[ ABSTRACT] AIM: To investigate the differential expression of human leukocyte antigen-G ( HLA-G) isoforms and its receptors in human monocyte line THP-1 after human cytomegalovirus ( HCMV) infection for exploring the role of HLA-G in HCMV escaping the immune response of the organism .METHODS: THP-1 cells were infected with HCMV Towne strain.The expression of HLA-G isoforms at mRNA and protein levels was determined by RT-PCR and Western blot, respectively.The surface expression of HLA-G and its receptors ILT2/ILT4 and the cell viability were analyzed by flow cytometry.The levels of soluble HLA-G (sHLA-G) and IL-10 were measured by ELISA.RESULTS:After infection of the THP-1 cells with HCMV , no obvious apoptosis in the cells was observed , and the viability of the cells was high .A significant up-regulation of HLA-G1,-G3,-G4 and-G5 at mRNA expression level 1 d after infection was found , while the protein expression of HLA-G1 and HLA-G5 isoforms was mainly detected .The expression of HLA-G/ILT2/ILT4 was evi-dently up-regulated 1 d after infection .The level of sHLA-G was significantly increased 1 d after infection as compared with control group (P<0.01).The expression of IL-10 was obviously up-regulated 1 d post-infection as compared with control group.CONCLUSION:The differential expression of HLA-G isoforms and secretion of the receptors ILT 2/ILT4 and IL-10 in the THP-1 cells are induced after HCMV infection .This study provides experimental evidence for evaluating the immune mechanism of HCMV infection .

Objective To investigate the development of early myocardial perfusion with myocardial contrast echocardiography (MCE) combined with dipyridamole stress echocardiography in diabetic rats . Methods The diabetes mellitus (DM) group comprised 40 male diabetic rats ,induced with streptozotocin . The control group comprised 40 normal male rats ,comparable body weights with the DM group .The DM group was divided into four subgroups (0 week ,2 weeks ,4 weeks and 8 weeks after diabetic model established) and the control group was also divided into four subgroups matched with the DM group .Each rat was performed with conventional echocardiography ,MCE at baseline and after dipyridamole stress .The reserve parameters were compared between the control group and the DM group .In addition ,the differences among four subgroups in the control group and the DM group were compared ,respectively .Results MCE demonstrated that the 4 weeks and 8 weeks DM subgroup had lower myocardial blood velocity reserve and myocardial blood flow reserve than the control subgroup .The myocardial blood volume reserve was reduced in the 8 weeks DM subgroup ,too .Conclusions The impairment of myocardial perfusion in the DM rats are detected earlier with the MCE combined with dipyridamole stress .

To evaluate the value of serum human epididymis protein 4(HE4),cancer embryonic antigen(CEA), carbohydrate antigen(CA125,CA19-9,CA153),alpha fetoprotein(AFP) and β-human chorionic gonadotrophin (β-HCG) in ovarian cancer early diagnosis.Methods:Abbott Architect i2000SR was used to detect the levels of preoperative serum tumor markers (n=7) in ovarian cancer group,benign ovarian tumor group and the normal control (each n=60).Results:The levels of serum HE4,CA125, CA199,CEA,CA153,AFP and β-HCG in ovarian cancer group were markedly higher than the benign ovarian tumor group and the control group.There was statistically significant difference ( P<0.05 ).The diagnosis of ovarian cancer tumor markers were high specificity,but sensitivity was not high except HE4 (76.67%) and CA125 (68.33%).With Youden Index,the area under curve of ROC,the HE4 was demonstrated higher specificity ,sensitivity and accuracy.There was also medium diagnostic value of CA 125 ,CA199 and CEA for ovarian cancer.The sensitivity of ovarian cancer diagnosis is raised by combined assay with serum tumor marker ,but the specificity would be reduced accordingly.HE4, CA125, CA199 and CEA combined detection is considered the best.Conclusion:CA153,AFP and β-HCG have lowered diagnosis for ovarian cancer.HE4, CA125, CA199 and CEA combined detection could significantly improve specificity ,sensitivity and accuracy in the early diagnosis of ovarian cancer.

Objective:To analyze clinical characteristics of cutaneous delayed-type hypersensitivity caused by injection of equine tetanus antitoxin (TAT) or equine anti-tetanus immunoglobulin F (ab′) 2. Methods:Clinical data were collected from 181 outpatients or inpatients with cutaneous delayed-type hypersensitivity caused by injection of equine TAT or anti-tetanus immunoglobulin from 2008 to 2020, and retrospectively analyzed.Results:Before the injection of equine TAT or anti-tetanus immunoglobulin, skin test was negative in 171 (94.47%) of the 181 patients, and the 10 (5.53%) patients with positive skin test responses received desensitization injection. Among the 181 patients, there were 118 males and 63 females aged from 11 to 68 years, with the disease duration of 1 to 7 days and alatency period of 4 to 14 days. There was no significant difference in the clinical manifestations between the patients receiving injection of TAT (130 cases) and those receiving injection of equine anti-tetanus immunoglobulin (51 cases) . Urticaria-like rashes were the main clinical manifestation, and infiltrative erythema occurred at the injection site in 12 patients, of whom 10 developed generalized urticaria all over the body. Of the 181 patients, 163 (90.06%) presented with generalized skin rashes, and 56 (30.94%) had systemic symptoms such as chest tightness, fever, etc, of whom 15 (26.79%) had a history of allergies and 6 with severe symptoms had no history of allergies. Thirty-four (18.78%) patients had single or multiple laboratory abnormalities, such as increased white blood cell counts, elevated C-reactive protein level and urinary glucose, and presence of occult blood in urine. All cases responded well to the treatment with antihistamines and glucocorticoids. The treatment duration ranged from 3 to 10 days, and the outcome was good.Conclusion:TAT-or anti-tetanus immunoglobulin-induced cutaneous delayed-type hypersensitivity may still occur in patients with a negative skin test or after desensitization treatment, and mainly manifests as urticaria-like rashes.

Objective To assess the myocardial function with early insulin treatment by velocity vector imaging (VVI) combined with dipyridamole stress in diabetic rats.Methods The 30 rats comprised 20 diabetic rats and 10 normal rats in the study.The 20 diabetic rats were divided into the diabetes mellitus(DM) group and the treatment group.Every rat in the treatment group was hypodermic injected with protamime biosynthetic human insulin after the diabetic model established.The control group was including 10 normal rats of the same age.After 8 weeks,each rat was performed with VVI at baseline and after dipyridamole stress.Results At baseline VVI,the parameters in the DM and the treatment groups were reduced compared with those in the control group.The systolic velocity,circumferential strain,systolic and diastolic circumferential strain rate in the treatment group were increased significantly compared with those in the DM group.After stress,the VVI reserve parameters in the treatment group were higher than those in the DM group.Conclusions The myocardial function are improved with early insulin treatment in the diabetic rats.

Objective To study effects ofXiangsha Liujunzi Decoction (XSLJZ) on gastric emptying function, pepsin activity and expression of HIF-1α gene and protein of gastric mucosa in chronic atrophic gastritis (CAG) of rats with deficiency of spleen and stomach.Methods Rats were randomly divided into normal group and model group. The models of CAG rats with deficiency of spleen and stomach type were induced by synthetic methods. After the modeling, models rats were divided into model group, positive control group and XSLJZ high-, medium-, and low-dose groups. Normal group and model group were given 10 mL/kg distilled water every day for gavage; XSLJZ high-, medium-, and low-dose groups were given 24, 12, and 6 g/kg XSLJZ Decoction for gavage; positive control group was given 0.30 g/kg Vatacoenayme for gavage, for successive 120 d. Gastric emptying function and pepsin activity were detected, and HIF-1α gene and protein expression in gastric mucosa were detected by RT-qPCR and IHC. Results Compared with the normal group, the gastric emptying function and pepsin activity in the model group were much lower (P<0.01); expressions of HIF-1α gene and protein in gastric tissues was much higher (P<0.01). Compared with model group, XSLJZ could increase the gastric emptying function and pepsin activity significantly (P<0.05), and decrease expressions of HIF-1α gene and protein (P<0.05).Conclusion XSLJZ has functions of improving the gastric emptying function, promoting pepsin activity, and reducing expressions of HIF-1α gene and protein.

Objective: To learn the toxicity of Dendrobium officinale flowers to pregnant rats ( P, F1 ) and offspring rats ( F1, F2 ) before birth, so as to provide toxicological evidence for the safety assessment. Methods : The rats were divided into four groups with 20 female rats and 10 male each. The rats in three dose groups were fed with Dendrobium officinale flowers at the dose of 2.0, 4.0, 6.4g/kgbw. After two generation, the F1a and F2a rats were fed with basal diet; F1b and F2b rats were fed with Dendrobium officinale flowers. The body weights and total weight gains during the gestation, the conception rates, the pregnancy rates, the birth weights and survival rates of offspring rats were examined. Results: There were no statistically significant differences in the body weights and total weight gains during the gestation, the conception rates, and the pregnancy rates in pregnant rats ( P, F1 ) among the four groups ( P>0.05 ). There were also no statistically significant differences in the survival rates and live birth rates in offspring rats (F1, F2) between the dose groups and the control group ( P>0.05 ). Conclusions Dendrobium officinale flowers did not show obviously adverse effects on pregnant rats ( P, F1 ) and offspring rats ( F1, F2 ) before birth.

Objective: To learn the effects of Dendrobium officinale flowers on testivular tissue morphology and sperm quality in parent and offspring rats,so as to provide reference for safety evaluation of Dendrobium officinale flowers. Methods: The 40 SD rats was randomly divided into the low-,middle-,high-dose and the control group,given 2.0,4.0,6.4 and 0 g/kgbw Dendrobium officinale flowers,respectively. After three months,the body weight,mass and organ/body coefficients of testis and epididymis of parent （P） and offspring （F1,F2） rats were measured;the number,activity and deformity of sperms were observed under microscope;the changes of testis and epididymis were observed by hematoxylin eosin staining. Results: There were no significantly statistical differences in the body weight,mass and organ/body coefficients of testis and epididymis,sperm quantity,sperm motility rate among four groups of P、F1、F2 male rats （P>0.05）. There were no significantly statistical differences in sperm malformation rate between the high-dose group and the control group （P>0.05）. There was no significant change in testis and epididymis of P,F1 and F2 male rats. Conclusion Dendrobium officinale flowers did not show obviously adverse effects on testivular tissue morphology and sperm quality in parent and offsping rats.

Objective: To evaluate the safety of a herbal pilatory for external use. Methods : An acute eye irritation test were employed to detect the eye irritation of the herbal pilatory;a skin irritation test,a skin sensitization test and a skin phototoxicity test were employed to detect the dermal toxicity;Salmonella typhimurium reverse mutation assay（Ames test）and chromosome aberration test in CHL cells were employed to detect the effects of the pilatory on gene mutation and chromosome aberration in prokaryotic and eukaryotic cells. Results: When the eyes of rabbits exposed to the pilatory without rinse during the first 24 hours,the conjunctiva showed congestion and edema with the highest score of 2,corneal opacity was observed with the highest score of 1;however,these symptoms returned to normal within 72 hours,with the score reduced to 0. No irritation to the skin of rabits was found after exposed to the pilatory for fourteen days,no skin sensitization was introduced by Buehler test and no skin phototoxicity on guinea pigs was detected. There was no abnormal growth of reverse mutation colonies induced by the pilatory under S9 acitivation or not. There was no statistically significant rise of chromosome aberration rate in the exposed CHL cells compared to the control groups（P>0.05）. Conclusion Under the condition,the herbal pilatory showed mild and reversible irritation to eyes,while no dermal toxicity and genetic toxicity were observed. The safety of the herbal pilatory for external use is acceptable.

Objective:To analyze the clinical characteristics and prognosis of patients with primary aldosteronism (PA) associated with subclinical Cushing syndrome (SCS).Methods:This retrospective cohort study was conducted at the First Affiliated Hospital of Chongqing Medical University in China. Patients with PA were included between January 2014 and December 2022. According to the results of 1-mg overnight dexamethasone suppression test, the patients were divided into the PA group and PA associated with SCS (PA/SCS) group. The demographic information, hormone levels, and follow-up results were analyzed. Independent sample t-test, chi-square test and Mann-Whitney U test were used for data comparison. Results:A total of 489 PA patients were enrolled in this study, of which 109 had PA/SCS (22.3%). Patients with SCS were on average older (54.4±10.7 vs . 47.4±11.0, P<0.001); had a larger proportion of women (69.7%, 76/109 vs . 57.4%, 218/380; P=0.020); and a longer duration of hypertension [96 (36, 180) vs . 60 (12, 120) months, P=0.001] than patients without SCS. There were 215 and 51 patients in the PA group and PA/SCS group, who completed adrenalectomy and follow-up, respectively. The remission rate of autonomous cortisol secretion in the PA/SCS group was 85.3% (29/34). There was no significant difference in the remission rate of autonomous aldosterone secretion among patients between the PA/SCS and PA group (94.1%, 48/51 vs. 94.4%, 203/215; P=1.000), while the clinical remission rate in the PA/SCS group was lower than that in the PA group (39.2%, 20/51 vs. 61.9%, 133/215; P=0.003). Conclusions:SCS is common in PA patients (22.3%), and the clinical remission rate is low. Screening using the 1-mg overnight dexamethasone suppression test is recommended for all patients with PA.