNA vaccine.

[Tramadol intoxication and its toxicological effect].

Tramadol is a centrally acting analgesic with a dual mode of action. Its analgesic efficacy is attributed to its partial affinity for the mu-opiate receptor and its inhibition of norepinephrine and serotonin reuptake. Acting in a synergistic manner and being more efficacious, tramadol is used worldwide for the treatment of moderate to severe acute or chronic pain. Abuse and dependence of tramadol as well as tramadol-related deaths have been increasingly reported, either ingested alone or taken in combination with other potentially interacting drugs. The possible toxic effect of tramadol was reviewed from aspects of its analgesic mechanisms, adverse effect, dependence, and abuse.

Identify lymphatic metastasis-associated genes in mouse hepatocarcinoma cell lines using gene chip.

AIM: In order to obtain lymphogenous metastasis-associated genes, we compared the transcriptional profiles of mouse hepatocarcinoma cell lines Hca-F with highly lymphatic metastasis potential and Hca-P with low lymphatic metastasis potential. METHODS: Total RNA was isolated from Hca-F and Hca-P cells and synthesized into double-stranded cDNA. In vitro transcription double-stranded cDNA was labeled with biotin (i.e. biotin-labeled cRNA, used as the probe). The cRNA probes hybridized with Affymetrix GeneChip(r) MOE430A (containing 22 690 transcripts, including 14 500 known mouse genes and 4 371 ESTs) respectively and the signals were scanned by the GeneArray Scanner. The results were then analyzed by bioinformatics. RESULTS: Out of the 14 500 known genes investigated, 110 (0.8%) were up regulated at least 2(3) fold. Among the total 4 371 ESTs, 17 ESTs (0.4%) (data were not presented) were up regulated at least 2(3) fold. According to the Gene Ontology and TreeView analysis, the 110 genes were further classified into two groups: differential biological process profile and molecular function profile. CONCLUSION: Using high-throughput gene chip method, a large number of genes and their cellular functions about angiogenesis, cell adhesion, signal transduction, cell motility, transport, microtubule-based process, cytoskeleton organization and biogenesis, cell cycle, transcription, chaperone activity, motor activity, protein kinase activity, receptor binding and protein binding might be involved in the process of lymphatic metastasis and deserve to be used as potential candidates for further investigation. Cyclin D1, Fosl1, Hsp47, EGFR and AR, and Cav-1 are selected as the possible candidate genes of the metastatic phenotype, which need to be validated in later experiments. ESTs (data were not presented) might indicate novel genes associated with lymphatic metastasis. Validating the function of these genes is helpful to identify the key or candidate gene/pathway responsible for lymphatic metastasis, which might be used as the diagnostic markers and the therapeutic targets for lymphatic metastasis.

[An investigation on the transmission routes and early diagnosis of intrauterine infection induced by hepatitis B virus].

OBJECTIVES: To analyze the relationship between the fetus infection and HBV M, HBV DNA in amniotic fluid, umbilical cord blood, maternal blood and placenta, and to explore the mechanism of vertical transmission of HBV. METHODS: Immunonetric assay and nucleic acid amplification hybri-comb were used. Both HBV M and HBV DNA were detected in amniotic fluid, vein blood, umbilical cord blood for each of 65 HBV-positive women in their different gestational periods, while immunohistochemical analysis was carried out on the tissue of placenta, liver, lung or heart from each abortive fetus/dead infant in the case. RESULTS: For all of the 65 HBsAg-positive women in their different gestational periods, the detected positive rate of HBsAg was 21.50% in amniotic fluid, and 20.00% in umbilical blood. The positive rate of HBsAg, HBeAg, Anti-HBc and HBV DNA detected in blood, amniotic fluid and umbilical blood was 6.15%. The cases with positive HBsAg, Anti-HBe, Anti-HBc and negative HBV DNA were in a percentage of 13.85%. Immunohistochemical analysis on placentas after birth/abortion as well as the tissues of livers, lungs, hearts of the fetuses/dead infants in 4 cases of pregnant women with positive HBsAg, HBeAg, Anti-HBc or HBV DNA in blood, amniotic fluid or umbilical blood showed that HBsAg, HBcAg positive cells in the scope could be seen in every layer of the tissue of placenta, in the hepatic/pulmonary tissue, but not in the cardiac tissue. CONCLUSION: The infection in amniotic fluid or placenta relates to HBV infection in fetus; intrauterine HBV may result in infection in organs such as blood, liver, or lung of a fetus; infection in the amniotic fluid may be another key route of the intrauterine infection of fetus, and the detection on HBV M or HBV DNA in amniotic may be used as one of diagnostic proofs of HBV infection of fetus in its early stage.