Mechanism of Polyphyllin Ⅱ in Induction of Ferroptosis in Hepatocellular Carcinoma HepG2 Cells / 中国实验方剂学杂志

ObjectiveTo investigate the induction of ferroptosis by polyphyllin Ⅱ （PPⅡ） in hepatocellular carcinoma HepG2 cells and its underlying mechanism. MethodThe effect of PPⅡ （0， 1.5， 3.0， 4.5， 6.0， 9.0， 18.0 mg·L-1） on the in vitro proliferation of HepG2 cells was assessed using the methyl thiazolyl tetrazolium （MTT） assay. Colony formation ability of HepG2 cells was evaluated through a colony formation assay. Cell migration ability was assessed via a scratch assay. Lactate dehydrogenase （LDH） content in HepG2 cells was measured using a kit. Reactive oxygen species （ROS） levels in HepG2 cells were observed using a fluorescence inverted microscope. Malondialdehyde （MDA）， glutathione （GSH）， and free Fe2+ content in HepG2 cells were detected using respective kits. The mitochondrial ultrastructure in HepG2 cells was observed by transmission electron microscopy. The expression of ferroptosis-related proteins p53， solute carrier family 7 member 11 （SLC7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， and transferrin receptor 1 （TFR1） in HepG2 cells was detected using Western blot. ResultCompared with the control group， the PPⅡ treatment groups showed significantly decreased survival rate of HepG2 cells in a dose-dependent manner （P<0.01）， significantly reduced number of cell colonies （P<0.01）， significantly shortened scratch healing distance， inverse correlation of the migration distance with drug concentration （P<0.01）， significantly increased LDH leakage in cells （P<0.01）， significantly enhanced relative fluorescence intensity of intracellular ROS， and significantly increased accumulation of lipid peroxide MDA （P<0.01）， decreased intracellular GSH content with increasing drug concentration （P<0.01）， and significantly enhanced fluorescence intensity of FeRhoNox-1 in cells （P<0.01）. Moreover， cells exhibited vacuolation， and mitochondria showed significant shrinkage with reduced or even disappeared cristae. Compared with the results in the control group， the expression of p53， ACSL4， and TFR1 proteins significantly increased， while the expression of SLC7A11 and GPX4 proteins significantly decreased in the PPⅡ treatment groups （P<0.05）. ConclusionIn summary， PPⅡ induces ferroptosis in HepG2 cells by regulating the p53/SLC7A11/GPX4 signaling axis， promoting ACSL4 expression and Fe3+ uptake， leading to an imbalance in the antioxidant system.

Active Ingredients of Chinese Medicines Induce Ferroptosis in Tumor Cells： A Review / 中国实验方剂学杂志

Ferroptosis， a new form of programmed cell death different from apoptosis， necrosis， and autophagy， is closely associated with a variety of physiological and pathological processes. Iron-mediated accumulation of reactive oxygen species is the main inducement of ferroptosis， the mechanism of which is related to intracellular lipid metabolism， iron metabolism， and antioxidant defense pathways. Multiple signaling axes and regulators jointly regulate the occurrence and disruption of ferroptosis. Studies have demonstrated that ferroptosis regulates the growth and proliferation of tumor cells. Inducing ferroptosis in tumor cells can control the growth， metastasis， and multi-drug resistance of tumors. Therefore， the effect and mechanism of ferroptosis on tumor cells have become a hot topic in anti-cancer research. As the research advances， a variety of ferroptosis inducers has been used in the clinical chemotherapy for cancers and demonstrate significant efficacy. Accordingly， the development of ferroptosis-inducing anticancer drugs has become a new research direction for tumor treatment. Some active ingredients such as lycorine， oleanolic acid， dihydroartemisinin， pseudolaric acid B， and ophiopogonin B of Chinese medicines can induce ferroptosis in tumor cells via lipid metabolism， iron metabolism， system Xc-， and GPX4/GSH to regulate the development of tumors， demonstrating a promising prospect in clinical treatment. Based on the theory of the mechanism of ferroptosis， this paper reviews the research progress in ferroptosis induced by active ingredients of Chinese medicines in tumor cells and describes the metabolic regulatory network of ferroptosis from signaling pathways and regulatory factors， providing new strategies for applying active ingredients of Chinese medicines in the treatment of tumors.

Total Saponins in Paridis Rhizoma： A Review / 中国实验方剂学杂志

Paridis Rhizoma possesses the functions of clearing heat and detoxifying， alleviating swelling and relieving pain， cooling the liver and calming the convulsion. Saponins are the main active components of Paridis Rhizoma. Studies have shown that total saponins in Paridis Rhizoma have obvious inhibitory effect on solid tumors such as breast cancer， lung cancer， gastric cancer， and liver cancer and non-solid tumors such as leukemia. The saponins may exert the anti-tumor effects by inhibiting the proliferation， migration， and invasion of tumor cells， regulating cell cycle， inducing apoptotic and non-apoptotic death pathways， and regulating metabolism and tumor microenvironment. Furthermore， total saponins in Paridis Rhizoma showed anti-inflammatory， antioxidant， antimicrobial， hemostatic， and uterus-contracting activities. At the same time， they may induce apoptosis of normal cells， inflammation and oxidative stress， and metabolic disorders. In recent years， the reports of liver injury， reproductive injury， gastrointestinal injury， hemolysis， and other adverse reactions caused by total saponins in Paridis Rhizoma have been increasing. Pharmacokinetic studies have shown that there are significant differences in the metabolism of total saponins in Paridis Rhizoma administrated in different ways. Injection has a fast clearance rate， while oral administration may have hepatoenteric circulation. Meanwhile， due to the low solubility and activation of P-glycoprotein （P-gp） molecular pump， the prototype absorption， intestinal permeability， and recovery rate of total saponins in Paridis Rhizoma are poor， which affects the bioavailability. The bioavailability can be improved to some extent by preparing new dosage forms or new drug delivery systems with advanced technology. This paper reviews the pharmacological effect， pharmacokinetics， and adverse reactions of Rhizoma Paridis total saponins by searching the China National Knowledge Infrastructure （CNKI）， VIP， and Web of Science with ''Rhizoma Paridis total saponins'' as the keywords， hoping to provide references for the research， development， and clinical application of such components.

A case report of middle ear cholesteatoma complicated with labyrinthine fistulaand delayed endolymphatic hydrops / 临床耳鼻咽喉头颈外科杂志

Delayed endolymphatic hydrops （DEH） is a rare disease that causes vertigo and is often misdiagnosed as other vertigo diseases. This article reports on a patient with vertigo who was easily misdiagnosed. The patient was a middle ear cholesteatoma complicated by labyrinthine fistula （LF）; however, his vertigo was episodic vertigo, which could not be explained solely by LF causing labyrinthitis. The possibility of endolymphatic hydrops was suspected, which was confirmed by inner ear magnetic resonance gadolinium imaging. This is the first reported case of middle ear cholesteatoma complicated by LF and DEH. The patient underwent surgical resection of the cholesteatoma and three semicircular canal obstructions at the same time. During two years postoperative follow-up, the patient did not experience a recurrence of vertigo. When diagnosing vertigo diseases, a careful history of vertigo is of utmost importance.

Application of a new double-headed endoscopic cleaning brush to soft endoscope cleaning / 中华消化内镜杂志

In order to clean the endoscopic tube more effectively, a new double-headed endoscopic cleaning brush was used in this study. A total of 130 colonoscopies were selected from the Digestive Endoscopy Center of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine from June 2019 to August 2020. The colonoscopy cleaning sequence was marked with odd and even number. Colonoscopies marked with the odd number were assigned to the conventional group ( n=65) which received back and forth cleaning with the single-headed endoscope cleaning brush. Colonoscopies marked with the even number were assigned to the experimental group ( n=65) which received one-way cleaning with a nylon brush at the head and a dense non-woven brush at the tail. The cleaning methods for endoscopes were in accordance with Flexible Endoscopic Cleaning and Disinfection Technical Specification WS507—2016. ATP bioluminescence tests and bacterial quantitative cultures were applied to evaluate the cleaning effects of the two methods. The results showed that ATP relative light unit (RLU) decreased in both the experimental group and the conventional group [530.63 RLU (26-3 559 RLU) VS 270.87 RLU (20-1 415 RLU)] before and after cleaning the endoscope tube, showing significant difference between the two groups ( Z=-2.894, P<0.05). After scrubbing, the positive rate of bacterial culture on the brush head was 86.2% (56/65) for the double-headed non-woven brush head, 46.2% (30/65) for the double-headed nylon brush head and 32.3% (21/65) for the single-headed nylon brush head with significant differences among the three groups ( χ 2=41.046, P<0.05). The cleaning effect of the new double-headed endoscopic cleaning brush is better than that of the conventional single-headed endoscopic cleaning brush for soft endoscope cleaning. Non-woven brush is better than nylon brush in eliminating bacteria.

The Functions of the Middle Ears of Children with Adenoid Hypertrophy / 听力学及言语疾病杂志

Objective To explore the influence of adenoid hypertrophy on the state of the middle ear in children.Methods Two hundred and seventy two adenoid hypertrophy patients,aged from 2 to 12 years old,were examined with otoscopy,tympanometry and temporal bone computered tomography before adenoidectomy.The average age of the patients was 6.3 years old.These patients were divided into two groups:Group A contained 94 normal hearing patients(188 ears),Group B 178 patients(356 ears) containing of hearing loss.Results Out of the temporal bone computed tomography (CT) revealed that 209 patients(396 ears) had middle ear effusions in(72.79%,396/544ears),and 37 patients(65/188 ears,34.57%) hydrotympanum in Group A and 172 patients(331/356 ears,92.98%) in Group B.All the 396 ears effusion were confirmed by operation.Three hundred seventy three ears effusion were confirmed by the CT and operation in the two groups,377 ears with B type tympanogram(373/377,98.94%),93.85%(61/65 ears) in Group A,100%(312/312 ears) in Group B,respectively.The positive predictive value for middle ear effusion of the B-type tympanogram was higher in Group B than in Group A(P＜0.01).Fourteen ears with peak pressure ＜-200 daPa revealed hydrotyrnpanum and 23 ears with normal acoustic stapedius reflex revealed no fluid by CT and operation in 73 ears with C-type tympanogram.CT also revealed two patients suffered from large vestibular aqueduct syndrome and one with cochlear malformation in Group B.Conclusion Type B tracing tympanogram has a high positive predictive value for middle ear effusion in adenoid hypertrophy children.Type C tympanogram could not exclude effusion when the peak pressure is＜-200 daPa.CT is the best tool for identifying hydrotympanum.

Diagnosis and management of pulsatile tinnitus of venous origin / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To discuss the diagnosis and management of pulsatile tinnitus of venous origin.@*METHOD@#A retrospective study was conducted on 12 patients who were diagnosed with pulsatile tinnitus of venous origin and treated with ligation of internal jugular veins. We reevaluated the evidences of identifying pulsatile tinnitus of venous origin and reviewed the short-term and long-term postoperative effects and complications. We also reviewed associated articles in this report.@*RESULT@#Seven patients got relief of tinnitus in less than one week after the surgery, while the other 5 patients had no relief. Seven patients were inquired in this study and the other five lost to follow-up. According to the long review (from one to five years postoperatively), two patients who acquired immediate effect got relief of tinnitus, four including complained of no relief and the seventh aggravated into roaring. Three patients who got no immediate relief got no improvement at all. No one in our review complained of any complications.@*CONCLUSION@#It's assumed that a history of pulsatile tinnitus, alleviation of tinnitus when pressing jugular veins, tinnitus changing with head position or posture and no occupying lesion in temporal CT scan or cranial MRI are inadequate in diagnosing pulsatile tinnitus of venous origin. Vascular imaging is also necessary to exclude other pathological changes like dura arteriovenous fistula, sigmoid diverticulum and so on. CT arteriography and venography are recommended preferentially. Ligation of internal jugular veins is controversial in patients who have no absence of transverse and sigmoid sinus and identified as pulsatile tinnitus of venous origin.

The epidemiological study of methicillin-resistant Staphylococcus aureus in Shanghai / 中华检验医学杂志

Objective To investigate antibiotic resistance and molecular epidemiology profile of methicillin-resistant Staphylococcus aureus (MRSA) in Shanghai. Methods The antibiograms of 140 MRSA isolates from 5 hospitals for 13 drugs were analyzed by agar dilution and broth dilution. The PVL gene and SCCmec were detected by PCR; The clonal relatedness of 140 isolates were determined by PFGE and 39 strains were chosen to be characterized further by spa typing. Results All 140 MRSA are PVL negative and most of them were identified as SCCmec Ⅲ [45.7% (64/140)], followed by SCCmec Ⅲ a [25.0% (35/140)], SCCmecⅢb [14.3% (20/140)], SCCmecⅡ [10.7% (15/140)] and SCCmecⅣ [4.3% (6/140)]. All isolates were susceptible to vancomycin, teicoplanin and daptomycin. The resistance to gentamicin, sulphamethoxazole and clindamycin was 98. 6% (138/140), 98. 6% (137/140) and 97. 9% (137/140), respectively. Resistance to erythromycin, ciprofloxacin and tetracycline was above 80%, and resistance to rifampicin was 10. 7% (15/140). Sixteen different PFGE patterns(A-P) were found and most of MRSA belonged to group C[30. 7% (43/140)] ,B[13.6% (19/140)]and Ⅰ [10. 7% (15/140)]. Among 39 strains with prevalent PFGE patterns, 4 spa genotypes were identified: t002133. 33% (13/39)] ,t030 [12. 82% (5/39)] ,t037[51.28% (20/39)]and t459[2. 57% (1/39)]. Conclusions Sixteen different PFGE patterns and 4 spa genotypos were found from 5 hospitals in Shanghai. The most popular MRSA clone is PVL negative, SCCmec Ⅲ, with resistant profile of erythromycin, ciprofloxacin,clindamycin,etracycline, gentamicin,and sulphamethoxazole [E-C-L-T-G-M-]. This result suggests that hospital infection control and reasonable antibiotic usage are critical.

Leukocidin prevalence and molecular epidemiology of methicillin-resistant Staphy-lococcus aureus / 中国感染与化疗杂志

Objective Methicillin-resistant Staphylococcus aureus (MRSA) is a growing public health concern that has been associated with pediatric fatalities. This study investigated the genotypes of staphylococcal cassette chromosomal mec (SCCmec) and Panton-Valentine Leukocidin (PVL) in MRSA strains isolated from Shanghai Children's Hospital by PCR. Methods A total of 30 strains of MRSA were isolated from various clinical specimens from October 2005 to June 2006. The antimicrobial susceptibility was measured by agar diffusion method. SCCmec typing was conducted using a novel multiplex PCR assay allowing for concomitant detection of methicillin resistance (mecA gene) to facilitate detection and classification of all currently described SCCmec typesⅠ, Ⅱ, Ⅲ, Ⅳa, b, c, d andⅤ. PVL gene was also determined by PCR. Results mecA gene was positive in all the strains. SCCmecⅡ was identified in 6(20.0%) isolates, SCCmecⅢ in 15(50.0%) isolates, SCCmecⅤ in 2 and SCCmecⅣa in 1 isolate. Six MRSA strains were non-typeable. The isolates with SCCmecⅡ or SCCmecⅢ were resistant to multiple antibiotics. The strains harboring SCCmecⅣa or SCCmecⅤwere susceptible to all antibiotics except β-lactams. Eleven (36.7%) isolates were PVL positive. The genotypes and subgenotypes of staphylococcal chromosomal cassette mec of eleven PVL-positive MRSA were SCCmecⅡ(1 isolates), SCCmecⅢ (5 isolates), SCCmecⅣa (1 isolate), SCCmecⅤ (2 iso-Lates) non-typeable (2 isolates). Conclusions SCCmecⅡ and SCCmecⅢ are the major genotypes of MRSA in our hospital. These isolates are multi-resistant to antibiotics. The prevalence of PVL gene is higher in SCCmecⅡ- or SCCmeⅢ-positive MRSA. The isolates with SCCmecⅡ or SCCmecⅢ were resistant to multiple antibiotics.

Antimicrobial Resistance and Staphylococcal Cassette Chromosome mec Genotype of Meticillin-resistant Staphylococcus aureus from Lower Respiratory Tract / 中华医院感染学杂志

OBJECTIVE To investigate antimicrobial resistance and molecular epidemiology profiles of meticillin-resistant Staphylococcus aureus(MRSA)sampled from lower respiratory tract.METHODS Totally 107 MRSA strains were isolated from lower respiratory tract specimens at Shanghai Pulmonary Hospital between Dec 2005 and Dec 2006.PVL genes were detected by PCR.The genotypes of SCCmec were identified by multiplex PCR.The antimicrobial resistance of MRSA were tested by Kirby-Bauer agar dilution.We also performed the homology of 32 MRSA strains using pulsed-field gel electrophoresis(PFGE).RESULTS All of the 107 MRSA strains were negative in the PVL locus detection and the most frequent SCCmec types were type Ⅲ(81.3%),the others including type Ⅱ(15.9%),type Ⅳ(2.8%),type Ⅰ and type Ⅴ were not found in this group.Those 3 different types of SCCmec were all resistant to ?-lactam antibiotics,less resistant to rifampin,and susceptible to vancomycin,teicoplanin and daptomycin.The resistant rate of those 3 types were different to the non-?-lactam antimicrobial drugs such as trimethoprim/sulfamethoxazole,clindamycin,erythromycin,gentamicin,levofloxacin,and tetracycline,the resistant rate in the types Ⅱ and Ⅲ was significantly higher than the type Ⅳ.PFGE analyses assorted the 32 MRSA strains into 4 PFGE patterns:pulsotype A(25 strains),including subtypes A1(17strains),A2(1 strain)and A3(7 strains);pulsotype B(5 strains),pulsotype C(1 strain),and pulsotype D(1 strain).CONCLUSIONS This study does not found positive PVL locus in the MRSA strains in our hospital,the most frequent SCCmec types are type Ⅲ and some are type Ⅱ.PFGE presented that there are outbreaks of MRSA in ICU ward and TB ward No 5 at that time and the pandemic strains are subtypes A1 and A3,most of these MRSA strains are multiple resistant,which deserves attention from both the clinical staff and infection-control department of the hospital.