Objective To investigate the mechanism of cartilage injury and inflammation in the WHBE rabbit KOA model and the effect of platelet-rich fibrin releasates(PRFr)treatment on the KOA process,we established a WHBE rabbit KOA model by excision of medial collateral and partial patellar ligaments and administered a PRFr solution.Methods Twenty-four WHBE rabbits were randomly divided into three groups:normal control(NC)group(n=6),model(KOA)group(n=12),and cure(PRFr)group(n=6).KOA and PRFr groups were injected with 0.5 mL saline and PRFr into both joint cavities on 7 and 14 postoperative days,respectively.At 4 and 8 weeks of modeling,the knee joint grade scoring,X-ray imaging,and gross scoring were performed.Serum levels of IL-1β,TNF-α,and MMP-13 were measured by ELISA.At 4 weeks,6 animals in the KOA group were euthanized,and at 8 weeks,the remaining animals in each group were euthanized.Pathological sections were prepared after decalcification,and then HE,toluidine blue,and safranin O-fast green staining and immunohistochemical analysis of TGF-β,BMP3,and NF-κB were conducted.Results The Lequesne MG behavioral score,Mankin's score,and Pelletier score of WHBE rabbits after the operation were significantly increased compared with the NC group(P＜0.01).Pathological observations revealed surface defects of the cartilage and partial loss of chondrocytes.These result indicated that the KOA model was established successfully.In KOA rabbits,knee joint swelling,joint pain stimulation,and movement limitation were obvious.X-rays showed a high-density soft tissue shadow,indicating more joint effusion and a rough articular surface in general.After PRFr treatment,the serum levels of proinflammatory factors IL-1β,TNF-α,and MMP-13 in KOA model rabbits were significantly reversed(P＜0.05,P＜0.01).Additionally,the cartilage surface became smooth,and most chondrocytes were neatly distributed.Expression levels of TGF-β,BMP3,and NF-κB induced by KOA were also significantly decreased(P＜0.01).Conclusions We successfully established a KOA model in WHBE rabbits,and PRFr improved the cartilage injury and inflammation of the WHBE rabbit KOA model through TGF-β/BMP and NF-κB pathways.

Objective To establish a rat model of hyperuricemic nephropathy(HN)using a multifactorial induction method of potassium oxazinate combined with adenine and yeast feed to observe the intervention effect of Qiling granules(QLG).Methods Fifty-eight SPF-grade male SD rats were selected,and 10 rats were randomly allocated to the normal control(NC)group.The remaining rats were induced by multiple factors to establish HN rat models.After 2 weeks of modeling,submandibular blood samples were taken to detect serum UA,CREA,BUN,TG,and TC.Forty HN rats with bleeding clearance UA and body weight close to the mean were selected.They were randomly divided into a model(M)group,QLG low dose(QLG-L)groups,QLG high dose(QLG-H)group,and a positive control(PC)group,with 10 rats in each group,using a stratified randomization method.Each group was given corresponding drugs by gavage daily,and after continuous administration for 4 weeks,submandibular blood samples were taken to detect serum UA,CREA,BUN,TG,and TC.After euthanasia of the rats,liver tissue was taken to detect XOD and ADA activity.Renal tissue was taken for HE and Gomori hexamine silver staining,and the protein expression of GLUT9,OAT1,VCAM-1,and TGF-β in the kidneys was observed using immunohistochemistry and Western blot method.Results Compared with the NC group,the M group's serum levels of UA,CREA,BUN,TC,and TG,as well as liver XOD and ADA activities,were significantly increased(P＜0.01).The renal tissue of the model rats showed significant pathological changes.The area of renal tubules positive for urate and the expression of GLUT9,VCAM-1,and TGF-β proteins in the kidneys were significantly increased(P＜0.01,P＜0.05),while the expression of OAT1 was significantly reduced(P＜0.01).Compared with the M group,each treatment group showed significantly reduced serum UA levels,liver XOD,ADA activity,and renal VCAM-1 protein expression(P＜0.01,P＜0.05).The serum CREA and BUN levels and renal TGF-β protein expression of rats in the QLG-L group were significantly reduced(P＜0.05,P＜0.01).The serum CREA and BUN levels and renal GLUT9 protein expression of rats in the QLG-H group were also significantly reduced(P＜0.01,P＜0.05).The urate deposition and renal injury caused by each treatment were reduced to varying degrees,but there were no significant differences among groups(P＞0.05).Conclusions A stable HN rat model can be induced by gavage of potassium oxyzinate and adenine in combination with yeast feed.QLG can effectively treat HN by improving UA metabolic disorders,reducing the renal inflammation and urate deposition that cause renal damage in HN model rats.Its mechanism of action is related to a reduction in serum UA,CREA,BUN,and TG levels;liver XOD and ADA activities;and the expression of GLUT9,OAT1,VCAM-1,and TGF-β proteins in the kidneys.

Objective:To study the distribution and antibiotics resistance of the main pathogens of neonatal purulent meningitis in different regions of China.Methods:A retrospective descriptive clinical epidemiological study was conducted in children with neonatal purulent meningitis which admitted to 18 tertiary hospitals in different regions of China between January 2015 to December 2019. The test results of blood and cerebrospinal fluid, and drug sensitivity test results of the main pathogens were collected. The distributions of pathogenic bacteria in children with neonatal purulent meningitis in preterm and term infants, early and late onset infants, in Zhejiang Province and other regions outside Zhejiang Province, and in Wenzhou region and other regions of Zhejiang Province were analyzed. The chi-square test was used for statistical analysis.Results:A total of 210 neonatal purulent meningitis cases were collected. The common pathogens were Escherichia coli ( E. coli)(41.4%(87/210)) and Streptococcus agalactiae ( S. agalactiae)(27.1%(57/210)). The proportion of Gram-negative bacteria in preterm infants (77.6%(45/58)) with neonatal purulent meningitis was higher than that in term infants (47.4%(72/152)), and the difference was statistically significant ( χ2=15.54, P=0.001). There were no significant differences in the constituent ratios of E. coli (36.5%(31/85) vs 44.8%(56/125)) and S. agalactiae (24.7%(21/85) vs 28.8%(36/125)) between early onset and late onset cases (both P>0.05). The most common pathogen was E. coli in different regions, with 46.7%(64/137) in Zhejiang Province and 31.5%(23/73) in other regions outside Zhejiang Province. In Zhejiang Province, S. agalactiae was detected in 49 out of 137 cases (35.8%), which was significantly higher than other regions outside Zhejiang Province (11.0%(8/73)). The proportions of Klebsiella pneumoniae, and coagulase-negative Staphylococcus in other regions outside Zhejiang Province (17.8%(13/73) and 16.4%(12/73)) were both higher than those in Zhejiang Province (2.9%(4/137) and 5.1%(7/137)). The differences were all statistically significant ( χ2=14.82, 12.26 and 7.43, respectively, all P<0.05). The proportion of Gram-positive bacteria in Wenzhou City (60.8%(31/51)) was higher than that in other regions in Zhejiang Province (38.4%(33/86)), and the difference was statistically significant ( χ2=6.46, P=0.011). E. coli was sensitive to meropenem (0/45), and 74.4%(32/43) of them were resistant to ampicillin. E. coli had different degrees of resistance to other common cephalosporins, among which, cefotaxime had the highest resistance rate of 41.8%(23/55), followed by ceftriaxone (32.4%(23/71)). S. agalactiae was sensitive to penicillin, vancomycin and linezolid. Conclusions:The composition ratios of pathogenic bacteria of neonatal purulent meningitis are different in different regions of China. The most common pathogen is E. coli, which is sensitive to meropenem, while it has different degrees of resistance to other common cephalosporins, especially to cefotaxime.

Objective:To study the clinical characteristics of purulent meningitis complicated with hydrocephalus in neonates, and to analyze the risk factors of the disease.Methods:Neonates diagnosed with purulent meningitis complicated with hydrocephalus who hospitalized in the department of neonatology of the Second Affiliated Hospital of Wenzhou Medical University from January 2002 to August 2021 were selected as the case group. Neonates with positive pathogen cultures but no hydrocephalus during the same period were assigned by random number table method as the control group. The ratio of the control group and the case group was 2 ∶1. The clinical data such as bacteria distribution, cranial imaging, therapy and prognosis were compared between the two groups. The risk factors for hydrocephalus were predicted. Statistical analysis was conducted using chi-square test and multiple logistic regression analysis.Results:There were 33 cases in the case group and 66 cases in the control group. A total of 27 cases had confirmed pathogen results, of which 20 cases (74.1%) were Gram-negative bacteria and seven cases (25.9%) were Gram-positive bacteria. The time of diagnosis for hydrocephalus were 13.0(5.5, 28.5) days after the onset. Twenty-six cases received non-surgical treatment, while seven cases received surgery. The cure rate of case group was 42.4%(14/33), which was lower than that of control group (72.7%, 48/66), and the difference was statistically significant ( χ2=8.63, P=0.003). Univariate analysis showed that the incidences of protein>3 g/L in cerebrospinal fluid, glucose<2 mmol/L in cerebrospinal fluid, convulsions, central respiratory failure, intracranial hemorrhage and encephalomalacia in the case group were all higher than those in the control group, with statistical significance ( χ2=19.72, 12.04, 19.04, 5.73, 11.85 and 17.48, respectively, all P<0.050). Multivariate logistic regression analysis showed that convulsions (odds ratio ( OR)=4.476, 95% confidence interval ( CI) 1.091 to 18.363, P=0.037), intracranial hemorrhage ( OR=8.031, 95% CI 1.894 to 34.059, P=0.005) and encephalomalacia ( OR=35.189, 95% CI 2.954 to 419.150, P=0.005) were risk factors for neonatal purulent meningitis complicated with hydrocephalus. Conclusions:Gram-negative bacteria are common pathogen of neonatal purulent meningitis complicated with hydrocephalus. Convulsions, intracranial hemorrhage and encephalomalacia are important predictors for neonatal purulent meningitis complicated with hydrocephalus.

Objective:To summarize the clinical characteristics, diagnosis, treatment, and prognosis of neonatal meningitis caused by Mycoplasma hominis. Methods:We present the clinical data, diagnosis and treatment of a premature infant with Mycoplasma hominis meningitis who was admitted to the Department of Neonatology, the Second Affiliated Hospital of Wenzhou Medical University in June 2020. Relevant literature up to May 2021 was retrieved with the strategy of "( Mycoplasma hominis) AND (meningitis OR central nervous system OR cerebrospinal fluid) AND (newborn)" from CNKI, Wanfang, and PubMed database. The clinical manifestations, examinations, diagnosis, treatments and prognosis of cases with complete clinical data were summarized using two-sample rank sum test. Results:A premature female infant at gestational age of 27 +4 weeks presented with repeated low-grade fever and apnea since the 7 days of life. Cerebrospinal fluid testing in a local hospital showed neutrophil-based leukocytosis, which indicated purulent meningitis. However, empiric antibiotic treatment did not improve the infant's condition. The patient was transferred to our hospital due to dyspnea for 32 days and repeated fever for 25 days. Mycoplasma hominis was detected from the cerebrospinal fluid samples using metagenomic next generation sequencing (NGS). Treatment with erythromycin was ineffective, but the patient improved and discharged after changing to chloramphenicol for 18 d without any side effects. A total of 21 English articles were retrieved, and no Chinese literature was retrieved, involving 22 infants. Of the 23 cases including the present case, 14 were preterm, eight were term and one with no available data; 19 were born by vaginal delivery; the median age of onset was 11.0 d ( P25- P75: 7.0-18.0 d). The initial symptoms included fever, convulsions, irritability, and apnea. Blood routine examination showed elevated white blood cell count in ten cases and elevated C-reactive protein in seven cases. In the cerebrospinal fluid testing, white blood cell count increased in 19 cases, protein increased in 20 cases, and glucose decreased in 13 cases. Eight cases were confirmed by 16S RNA polymerase chain reaction amplification technology, seven by serum antibodies test, two cases by culture and microscopic findings, two cases by culture alone, one case by Mycoplasma kit, and one by NGS. The main treatment was the administration of tetracyclines, quinolones, chloramphenicol, lincosamides, etc. (alone or in combination). Two cases improved without using special anti- Mycoplasma drugs. Of the 23 patients, 15 had hydrocephalus, eight had intracranial hemorrhage, four had cerebral ischemic infarction, and two had cerebral abscess. Four cases had good prognosis,16 cases had adverse prognosis, and other three without available data. The median time to start sensitive antibiotic therapy in children with good prognosis was 4.5 d(3.6-5.0 d) after diagnosis, which was earlier than that in children with adverse prognosis [16.8 d (7.0-25.0 d)]( Z=－2.27, P=0.023). Conclusions:Mycoplasma hominis infection has non-specific clinical manifestations and should be considered for infants with intracranial infection that is not responding to empirical antibiotic treatment. NGS is helpful in detecting Mycoplasma hominis and chloramphenicol can be an option for the treatment.

Objective:To explore the effectiveness, safety and cost between urinary follicle stimulating hormone (uFSH) and recombinant follicle stimulating hormone (rFSH) in controlled ovarian stimulation (COS) in China.Methods:Data were collected from 16 reproductive centers in China covering oocytes collection time from May 1, 2015 to June 30, 2018. Eligible patients were over 18 years old, adopting COS with uFSH (uFSH group) or rFSH (rFSH group) as start gonadotropins (Gn), and using in vitro fertilization (IVF) and (or) intracytoplasmic sperm injection for fertilisation, excluding frozen embryo recovery cycle. Generalised estimating equation was used to address the violation of independency assumption between cycles due to multiple IVF cycles for one person and clustering nature of cycles carried out within one center. Controlling variables included age, body mass index, anti-Müllerian hormone level, cause of infertility, ovulation protocol, type of fertilisation, number of embryos transferred, number of days of Gn use.Results:Totally 102 061 cycles met eligibility criteria and were included in the analyses. In terms of effectiveness, after controlling relevant unbalanced baseline characteristics, compared with rFSH group, the high oocyte retrieval (>15 oocytes was considered high retrieval) rate of uFSH group significantly decreased in gonadotropin-releasing hormone agonist protocol ( OR=0.642, P<0.01) and in gonadotropin-releasing hormone antagonist protocol ( OR=0.556, P=0.001), but the clinical pregnancy rate per transfer cycle and the live birth rate per transfer cycle significantly increased ( OR=1.179, OR=1.169, both P<0.01) in both agonist and antagonist protocols. For safety, multiple analysis result demonstrated that in the agonist protocol, compared with rFSH group, the incidence of moderate to severe ovarian hyperstimulation syndrome of uFSH group significantly decreased ( OR=0.644, P=0.002). The differences in ectopic pregnancy rate and multiple pregnancy rate between the uFSH and rFSH groups were not significant ( P=0.890, P=0.470) in all patients. In terms of cost, compared with rFSH group, the uFSH group had lower total Gn costs for each patient ( P<0.01). Conclusion:For patients who underwent COS, uFSH has better safety, and economic profiles over rFSH in China.

Objective To study the clinical value of tumor necrosis factor-α (TNF-α) and resolvin D1 (RvD1) concentrations in cerebrospinal fluid (CSF) of neonatal purulent meningitis(NPM).Method From June 2016 to June 2017,neonates of suspected NPM admitted to the neonatology department of our hospital were studied prospectively.Their CSF was examined before the use of antibiotics.The patients were assigned into NPM group and non-NPM group.After 7 to 10 days of treatment,according to the clinical symptoms and the reexamination results of CSF,patients in the NPM group were further assigned into the improved group and the unimproved group.The levels of TNF-α and RvD1 in CSF were measured using enzyme-linked immunosorbent assay (ELISA) method,and SPSS 22.0 was used for statistical analysis.Result A total of 23 patients were included in the NPM group (18 in the improved group and 5 in the unimproved group) and 30 in the non-NPM group.The levels of TNF-α and RvD1 in the CSF of the NPM group were higher than the non-NPM group [TNF-α:(0.263 ±0.088) pg/ml vs.(0.087 ±0.001) pg/ml,RvD1:(2.017 ± 0.171) pg/ml vs.(0.563 ±0.048) pg/ml] (P ＜0.05).After 7 to 10 days of treatment,TNF-α and RvD1 decreased in the improved NPM group[TNF-α:0.083 (0.078,0.111) pg/ml vs.0.122 (0.098,0.214) pg/ml,RvD1:1.242 (0.740,2.098) pg/ml vs.1.791 (1.371,2.804) pg/ml] (P ＜ 0.05),and increased in the unimproved NPM group [TNF-α:2.239 (1.309,2.806) pg/ml vs.0.102 (0.100,1.312) pg/ml,RvD1:2.614 (1.265,2.940) pg/ml vs.0.139 (0.103,0.276) pg/ml] (P ＜ 0.05).The reexamination results of TNF-oα in the NPM group were lower than the examination results before the use of antibiotics of the non-NPM group,and RvD1 higher than the non-NPM group (P ＜ 0.05).Conclusion TNF-α and RvD1 in CSF have clinical value for the early diagnosis and therapeutic evaluation of NPM.

Objective To detect the expression and clinic significance of long non-coding RNA ZEB1-AS1 in breast cancer. Methods A total of 130 patients with breast cancer in Baoji Central Hospital of Shaanxi Province from June 2007 to April 2015 were selected. Quantitative real-time PCR(qRT-PCR)was used to detect the expression level of ZEB1-AS1 in breast cancer tissues and corresponding normal tissues,and the rela-tionships between the expression level of ZEB1-AS1 and the clinic characteristics of the patients and their overall survival time were analyzed. siRNA was used to disturb the expression of ZEB1-AS1. CCK-8 assay, clone formation assay and Transwell assay were used to detect the proliferation,cloning ability and migration of breast cancer MCF-7 cells in control group,siRNA-1 group and siRNA-2 group. Results The expression level of ZEB1-AS1 in breast cancer tissues was higher than that in corresponding normal tissues[M(QR ):0. 0016 (0. 0051)vs. 0. 0009(0. 0015);Z = - 4. 426,P < 0. 001]. The higher expression of ZEB1-AS1 was correlated with lymphatic metastasis(χ2 = 9. 148,P = 0. 027),negative human epidermal growth factor recep-tor 2(χ2 = 5. 039,P = 0. 025),triple negative breast cancer(χ2 = 4. 597,P = 0. 032). The patients with the higher expression of ZEB1-AS1 had a shorter overall survival time compared with the patients with the lower expression of ZEB1-AS1(χ2 = 14. 340,P < 0. 001). CCK-8 assay showed that knock down of ZEB1-AS1 after 72 h,the absorbance values of the control group,siRNA-1 group and siRNA-2 group were 0. 605 ± 0. 049, 0. 488 ± 0. 054,0. 417 ± 0. 038 respectively,with a statistically significant different( F = 15. 936,P <0. 001),and the two siRNA groups were significantly inhibited in cell proliferation compared with the control group(both P < 0. 05). The colonies of the control group,siRNA-1 group and siRNA-2 group were 297. 5 ± 11. 4,192. 0 ± 12. 1,204. 8 ± 12. 8 respectively,with a statistically significant different(F = 112. 526,P <0. 001),and the two siRNA groups were significantly inhibited in the cell clone compared with the control group(both P < 0. 001). The migratory cells numbers of the control group,siRNA-1 group and siRNA-2 group were 184. 5 ± 8. 6,147. 5 ± 18. 6,57. 6 ± 7. 3 respectively,with a statistically significant different( F =12. 409,P = 0. 001),and the two siRNA groups were significantly inhibited in the cell migration(both P <0. 001). Conclusion ZEB1-AS1 is overexpressed in breast cancer,overexpression of ZEB1-AS1 induces a shorter overall survival in breast cancer patients,and knock down of ZEB1-AS1 can inhibit the proliferation, migration and colony formation ability of the breast cancer cell line.

Objective: To investigate the predictive factors of mortality in extremely preterm infants. Methods: The retrospective case-control study was accomplished in the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University. A total of 268 extremely preterm infants seen from January 1, 1999 to December 31, 2015 were divided into survival group (192 cases) and death group (76 cases). The potential predictive factors of mortality were identified by univariate analysis, and then analyzed by multivariate unconditional Logistic regression analysis. The mortality and predictive factors were also compared between two time periods, which were January 1, 1999 to December 31, 2007 (65 cases) and January 1, 2008 to December 31, 2015 (203 cases). Results: The median gestational age (GA) of extremely preterm infants was 27 weeks (23⁺³-27⁺⁶ weeks). The mortality was higher in infants with GA of 25-<26 weeks (OR=2.659, 95% CI: 1.211-5.840) and<25 weeks (OR=10.029, 95% CI: 3.266-30.792) compared to that in infants with GA> 26 weeks. From January 1, 2008 to December 31, 2015, the number of extremely preterm infants was increased significantly compared to the previous 9 years, while the mortality decreased significantly (OR=0.490, 95% CI: 0.272-0.884). Multivariate unconditional Logistic regression analysis showed that GA below 25 weeks (OR=6.033, 95% CI: 1.393-26.133), lower birth weight (OR=0.997, 95% CI: 0.995-1.000), stage Ⅲ necrotizing enterocolitis (NEC) (OR=15.907, 95% CI: 3.613-70.033), grade Ⅰ and Ⅱ intraventricular hemorrhage (IVH) (OR=0.260, 95% CI: 0.117-0.575) and dependence on invasive mechanical ventilation (OR=3.630, 95% CI: 1.111-11.867) were predictive factors of mortality in extremely preterm infants. Conclusions GA below 25 weeks, lower birth weight, stage Ⅲ NEC and dependence on invasive mechanical ventilation are risk factors of mortality in extremely preterm infants. But grade ⅠandⅡ IVH is protective factor.

Objectives To investigate the risk factors for rheumatoid arthritis in middle-aged and elderly residents in Luohe City,Henan Province.Methods Survey questionnaires and measurements of biochemical parameters were conducted in individuals 45 years old or above,using the two-stage cluster sampling method.Risk factors for rheumatoid arthritis were evaluated according to the criteria for the classification of RA by the American College of Rheumatology and European League Against Rheumatism(ACR/EULAR) (2010).Results Among the 8 610 residents covered by the survey,8,274 people responded,representing a response rate of 96.1％.There were 3 361 male (40.62％)and 4 913 female(59.38％)respondents,with an average age of 61.4±7.4(45-91)years.The age group of 60-64 years had the highest rates of joint involvement,with those scoring 2,3 and 5 points at 2.90％,2.02％ and 0.26％,respectively.The rheumatoid factor (RF)and the anti-cycliccitrullinatedpeptide(anti-CCP)antibody titer showed skewed distributions.The low titer-positive rate of RF and anti-CCP antibody,the ESR and CRP in female residents were evidently higher than in male residents(each P＜0.05).Smoking was an independent factor for RA(OR:1.79,95 ％CI:1.34～ 3.41,P＜0.01).The risk for RA occurrence increased with the frequency of drinking ＞1 time/d(OR:6.71,95 ％ CI:0.88 ～ 51.23,P＜ 0.01).The prevalence of RA was significantly higher in rural areas than in urban and suburb areas(0.93％ to 0.48％ and 0.53％,each P＜0.05).Living on higher floors was a protective factor for RA (OR:0.61,95 ％ CI:0.36 ～ 0.94,P =0.036).Family history was an independent risk factor for RA (OR:3.09,95 ％ CI:1.53 ～ 6.27,P ＜ 0.01),and being first degree relatives of RA patients was a risk factor(OR=6.45,95 ％ CI:1.67～ 17.83,P＜0.01).Multiple factor analysis showed that female gender,first floor of residential buildings,smoking and genetics were key risk factors for RA.Conclusions The risk factors for rheumatoid arthritis in middle-aged and elderly residents in Luohe City of Henan Province are female gender,smoking and genetics.