Background and purpose:Silent information regulator proteins(sirtuins,SIRT)are a class Ⅲ histone deacetylases with nicotinamide adenine dinucleotide(NAD+)as coenzyme.YME1 like 1 ATPase(YME1L1)is essential for the maintenance of mitochondrial morphology,function and plasticity.Optic atrophy 1(OPA1)mainly mediates mitochondrial fusion.The aim of this study was to explore the expression of SIRT3 in the endocrine resistance of breast cancer,the relationship between SIRT3 and YME1L1 and OPA1,and the mechanism of SIRT3 in the endocrine resistance of breast cancer.Methods:4-hydroxytamoxifen was used to induce tamoxifen-resistant MCF-7/TAM cells.cell counting kit-8(CCK-8)was used to detect cell proliferation and verify drug resistance.The mitochondrial morphology was observed by transmission electron microscopy(TEM)and immunofluorescence staining.The expressions of SIRT3 and OPA1 were detected by real-time fluorescent quantitative polymerase chain reaction(RTFQ-PCR)and Western blot.JC-1 staining was used to detect mitochondrial membrane potential,and dihydroethidium(DHE)staining was used to detect reactive oxygen species(ROS)to verify mitochondrial function.SIRT3 was knocked down in drug-resistant cells by RNA interference,and SIRT3 and YME1L1 wild type(WT),simulated acetylation state mutant(MUT K237Q),and simulated deacetylation state mutant(MUT K237R)were overexpressed in parental cells by overexpression plasmid.Immunoprecipitation assay(IP)and immunofluorescence(IF)were used to verify the interaction between SIRT3 and YME1L1.Results:RTFQ-PCR and Western blot results showed that SIRT3 gene expression and protein level was significantly higher in drug-resistant cells than in parental cells.Overexpression of SIRT3 in parental cells decreased the sensitivity of breast cancer cells to tamoxifen.Knockdown of SIRT3 in drug-resistant cells enhanced the sensitivity of drug-resistant cells to tamoxifen.DHE staining showed that the ROS level was lower in tamoxifen resistant cells than in parental cells at the same concentration.Transmission electron microscopy and fluorescence staining showed that the mitochondria of the drug-resistant cells were elongated compared with the parental cells.Western blot results showed that the expression level of L-OPA1 protein was higher in drug-resistant cells than in parental cells.Overexpression of SIRT3 in the parental cells resulted in enhanced mitochondrial function and longer mitochondrial morphology compared with the control cells.Western blot showed that the expression of L-OPA1 was upregulated.When SIRT3 was knocked down in drug-resistant cells,the opposite result was obtained.We further verified how SIRT3 regulated OPA1 protein,affected the morphology and function of mitochondria,and promoted drug resistance of breast cancer.Overexpression of YME1L1(wild-type and mutant plasmids)in parental cells showed that overexpression of YME1L1 in the simulated deacetylation state resulted in similar results as overexpression of SIRT3,and overexpression of YME1L1 in the acetylated state resulted in similar results as knockdown of SIRT3.IP assay confirmed the interaction between SIRT3 and YME1L1 in breast cancer cells.The acetylation level of YME1L1 was different at different SIRT3 expression levels.IF assay showed that YME1L1 was co-localized with SIRT3 in MCF-7 cells.Conclusion:SIRT3 is highly expressed in tamoxifen-resistant breast cancer cells.SIRT3 upregulates L-OPA1 expression by deacetylating YME1L1,thereby promoting mitochondrial fusion and enhancing mitochondrial function,and promotes tamoxifen resistance in breast cancer.

Objective:To retrospectively analyze the viral levels and associated factors in patients with hepatitis B virus (HBV)-related primary liver cancer (PHC) in real-world settings and further explore the correlation between low viral load (LVL) and/or low-level viremia (LLV) and PHC.Methods:Five hundred twenty-four cases with HBV-related PHC with complete pathologically confirmed data from 2013 to 2020 were included. Percentages (%) were used to express their viral load, antiviral (oral) status, patient compliance, presence or absence of cirrhosis, family history of liver cancer, and others. LVL definition: After excluding detection errors by PCR method, serum HBV DNA <50-2 000 IU/ml, and those who had received antiviral drug treatment were called LLV. Antiviral treatment (AVT) rate definition: As of the confirmed diagnosis of PHC, those who had been regularly treated using oral antiviral drugs for six months or more (≥6 months).Results:General situation: The ratio of male to female enrolled patients was 15.90:1 (493/31). Patients aged >40 years accounted for 91.98% (482 cases). Hepatitis B surface antigen (HBsAg) positivity condition: The ratio of HBsAg-positive to HBsAg-negative/anti-HBc-positive (HBsAg-/anti-HBc+) PHC patients was 5.89:1 (448/76). Among the 76 HBsAg-/anti-HBc+patients, the ratio of HBsAg-/anti-HBs+/anti-HBc+ to HBsAg-/anti-HBs-/anti-HBc+ patients was 0.95:1 (37/39). Hepatitis B e antigen (HBeA) positivity condition: The ratio of HBeAg-negative to HBeAg-positive cases was 3.23:1 (400/124). HBV DNA level condition: The medical history records of 75.00% of patients (393/524) had traceable HBV DNA test reports. Out of 393 patients, 45.04% (177/393) accounted for undetectable HBV DNA, 13.49% (53/393) accounted for LVL, 41.48% (163/393) accounted for HBV DNA exceeding the upper limit of LVL, and 4.07% (16/393) accounted for LLV. Among HBsAg-positive and HBsAg-/anti-HBc+ patients, the HBV DNA positivity rates were 59.12% (214/362) and 6.45% (2/31), respectively. Antiviral treatment condition: Among the 448 HBsAg-positive PHC patients, the total AVT rate was 18.08% (81/448), of which seven patients did not have their HBV DNA results traced back. Among them, the AVT rate of 148 patients with HBV DNA lower than the lowest detection value was 41.22% (61/148); the AVT rate of 53 patients with LVL was 18.87% (10/53); and the AVT rate of 163 patients with HBV DNA≥LVL upper limit was 1.84% (3/163). Liver cirrhosis and family history condition: 348 patients (66.41%) had liver cirrhosis. 67 patients (12.79%) had a distinct family history of HBV-related liver cirrhosis and liver cancer. Alpha-fetoprotein (AFP) condition: 514 patients underwent AFP testing, with 30.93% of the patients had normal AFP levels, and 69.07% had AFP levels exceeding the upper limit of normal values (355/514). Among them, 10 μg/L400 μg/L accounted for 34.44% (177/514) and 34.63% (178/514), respectively. Tissue typing condition: 99.05% (519/524) were hepatocellular carcinoma, and well-differentiated and moderately differentiated cases accounted for only 8.59%.Conclusions:In the real world, comprehensive, timely screening, standardized, and effective antiviral treatment have not yet been achieved for patients with chronic hepatitis B, resulting in the persistence of HBsAg and/or HBV DNA positivity (especially LVL and/or LLV). Although LVL does not account for a high proportion among all HBV-related PHC populations, the vast majority of LVL populations have not received any antiviral treatment. In addition, some PHC populations with HBV DNA>2 000 IU/ml have not received standard antiviral treatment before the onset of the disease, which should be paid attention to. At the same time, HBsAg-/anti-HBc+ populations may have latent HBV infection and may even progress to PHC. Notably, PHC may still occur after HBeAg seronegative conversion.

New drug clinical trials have been considered as a positive way for treating cancer by cancer patients and doctors, and the extended dosing is a special way for patients' withdrawal from antitumor clinical trials to obtain investigational new drugs. However, neither the regulations of expanded dosing nor the detail documents for expanded dosing have been officially published in China. At present, expanded dosing of investigational drugs is still at the exploratory stage in various medical institutions, and a complete management system has not been established to meet patients' urgent needs for drug use. Based on the practical experience of extended dosing in Hunan Cancer Hospital, this paper preliminarily explored the application procedures and ethical review requirements of extended dosing for subjects in antitumor clinical trials. It is necessary to clarify the responsibilities of all patients in the procedure and establish a patient-medical institution-sponsor joint application system. In the process of ethical review, it is recommended that all parties fully consider the risks and benefits of extended dosing for patients, and then the ethics committee makes a comprehensive assessment to decide whether to approve extended dosing.
Humans ; China ; Physicians ; Antineoplastic Agents/therapeutic use*

OBJECTIVETo explore the effects of electrical stimulation at acupoints in the distribution area of auricular vagus nerve combined with sound masking on auditory brainstem response (ABR) and contents of neurotransmitters of γ-aminobutyric acid (γ-GABA), 5-hydroxytryptamine (5-HT) and acetyl choline (Ach) in inferior colliculus of tinnitus rats.

METHODSTwenty-four male adult SD rats were randomized into a control group, a model group, a 7-d treatment group and a 15-d treatment group. Except the control group, rats in the remaining groups were treated with intraperitoneal injection of 10% salicylate sodium at a dose of 350 mg/kg to establish tinnitus model. Rats in the control group were treated with injection of 0.9% NaCl. Rats in the 7-d treatment group and 15-d treatment group were treated with electrical stimulation at "Shenmen (TF₄)" and "Yidan (CO₁₁)" in the distribution area of auricular vagus nerve combined with sound masking, once a day, for 7 days and 15 days. The SigGenRP software of TDT system was applied to provide voice for single ear and collect the signal, and the voice threshold of ABR was tested. The levels of γ-GABA, 5-HT and Ach in inferior colliculus of rats were detected by enzyme linked immunosorbent assay (ELISA) and compared.

RESULTSCompared with the model group, the threshold values of ABR in 12 kHz and 16 kHz voice stimulation in the 7-d treatment group were significantly lower all P < 0.05); the threshold values of ABR from 4 kHz to 28 kHz voice stimulation in the 15-d treatment group were signally reduced (P < 0.05, P < 0.01), which was more significant than those in the 7-d treatment group. The level of γ-GABA in the model group was significantly lower than that in the control group (P < 0.05), and that in the 15-d treatment group was apparently higher than that in the model group (P < 0.05). The level of 5-HT in the model group was markedly higher than that in the control group (P < 0.05), and that in the 7-d treatment group was lower than that in the model group (P < 0.05), while that in the 15-d treatment group was apparently higher than that in the model group (P < 0.05). The level of Ach in the model group was obviously; lower than that in the control group (P < 0.05), and that in the 7-d treatment group was higher than that in the model group (P < 0.05).

CONCLUSIONElectrical stimulation at auricular vagus nerve combined with sound masking) could regulate the threshold of ABR, especially in the 15-d treatment group. This may be ascribed to modulating the levels of neurotransmitter of γ-GABA, 5-HT and Ach in inferior colliculus.

Acupuncture Points ; Animals ; Brain Stem ; physiopathology ; Electric Stimulation ; Evoked Potentials, Auditory, Brain Stem ; Humans ; Inferior Colliculi ; physiopathology ; Male ; Neurotransmitter Agents ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serotonin ; metabolism ; Tinnitus ; physiopathology ; therapy ; Vagus Nerve ; physiopathology ; gamma-Aminobutyric Acid ; metabolism

Objective To explore the clinical characteristics of patients with olfactory dysfunction after upper respiratory tract infection.Methods Through clinical specialist examination and imaging examination,95 cases of patients with olfactory dysfunction after upper respiratory tract infection were confirmed,58 cases in anosmia group and 37 cases in hyposmia group.All were performed by a subjective olfactometry (Sniffin'Sticks test) and a subjective taste function tests.The results were statistically analyzed by SPSS 17.0 software.Results In 58 cases of anosmia group,21 cases of male,37 cases of female;Twenty-six cases of youth,23 cases of middle age,9 cases of old age;Twenty-seven cases occurs in spring,11 cases in summer,12 in autumn and 8 in winter.Among 37 cases of hyposmia group,12 cases of male,25 cases of female;Eighteen cases of youth,16 cases of middle age,3 cases of old age;Fourteen cases occurs in spring,8 cases in summer,7 in autumn and 8 in winter.There was no statistically significant difference in gender,age and the onset season between the two groups (x2 =0.142,P ＞ 0.05;x2 =1.124,P ＞0.05;x2 =1.335,P ＞ 0.05).In anosmia group,with 4 cases of ageusia,22 cases of hypogeusia,32 cases of normal taste;in hyposrnia group,with 0 cases of ageusia,10 cases of hypogeusia,27 cases of normal taste.There were significant differences between the two groups with different types of taste disorder (Pearson correlation coefficient r =0.210,P ＜ 0.05),it was positive correlation.Conclusions It is suggested that after the upper respiratory tract infection,the olfactory dysfunction is often accompanied by the sense of taste dysfunction,the more severe the damage of olfactory function,the degree of damage to the taste function is also increased.Olfactory impairment degree exhibited no relationship with gender,age or onset seasons.

Objective To explore the rule of traditional Chinese medicine in treating tinnitus based on literature and Logistic multiple regression analysis. Methods Articles about tinnitus treatment using traditional Chinese medicine were searched in several databases, i.e. CNKI (1984-2013), VIP (1989-2013), CBM (1990-2013), and PubMed (1984-2013), and medication frequency was analyzed. Then, models for tinnitus medication were established through metrological method and Logistic multiple regression. Results The common prescriptions with highest frequency usage were Liuwei Dihuang Pill, Longdan Xiegan Decoction, and Erlong Zuoci Pill. Corni Fructus, Psoraleae Fructus, and Lycii Fructus were commonly used for kidney-deficiency type of tinnitus;Astragali Radix, Atractylodis Macrocephalae Rhizoma, and Codonopsis Radix were commonly used for the spleen-qi-deficiency type of tinnitus;Gardeniae Fructus, Gentianae Radix et Rhizoma, and Cyperi Rhizoma were commonly used for the liver-qi-dysfunction type of tinnitus;Pinelliae Rhizoma, Aurantii Fructus Immaturus, and Stephaniae Tetrandrae Radix were commonly used for the phlegm-fire disturbance type of tinnitus;Forsythiae Fructus, Menthae Haplocalycis Herba, and Mori Cortex were commonly used for the wind-fire-invasion type of tinnitus. Conclusion Analysis of medication frequency and Logistic multiple regression analysis can provide evidence and reference for the treatment of tinnitus and syndrome differentiation.

Objective To analyze the occurrence of clinical adverse drug reaction (ADR) of tonic Chinese medicinal herbs. Methods Articles about ADR of tonic Chinese medicinal herbs reported from January 1990 to February 2014 were collected from databases of CNKI, VIP, CBM, Pubmed and Embase according to incorporation and exclusion standard. Metrology analysis was also conducted. Results Totally 378 effective articles showed tonic Chinese medicinal herbs most likely to induce ADR were as follows:individual Chinese medicinal herbs including Polygoni Multiflori Radix, Ginseng Radix et Rhizoma, Lycii Fructus and Astragali Radix;injection containing Shenmai Injection, Shengmai Injection and Acanthopanax Injection;other herbal preparation covering Polygoni Multiflori tablets. The most common ADR involved the damage on skin tissue, nervous system and digestive system, with prominent relevant clinical symptoms such as itching, skin rash, dizziness, unconsciousness, nausea, vomiting, and liver dysfunction. Conclusion Correct understanding adverse reaction of tonic Chinese medicinal herbs will be conducive to rational drug use and avoiding risk.

OBJECTIVETo analyze the etiology, clinical features and prognosis of liver injuries caused by different drugs.

METHODSThe types of suspected drugs related to liver injury, clinical manifestations, liver biochemical parameters, clinical outcomes and other associated data were retrospectively assessed for 140 patients with drug-induced liver injury (DILI). The Roussel Uclaf Causality Assessment Method (RUCAM) was used to assess the causality between drugs and liver injury.

RESULTSThe most prevalent agents inducing DILI were Chinese traditional drugs (62.1%), followed by antipyretic analgesic drugs (10%) and antibiotics (5%). The ratio of male to female patients in the study cohort was 1:1.69, with 71 of the total patients (50.7%) being between the ages of 40 and 60 years-old. The RUCAM scale was not less than 3 points for any of the patients.In general, the clinical manifestations and biochemical results were not specific. The percentages of hepatocellular injury type, cholestatic injury type and mixed injury type were 51.4%, 30.7% and 17.9% respectively. The median age of patients with cholestatic liver injury was 55.6 years, which was older than that of patients with hepatocellular injury (47.1 years) or mixed injury (49.9 years).

CONCLUSIONAlthough antipyretic analgesics and antibiotics are considered as common drugs that can induce DILI, Chinese traditional drugs have emerged as another important group of liver injurious agents. Cholestatic DILI was found to occur more often in elderly patients than in younger patients.

Adult ; Anti-Bacterial Agents ; Chemical and Drug Induced Liver Injury ; Cholestasis ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Prevalence ; Prognosis ; Retrospective Studies

Objective:To evaluate the bioequivalence of two kinds of domestic nimesulide granules in healthy volunteers. Meth-ods:In self-control and two-way crossover design, 18 healthy male volunteers were divided into two groups randomly. Each subject was given 100 mg test or reference nimesulide granules with single dose. The concentration of nimesulide in plasma was determined by HPLC. The concentration of nimesulide in plasma was calculated and compared statistically to evaluate the bioequivalence between the two kinds of granules by DAS 2. 1 software. Results:The main pharmacokinetic parameters of test and reference preparations were as follows:Cmax was(9. 28 ± 2. 05) and(9. 41 ± 2. 31)μg·ml-1;Tmax was(3. 50 ± 1. 86)and(3. 56 ± 1. 65)h;T1/2 was (3. 43 ± 0. 85) and(3.38 ±0.68)h;AUC0-24 was(77.78 ±18.42)and(81.69 ±23.50)μg·ml·h-1;AUC(0-∞) was (79.07 ±19.21)and(82.92 ± 24. 11)μg·ml·h-1, respectively. The 90% confidential interval of ln(AUC0-24), ln(AUC0-∞) and ln(Cmax) of the test preparation was 90. 7%-107. 9%,90. 6%-111. 2% and 90. 7%-103. 0%, respectively. The relative bioavailability was (96. 7 ± 37. 6)%. Con-clusion:The two nimesulide granules are bioequivalent.

OBJECTIVE: To explore changes of olfactory and gustatory functions in patients with olfactory dysfunction. METHOD: The 284 study subjects included 92 healthy volunteers, 92 with hyposmia and 100 with functional anosmia. Their olfactory and gustatory functions were examined using T&T olfactometer, olfactory event-related potentials (OERPs) and triple drop method, respectively. RESULT: The T&T results showed that the difference between patients with hyposmia and functional anosmia and healthy subjects had statistical significance. The OERPs results showed that patients with olfactory dysfunction had N1 and P2 waves of prolonged latency and reduced amplitude when compared to healthy subjects with the difference of statistical significance. When compared to healthy subjects, patients with olfactory dysfunction had clear hypogeusia and the difference had statistical significance. There was no significant difference between female and male groups. CONCLUSION It is suggested that the apparently concomitant hypogeusia is in patients with olfactory dysfunction. Decreased of olfactory and gustatory function exhibited little or no relationship with gender. Combined examinations of OERPs, T&T and triple drop method, could make it possible to reflect the overall chemical sensory functions in subjects systematically and provide scientific evidence for clinical diagnosis and treatment early.
Adolescent ; Adult ; Aged ; Ageusia ; diagnosis ; physiopathology ; Case-Control Studies ; Evoked Potentials ; Female ; Humans ; Male ; Middle Aged ; Olfaction Disorders ; diagnosis ; physiopathology ; Smell ; Taste ; Young Adult