ObjectiveTo explore the potential mechanism of Zuoguiwan in ameliorating polycystic ovary syndrome （PCOS） by network pharmacology and liquid chromatography-mass spectrometry （LC-MS）-based metabolomics. MethodThe active ingredients and potential targets of Zuoguiwan for treating PCOS were predicted by bioinformatics. SD rats were assigned into a control group and a modeling group. The rat model of PCOS was established by gavage with letrozole （1 mg·kg^-1） combined with feeding with a high-fat diet. At the end of modeling， the modeled rats were assigned into model （normal saline）， metformin （300 mg·kg^-1）， and Zuoguiwan （concentrate 1.62 g·kg^-1） groups. The body weight and oestrous cycle of each rat were recorded， and the ovary was stained with hematoxylin and eosin for observation of ovarian morphology. Enzyme-linked immunosorbent assay was employed to determine the serum levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， anti-mullerian hormone （AMH）， testosterone （T）， and estradiol （E₂）， and the LH/FSH ratio was calculated. Serum metabolomics of rats was conducted by orthogonal partial least squares-discriminant analysis （OPLS-DA） to screen the metabolite-enriched pathways. Furthermore， network pharmacology and association analysis were employed construct the compound-response-enzyme-gene network. ResultA total of 503 potential targets of Zuoguiwan and 5 843 targets of PCOS were screened out， with 271 common targets. The Gene Ontology enrichment analysis revealed that the common targets were involved in the response to lipopolysaccharide， etc.， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment yielded 119 pathways. Animal experiments showed that compared with the control group， the model group presented increased body weight （P<0.01）， elevated LH and AMH levels （P<0.01）， increased LH/FSH ratio （P<0.01）， lowered E₂ level （P<0.01）， and increased cystic follicles. Compared with the model group， Zuoguiwan and metformin decreased the body weight （P<0.01）， reduced atretic follicles and cystic follicles， increased mature follicles and corpus luteum， and thickened the granulosa layer. Moreover， Zuoguiwan lowered the T， FSH， LH， and AMH， and LH/FSH levels （P<0.01） and elevated the E₂ level （P<0.01）. The principal component analysis and OPLS-DA in metabolomics showed that the differential metabolites between Zuoguiwan and model groups included 26 up-regulated metabolites in the Zuoguiwan group. There were 8 common pathways predicted by the KEGG enrichment analysis in network pharmacology and the metabolite enrichment in metabolomics. The results of topological analysis revealed the pathways of steroid hormone biosynthesis and glycerol-phospholipid metabolism， and the constructed compound-response-enzyme-gene network revealed that the key targets were protein kinase B1 （Akt1）， epithelial growth factor receptor （EGFR）， prostaglandin-endoperoxide synthase 2 （PTGS2）， and mitogen-activated protein kinase 1 （MAPK1）. ConclusionZuoguiwan regulated the steroid hormone biosynthesis pathway to recover hormone levels， promote follicle production and development， and improve ovarian function， which may be the potential mechanism of this medicine in treating PCOS.

Objective:To investigate the effect of gastrodin(GAS)on the sex-determining region Y-box2(SOX2)/β-catenin pathway in microglia induced by lipopolysaccharide(LPS).Methods:BV2 microglia was cultured in vitro and divided into the following groups:Control group(Control),LPS group(LPS),LPS+0.17 mmol/L gastrodin treatment group(LPS+GAS-L),LPS+0.34 mmol/L gastrodin treatment group(LPS+GAS-H),SOX2 inhibitor pronethalolgroup(PR),LPS+PR group(LPS+PR),and LPS+PR+GAS group(LPS+PR+GAS).Effect of PR on BV2 microglia viability was detected by CCK-8.The expression of SOX2,β-catenin,mannose receptor(CD206)and tumor necrosis factor-α(TNF-α)was assessed using Western Blot and immunofluorescence double staining.Results:PR did not induce significant BV2 cell death in the 0～40 μmol/L range.After LPS treatment,the expression levels of SOX2,β-catenin,and TNF-α significantly increased in the LPS group,while CD206 decreased(P＜0.05).Following GAS treatment,the expression levels of SOX2,β-catenin,and TNF-α significantly decreased,while CD206 increased(P＜0.05).Compared to the LPS group,the expression levels of β-catenin and TNF-α significantly de-creased in the PR group(P＜0.05),but no significant difference was observed between the LPS+GAS and LPS+PR+GAS group.Conclusion:GAS significantly inhibits LPS-induced microglia activation potentially through the inhibi-tion of the SOX2/β-catenin signaling pathway,and exerts anti-inflammatory effects.

Objective:To analyze the clinical and epidemiological features of human rhinovirus (HRV) infection in adult patients with upper respiratory tract infection (URTI) in Nanjing.Methods:Epidemiological data of adult patients with URTI in Nanjing from October 2021 to September 2022 were collected. Clinical specimens were collected and subjected to quantitative reverse transcription polymerase chain reaction (qRT-PCR) for the detection of 14 common respiratory viruses. The VP4/VP2 genes in HRV-positive samples were amplified and sequenced. Then a phylogenetic tree was constructed.Results:A total of 399 pharyngeal swabs were collected from patients with URTI. The overall positive rate of respiratory viruses was 28.07% (112/399) with HRV accounting for most at 9.52% (38/399). Thirty-seven VP4/VP2 sequences were successfully obtained from the 38 HRV-positive specimens. Three genotypes involving 25 serotypes were identified with 13 strains belonging to HRV-A, 14 belonging to HRV-B, and 10 belonging to HRV-C. The three genotypes of HRV showed alternate prevalence or co-prevalence.Conclusions:HRV was the main pathogen causing URTI in adult patients in Nanjing from October 2021 to September 2022, and three genotypes of HRV-A, B and C were prevalent alternatively or together.

Objective:To explore the correlation between the single nucleotide variation of profibrin-1 (PFN1) gene and secondary osteoporosis (OP) after stroke and its influence on bone metabolism indexes.Methods:120 patients with post-stroke hemiplegia who were treated in our hospital from Jan. 2019 to Jun. 2023 were selected as study objects and divided into OP group and non-OP group. Levels of vitamin D[25- (OH) D], tartrate-resistant acid phosphatase (TRAP) , osteocalcin (BGP) , serum type I procollagen amino terminal prolongation brain (P1NP) and type I collagen basal terminal β special sequence (β-CTX) were detected in all patients. Two SNPS (rs6559 and rs78224458) in PFN1 gene were genotyped.Results:There were significant differences in serum 25- (OH) D, TRAP, P1NP and β-CTX levels between OP group and non-OP group ( P<0.05) . The GG, GA and AA genotypes at rs6559 of PFN1 gene were significantly different between OP and non-OP patients ( P<0.05) . The combined model showed that compared with GG genotype carriers, the risk of secondary OP in GA and AA genotype carriers was 3.250 and 5.417 times higher, respectively. The results of the dominant model showed that the risk of secondary OP was 3.792 times higher in patients with mutant genes (GA or AA) than in patients with GG genotype. Recessive model results showed that patients with AA genotype had a 3.810-fold increased risk of secondary OP compared with GG and GA carriers. There was no significant difference in TT, TC, CC genotype distribution, genetic model and allele frequency at rs78224458 of PFN1 gene between OP patients and non-OP patients ( P>0.05) . There were no significant differences in 25- (OH) D, TRAP or BGP among the rs6559 GG, GA and AA genotypes of PFN1 gene ( P>0.05) , while there were significant differences in P1NP andβ-CTX levels among the three groups ( P<0.05) . Conclusion:The rs78224458 variation of PFN1 gene is associated with secondary OP in patients with hemiplegia after stroke, and may affect the bone metabolism indexes of patients.

AIM: To investigate the effect of oxypeucedanin (OPD) on doxorubicin resistance in human breast cancer MCF-7/DOX cells and its possible mechanism. METHODS: MCF-7/DOX cells were cultured in vitro, MTT assay was used to detect the effect of OPD on the survival of MCF-7/DOX cells, and the effect of OPD combined with different concentrations of doxorubicin on the proliferation of MCF-7/DOX cells were investigated. The effect of OPD combined with doxorubicin on the expression of genes including MDR1, MRP1, AGPAT2, CHKA, CEPT1, DGKA, PCYT1A, PLA2G15 in MCF-7/DOX cells was measured by qRT-PCR. The effect of OPD combined with doxorubicin on the protein expression of MDR1, MRP1, CHKA and CCTα in MCF-7/DOX cells was determined by Western blot. RESULTS: The IC

Objective To explore the relationship between asymmetrically hypointense veins (AHVs) on susceptibility-weighted imaging (SWI) and collateral circulation.Methods We retrospectively enrolled acute ischemic stroke patients with severe stenosis or occlusion of M1 segment of the middle cerebral artery ± intracranial internal carotid artery.All the patients underwent diffusion-weighted imaging (DWI),SWI,and computed tomography angiography (CTA) of intracranial and cervical arteries within 72 hours from symptom onset.We explored the association of the level of AHVs with the degree of the regional leptomeningeal score (rLMC) on baseline CTA and other clinical and image data.The factors that might influence the prognosis of stroke were also analyzed.Results Fifteen patients with mild AHVs and 15 with extensive AHVs were enrolled in our study.The level of AHVs was positively correlated with CTA rLMC (r =0.481,P =0.007) and the degree of collateral circulation (r =0.402,P =0.028).Patients with extensive AHVs had better collateral status,smaller DWI infarction lesion ((11.62 ± 9.07) ml vs (95.77 ± 91.12) ml,t =3.559,P =0.001),and lower NIHSS scores on admission (6.47 ± 4.34 vs 12.33 ± 7.60,t =2.595,P =0.015)and at discharge (4.80 ± 4.69 vs 9.60 ± 7.03,t =2.200,P =0.036).The high degree of rLMC,small DWI lesion,young age and lower NIHSS scores,but not extensive AHVs were related with favorable outcome at 3 months after stroke.Conclusion Extensive AHVs can reflect good collateral circulation to some extent,but cannot be equivalent to or replace the collateral status.

Objective To explore the effectiveness and compliance of ketogenic-diet(KD) treatment for infantile spasm(IS).Methods Ninety-eight IS patients who were treated with KD in Wuhan Children's Hospital from March 2009 to June 2015 were analyzed by using retrospective case-control study,the patients were divided into 4 groups:newly diagnosed IS patients group (group A,including 24 patients),one antiepileptic drug (AEDs) failure IS patients (group B,including 28 patients),two and more AEDs failure IS patients (group C,including 29 patients),and two or more AEDs combined with ACTH failure IS patients(group D,including 17 patients).The spasm-free andretention rates after 3,6 and 12 months KD treatment were compared among these groups.Results Overall retention rate was 80.6％ (79/98 cases),69.4％ (68/98 cases),and 42.9％ (42/98 cases)at 3,6,12 months,respectively.The 3-month retention rate in group A,B,C and D was 83.3 ％ (20/24 cases),78.6％ (22/28 cases),82.7％ (24/29 cases) and 76.4％ (13/17 cases) respectively,and there was no significant difference among these groups (P ＞ 0.05).The 6-month retention rates in each group was 75.0％ (18/24 cases),67.9％ (19/28 cases),68.8％ (20/29 cases) and 65.0％ (11/17 cases) in sequence,and there was also no significant difference among these groups(P ＞0.05).The 12-month retention rate was 54.2％ (13/24 cases),21.4％ (6/28 cases),48.3％ (14/29 cases) and 52.9％ (9/17 cases) in group A,B,C and D in sequence,the 12-month retention rate of group B was significantly lower than that of other 3 groups,and the differences were statistically significant(x2 =5.973,4.508,4.727,all P ＜ 0.05),and there was no significant difference among the A,C,D groups (all P ＞ 0.05).The spasm-free rate at 3,6,12 months of KD treatment was 19.4％ (19/98 cases),20.4％ (20/98 cases),30.6％ (30/98 cases).The 3-month spasm-free rate in A,B,C,D groups were as follow:41.7％ (10/24 cases),14.3％ (4/28 cases),10.3％ (3/29 cases),11.8％ (2/17 cases),respectively.The 3-month spasm-free rate in group A was significantly higher than that of other 3 groups,and the differences were statistically significant (x2 =10.238,9.219,6.697,all P ＜ 0.05),but there was no significant difference among the B,C,D groups (all P ＞ 0.05).The 6-month spasm-free rates were 41.7％ (10/24 cases),14.3％ (4/28 cases),13.8％ (4/29 cases),and 11.8％ (2/17 cases) in group A,B,C and D in order,and the spasm-free rate in group A was significantly higher than that of other 3 groups,and the differences were statistically significant(x2 =4.924,5.249,4.298,all P ＜ 0.05),but there was no significant difference among the A,C,D groups (all P ＞ 0.05).The 12-month spasm-free rates were 54.2％ (13/24 cases),21.4％ (6/28 cases),24.1％ (7/29 cases),and 23.5 ％ (4/17 cases) in group A,B,C and D,and the spasm-free rate in group A was significantly higher than that in other 3 groups,and the differences were statistically significant(x2 =8.354,7.923,4.364,all P ＜ 0.05),but there was no significant difference among the A,C,D groups (all P ＞ 0.05).Conclusions The spasm-free rate of KD therapy for newly-diagnosed IS is higher than that of IS patients whose drug-therapy failed.KD therapy may be the top priority for IS patients and part of those patients whose drug-therapy failed can still get seizure-free with KD diet.

Objective To investigate the relationship of the cytogenetic abnormalities detected by FISH in patients with MM and their clinical features. Methods FISH on bone marrow (BM) cells was performed in 57 enrolled MM patients. Relationships between cytogenetic abnormalities and clinical features were analyzed. Results By statistical analysis , both D13S319 deletion and RB1 deletion were associated with high level of serum LDH (P = 0.024; P = 0.018) and BM plasma cells index (P = 0.027; P = 0.013). 1q21 amplification was significantly associated with high level of LDH (P = 0.030 ) and the occurence of light chain type myeloma (P = 0.023). IgH rearrangement was associated with renal function damage (P = 0.009). There were correlations among D13S319 deletion, RB1 deletion, 1q21 amplification and IgH rearrangement (P<0.01). Conclusion The genetic abnormalities detected by FISH in patients with MM were correlated with various clinical poor prognostic indicators, which can evaluate the condition and prognosis of patients more efficiently.

OBJECTIVETo investigate the clinical and imaging characteristics of chronic otitis media with intact tympanic membrane.

METHODTen patients were retrospective studied in the department of otorhinolaryngology of Guangdong general hospital from December 2006 to January 2015. The clinical characteristics of their manifestations, audiology and imaging examinations were analyzed.

RESULTSAll the cases could be divided into two groups: the intracranial complication group who was primarily diagnosed as intracranial complications, and the hearing loss group who primarily complained of hearing loss. Five cases belonged to the first group, which include 1 cerebellar abscess, 3 meningitis and 1 meningoencephalitis, 2 of them were profound hearing loss, 2 were mixed hearing loss, and 1 was normal hearing. The other 5 cases belonged to the hearing loss group, 3 were mixed deafness, 2 were conductive deafness. All the case showed positive high-resolution computed tomography (HRCT) features. In the first group, four cases showed the soft tissue shadow in tympanic antrum/superior tympanic pouch and 1 case showed a wide damage of skull base, in addition to intracranial complications. In the second group, five cases showed soft tissue imaging in tympanic antrum/superior tympanic pouch. Pathology showed that 2 cases were cholesterol granuloma(one was in the first group and the other was in the second group), 4 were intracranial inflammatory(the first group) and 4 were cholesteatoma(the seond group).

CONCLUSIONSIn chronic otitis media with intact tympanic membrane, intracranial complications may be the primarily diagnosis, which should be paid much attention by multidisciplinary team. HRCT and audiology were valuable for early diagnosis.

Brain Abscess ; etiology ; Cerebellar Diseases ; etiology ; Cholesteatoma ; pathology ; Chronic Disease ; Granuloma ; pathology ; Hearing Loss ; Hearing Loss, Conductive ; Humans ; Meningitis ; etiology ; Otitis Media ; complications ; pathology ; Retrospective Studies ; Tympanic Membrane