Objective To establish a genetic monitoring method for laboratory quails.Methods Quail microsatellite loci were searched in the literature,and microsatellite DNA loci suitable for quails were screened by an interspecific transfer method in closely related species,namely chickens and ducks.Quail liver DNA was extracted as a template,and the corresponding loci were screened by PCR amplification and agarose gel electrophoresis.On the basis of amplification of the selected microsatellite loci,the number of alleles,polymorphisms,and microsatellite loci combinations for quail genetic quality detection were selected and detection method were developed.Results We preliminary determined 23 microsatellite loci for genetic monitoring of closed-colony laboratory quails.Conclusions A genetic monitoring method for laboratory quails was preliminary developed.

Objective:To observe the recurrence condition of hepatitis B in different risk groups after liver transplantation in an attempt to provide useful information on whether to discontinue hepatitis B immunoglobulin (HBIG) in the future at an early stage.Methods:The patient population was divided into high, low-risk, and special groups [especially primary hepatocellular carcinoma (HCC)] according to the guidelines for the prevention and treatment of hepatitis B recurrence after liver transplantation. The recurrence condition and risk factors in this population were observed for hepatitis B. Measurement data were analyzed using a t-test and a rank-sum test. Count data were compared using a χ2 test between groups. Results:This study finally included 532 hepatitis B-related liver transplant cases. A total of 35 cases had HBV recurrence after liver transplantation, including 34 cases that were HBsAg positive, one case that was HBsAg negative, and 10 cases that were hepatitis B virus (HBV) DNA positive. The overall HBV recurrence rate was 6.6%. The recurrence rate of HBV was 9.2% and 4.8% in the high- and low-risk HBV DNA positive and negative groups before surgery ( P = 0.057). Among the 293 cases diagnosed with HCC before liver transplantation, 30 had hepatitis B recurrence after surgery, with a recurrence rate of 10.2%. The independent related factors for the recurrence of hepatitis B in patients with HCC after liver transplantation were HCC recurrence ( HR =181.92, 95% CI 15.99~2 069.96, P < 0.001), a high postoperative dose of mycophenolate mofetil dispersible tablets (MMF) ( HR =5.190, 95% CI 1.289~20.889, P = 0.020), and a high dosage of HBIG ( HR = 1.012, 95% CI 1.001~1.023, P = 0.035). Among the 239 cases who were non-HCC before liver transplantation, five cases (recurrence rate of 2.1%) arouse postoperative hepatitis B recurrence. Lamivudine was used in all cases, combined with on-demand HBIG prophylaxis after surgery. There was no hepatitis B recurrence in non-HCC patients who treated with entecavir combined with HBIG after surgery. Conclusion:High-barrier-to-resistance nucleotide analogues combined with long-term HBIG have a good effect on preventing the recurrence of hepatitis B after liver transplantation. The discontinuation of HBIG may be considered at an early stage after administration of a high-barrier-to-resistance nucleotide analogue in low-risk patients. Domestically, the HBV infection rate is high, so further research is still required to explore the timing of HBIG discontinuation for high-risk patients, especially those with HCC.

Objective To investigate the clinical characteristics and risk factors of mixed connective tissue disease associated with pulmonary arterial hypertension(MCTD-PAH).Methods Twelve MCTD-PAH patients diagnosed by right heart catheterization(RHC)at Tianjin Medical University General Hospital were retrospectively included,and 36 MCTD patients without pulmonary arterial hypertension(MCTD-non-PAH)were randomly selected from the same period of hospitalization based on gender and age.The clinical features and auxiliary examination of the two groups were compared,and the survival status of the two groups was compared.Results The proportion of dyspnea after activity,myositis and pericardial effusion were higher in the MCTD-PAH group than those of the control group.Serum sedimentation rate and immunoglobulin G(IgG)levels were higher in the MCTD-PAH group.Multivariate Logistic regression analysis showed that dyspnea after activity and high level of IgG were risk factors for predicting the occurrence of PAH in MCTD.Three patients(16.7%)died in the MCTD-PAH group,and no patients died in the control group.Conclusion Pulmonary arterial hypertension is one of the serious complications of MCTD.MCTD patients have shortness of breath after activity and high level of IgG should be wary of concomitant PAH.

Objective:To investigate the effects of Longmu Anshen prescription on microglia(MG)polarization in the medial prefrontal cortex(mPFC)and anxiety behavior of chronic restraint stress(CRS)mice.Methods:Forty-eight healthy male KM mice were randomly divided into normal group,paroxetine hydrochloride group and Longmu Anshen prescription high,middle and low dose groups.From the 15th day,after 30 minutes of CRS every day,the normal group and the model were given distilled water by gavage,the paroxetine hydrochloride group was given paroxetine hydrochloride(2.6 mg/kg)by gavage,and the traditional Chinese medicine group was given Longmu Anshen prescription at high,medium and low doses(4.16,2.08,1.04 g/kg)by gavage for 21 days.The changes of anxiety behavior in mice were evaluated by light-dark boxes experiment and elevated plus maze;ELISA was used to detect IL-1β,IL-6,TNF-α and IL-10 levels;protein expression of inducible nitric oxide synthase(iNOS)and arginase-1(Arg-1)in mice mPFC tissue were detected by Western blot;mRNA expression of ionized calcium binding adaptor molecule-1(Iba-1),iNOS and Arg-1 in mice mPFC tissue were detected by RT-PCR;the protein expression of Iba-1 in mice hippocampal tissue was detected by immuno-histochemistry(IHC).Results:Compared with normal group,the residence time in the light boxes,the number of times of shuttle between the light-dark boxes,the percentage of times of entering the open arm(OE%)and the percentage of time spent in the open arm(OT%)of mice in model group were significantly reduced(P<0.01);moreover,in the mPFC tissue of the model group mice,the protein and mRNA expressions of Iba-1 and iNOS were significantly increased(P<0.01),and the levels of IL-1β,IL-6 and TNF-α were significantly increased(P<0.01),while the protein and mRNA expressions of Arg-1 were significantly decreased(P<0.01),and the level of IL-10 was significantly decreased(P<0.01).Compared with model group,the residence time in the light boxes,the number of shuttles between the light-dark boxes,the OE%and OT%of mice in the paroxetine hydrochloride group and the high and medium dose groups of Longmu Anshen prescription were significantly increased(P<0.01);the expression levels of Iba-1 and iNOS protein and mRNA in mPFC tissue were significantly decreased(P<0.05,P<0.01),and the levels of IL-1β,IL-6 and TNF-α were significantly decreased(P<0.01),while the expression levels of Arg-1 protein and mRNA were significantly increased(P<0.01).IL-10 level was significantly increased(P<0.01).Conclusion:Longmu Anshen prescription can inhibit the activation of MG in mPFC tissues of CRS mice and may reduce nerve inflammation and increase the level of anti-inflammatory cytokines by regulating the polarization phenotype of M1/M2,thereby ameliorate CRS-induced anxiety behavior.

Objective To investigate the infection status and drug resistance of urogenital tract mycoplasma in Lianyungang district,Jiangsu.Methods Based on retrospective collection of 22 889 cases of suspected mycoplasma genital infection in Lianyungang Mater-nity and Child Health Hospital from 2015 to 2022,the infection and drug resistance rate of genital mycoplasma were analyzed.Results A total of 8 943 genital mycoplasma positive specimens were detected.Among them,the infection rate of Ureaplasma urealyticum(Uu)was the highest,accounting for 33.64％(7 700/22 889),which was significantly higher than that of Mycoplasma hominis(Mh),3.91％(894/22 889),and that of mixed(Uu+Mh)type,1.52％(349/22 889).The difference was statistically significant(P＜0.05).The infection rates of genital mycoplasma and Uu showed increasing trend from 2015 to 2022.The overall infection rates of genital mycoplasma in females were 42.42％,which was significantly higher than that in males of 22.47％.The highest infection rate of 40.84％(6 108/14 955)was found in the group of 20 to 40 years old as compared to those younger than 20 years old as well as those over 40 years old.The genital mycoplasma was highly sensitive to doxycycline,minocycline and josamycin,and the drug resistance rate was lower than 5.00％.From 2015 to 2022,there was an increasing trend of resistance rates to erythromycin,ofloxacin and ciprofloxa-cin in Uu-infected patients(P＜0.05)and erythromycin,roxithromycin,azithromycin and ofloxacin in Mh-infected patients(P＜0.05).Conclusion The incidence of urogenital tract mycoplasma infection in Lianyungang district shows a gradual increase for recent years.Doxycycline,minocycline and josamycin are suitable as the first choice for the treatment of genital mycoplasma infection.Most quinolo-ne drugs have shown high resistance rate,indicating that this type of drugs are possibly overused.The strengthened relevant monitoring of antibacterial drugs should be necessary.

Stroke is a prevalent acute cerebrovascular disease associated with significant morbidity and mortality,which highly demand effective prevention and treatment strategies.Glucagon-like peptide-1 receptor agonists(GLP-1RA)is a novel type of antidiabetic drug that exerts hypoglycemic and weight loss effects on GLP-1 receptors.Additionally,GLP-1RA possess the potential to reduce infarction size and promote nerve recovery by inhibiting inflammation,oxidative stress,apoptosis,and improving blood-brain barrier permeability and other pathologies.This paper provides a comprehensive summary of the role of GLP-1 in stroke occurrence while also explores the underlying mechanisms by which GLP-1RA influences stroke pathogenesis.Furthermore,it briefly reviews the therapeutic efficacy of GLP-RA in managing stroke.

Objective:To explore the clinical characteristics, chest imaging manifestations, RAPID score and therapeutic situation in patients with parapneumonic pleural effusion (PPE) caused by streptococcus anginosus group (SAG), in order to provide help for the early diagnosis and treatment in clinical practices. Methods:The clinical data of 39 patients with PPE caused by SAG from January 2015 to May 2020 in Affiliated Hospital of Jining Medical University and Jining First People′s Hospital were retrospectively analyzed. The patients were classified by RAPID score.Results:Among 39 cases, males was in 31 cases (79.5%), females in 8 cases (20.5%), and aged 46 to 89 (65.31±10.53) years old. Fever was in 27 cases (69.2%), chest pain in 19 cases (48.7%), and dyspnea in 18 cases (46.2%). The chest CT findings showed consolidation shadows was in 30 cases (76.9%), encapsulated pleural effusion in 21 cases (53.8%), ground glass shadow in 18 cases (46.2%), nodules in 12 cases (30.8%), atelectasis in 8 cases (20.5%), and pneumothorax in 5 cases (12.8%). The complexity PPE was in 23 cases (59.0%), and empyema in 16 cases (41.0%). The microbiological culture results showed that streptococcus constellatus was detected in 25 cases (64.1%), streptococcus anginosus in 13 cases (33.3%), and streptococcus intermadius in 1 case (2.6%). After comprehensive treatment, 36 cases (92.3%) were improved, 3 cases (7.7%) died. According to the RAPID score, low-risk was in 13 cases (33.3%), intermediate-risk in 16 cases (41.0%), and high-risk in 10 cases (25.7%). The RAPID score in patients with low-risk, intermediate-risk and high-risk was (1.85 ± 0.38), (3.43 ± 0.51) and (5.30 ± 0.67) scores, and there was statistical difference ( F = 124.88, P<0.05). the length of stay in patients with low-risk, intermediate-risk and high-risk of RAPID score was (16.84 ± 5.57), (16.56 ± 7.05) and (28.20 ± 17.97) d, and there was statistical difference ( F = 4.41, P<0.05); the length of stay in patients with high-risk was significantly longer than that in patients with low-risk and intermediate-risk, and there was statistical difference ( P<0.05), there was no statistical difference between intermediate-risk patients and low-risk patients ( P>0.05). Conclusions:SAG, as important pathogens for the PPE, tends to induce CPPE and even pyopneumothorax. Clinical manifestations and imaging are not specific, which should be payed attention in clinical work. The patients with high-risk of RAPID score have more serious condition and worse prognosis.

Objective:To explore the safety of inactivating coronavirus disease 2019(covid-19)vaccine in liver transplantation(LT)recipients.Methods:Retrospective analysis was performed for clinical data of 151 LT recipients from March 2003 to October 2019.They had stable conditions and completed the course of covid-19 vaccine.Frequencies of pain at injection site, fatigue, headache and pruritus after vaccination were recorded.The safety profiles were compared between recipients with and without local and general adverse reactions after vaccination.At the same time, recipients completing two doses of covid-19 vaccines were grouped.According to vaccine companies, they were classified into Sinovac Biotech Ltd and Beijing Biological.Based upon more than or less than 60 years, they were grouped into <60 years and ≥60 years.The safety profiles of inactivating COVID-19 vaccine were compared in subgroups.Results:Among 151 eligible LT recipients, 98 of them were in group of age <60 years and 53 in group of age >60 years.The median period between vaccination and LT was 8.44(4.37, 12.39)years and the median concentration of tacrolimus 2.5(1.8, 3.9)ng/L.Eighty-three cases completed two doses of Sinovac Biotech Ltd(Sinovac Biotech Ltd group)and 40 cases Beijing Biological(Beijing Biological group); 14 cases had combined course of Sinovac Biotech Ltd and Beijing Biological, four recipients were vaccinated with inactivated vaccine from other companies and ten recipients did not know their inactivated vaccine' companies.After immunization, 24/151(15.9%)recipients had a local and general adverse reaction.The prevalence of pain at injection site, fatigue, headache and pruritus was 9.9%( n=15), 5.2%( n=8), 1.3%( n=2)and 0.7%( n=1)respectively.No significant differences existed in age( P=0.602), gender( P=0.752), period after LT( P=0.890), trough concentration of tacrolimus( P=0.377)or versions of covid-19 vaccine( P=0.582)between 24 cases with general adverse reaction and 127 without.Local and general reactions occurred in 16/83(19.3%)in Sinovac group and 5/40(12.5%)in Beijing Biological.There was no significant inter-group difference( P=0.769). There were 98 cases(64.9%)in <60 years group, 17 cases(17.3%)had local and general reaction, 53 cases(35.1%)in ≥60 years group and 7 cases(13.2%)had a local and systemic reaction.There was no significant inter-group difference( P=0.507). Conclusions:Covid-19 vaccine is safe for long-term survival LT recipients with normal liver function.Few participants present with mild fatigue and pain at injection site.

Objective:To explore the role and mechanism of exogenous active peptide spexin in regulating insulin resistance in adipose tissue.Methods:High-fat diet induced obesity model(DIO) mice and diabetic model(db/db) mice were given intraperitoneal injection of spexin(50 μg/kg) for 3 consecutive weeks, while each control group was given an equal volume of saline. After the intervention, the body weight, visceral fat weight and plasma biochemical indexes of the mice in each group were analyzed. Real-time fluorescence quantitative PCR was used to detect mRNA levels of Krüppel-like transcription factor 9(KLF9), peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α) and glucose transporter 4(GLUT4); Western blotting was used to detect the protein levels of KLF9, PGC-1α, GLUT4 and p-p38/p38 in adipose tissue.Results:Compared with DIO control mice, spexin-treated DIO mice showed a significant reduction in body weight ( P=0.043), adiposity ( P<0.001), glucose ( P<0.001), insulin ( P=0.008), HOMA-IR ( P<0.001) and elevation in glucose tolerance ( P=0.006) and insulin tolerance levels ( P=0.002). Compared with db/db control mice, spexin-treated db/db mice showed a significant reduction in body weight ( P<0.001), adiposity ( P<0.001), glucose ( P=0.041), insulin ( P=0.009), HOMA-IR ( P=0.007) and elevation in glucose tolerance ( P=0.008) and insulin tolerance levels ( P=0.031). In addition, the gene and protein levels of KLF9, PGC-1α and GLUT4 were significantly increased in the adipose tissue of spexin-treated DIO mice compared with DIO control mice [genes ( P<0.001, P<0.001, P=0.005); proteins ( P=0.047, P=0.022, P=0.001)], while p-p38/p38 protein levels were significantly decreased in the adipose tissue of spexin-treated DIO mice compared with DIO control mice ( P=0.002). Moreover, the gene and protein levels of KLF9, PGC-1α and GLUT4 were significantly increased in the adipose tissue of spexin-treated db/db mice compared with db/db control mice [genes ( P<0.001, P<0.001, and P=0.005); proteins ( P=0.001, P=0.004, and P<0.001)], while p-p38/p38 protein levels were significantly decreased in the adipose tissue of spexin-treated db/db mice compared with db/db control mice ( P=0.001). Conclusion:These results suggested that spexin may play a role in ameliorating adipose tissue insulin resistance in DIO mice and db/db mice through regulating p38MAPK-mediated inflammation and KLF9-PGC-1α-GLUT4 pathway-mediated glucose uptake.

OBJECTIVE To establish HPLC characteristic chro matogram of Jianpi yifei biyan prescription standard decoction ， to select the quality control index components and determine their contents. METHODS HPLC method combined with Similarity Evaluation System of TCM Chromatographic Fingerprint （2004 edition）were used to establish the characteristic chromatogram of 10 batches of Jianpi yifei biyan prescription standard decoction ；the similarity evaluation and common peaks identification were also carried out. Using common peak area of characteristic chromatogram as variables ，SPSS 26.0 software and SIMCA 14.1 software were used to perfor m cluster analysis （CA），principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）；differential components with variable important i n pro jection（VIP）value greater than 1.5 were screened；the contents of cimifugin and differential components were determined by the same method. RESULTS A total of 24 common characteristic peaks were identified ， and the similarities of 10 batches of samples were higher than 0.960；eight characteristic peaks were identified by comparison with reference substance. CA and PCA results revealed that the samples were classified into 3 categories.OPLS-DA analysis showed that 3 components with VIP value greater than 1.5， which were prim-O-glucosylcimifugin （peak 2），calycosin 7-O-β-D-glucopyranoside （peak 4） and 5-O-methylvisammioside （peak 6） in descending order. The linear ranges of prim- O- glucosylcimifugin，calycosin 7-O-β-D-glucopyranoside，cimifugin and 5-O-methylvisammioside were 0.010 7-0.213 0，0.007 8- 0.156 0，0.008 0-0.160 0，0.009 8-0.195 0 μg（r＞0.999），respectively. RSD values of precision ，repeatability and stability tests （24 h） were all less than 2％. Average recoveries were 105.98％（RSD＝1.75％，n＝6），98.06％（RSD＝3.87％，n＝6），96.38％（RSD＝ 4.03％ ，n＝6） and 104.17％（RSD＝1.27％ ，n＝6）. The contents of the above 4 components in 10 batches of samples were 12.12-18.87，3.86-6.40，3.10-4.27 and 11.17-15.79 μ g/mL，respectively. CONCLUSIONS The established HPLC characteristic chromatographic method is stable and feasible ，it can be used for the quality control of Jianpi yifei biyan prescription standard decoction. Prim- O-glucosylcimifugin，calycosin 7-O-β-D-glucopyranoside，cimifugin and 5-O-methylvisammioside can be used as the index components for quality control of the standard decoction.