Trophozoites of Giardia lamblia were axenically cultivated with modified TYI-S-33 medium contained 500 ?g/ml metronidazole(12h LC50).The morphology of drug-treated trophozoites was observed with light and electron micro-scopes at 2,4,8,12 h respectively.The light microscopy revealed that the trophozoites treated with MTZ showed swollen,detached from the wall of the culture tube,and were with vacuoles in the cytoplasm.Movement of the flagella become slowly or stopped.Electronic microscopy showed that the trophozoites were swollen and deformed;lots of vacuoles were seen in the cytoplasm;the contents of cytoplasm were depleted and the nuclei deformed.This study indicated that MTZ has injured the morphology of G.lamblia.

OBJECTIVE:To provide reference for the approval of Chinese medicines’patent application in the United States. METHODS:The requirements of patent eligibility in the United States were interpreted;two important patent cases(“Mayo case”and“Myriad case”)were analyzed in recent years,and the effects of“patent eligibility guideline”on Chinese medicines’patent ap-plication in the United States were also analyzed;the suggestions were put forward according to the situation of Chinese medicines’ patent application. RESULTS & CONCLUSIONS:Patent eligibility required that patent protection objects should be included in ob-ject range which could be vested patent right stated in patent law. The analysis of“Mayo case”“Myriad case”and patent eligibility guideline indicated that if related Chinese medicines’patent was to obtain patent approval in the United States,notable difference between the medicine and natural products as well as the order of nature must be clearly stated;technical attributes should be em-phasized,and different patent application ideas were adopted for different types of invention in order to guarantee patent eligibility. At present,small number of Chinese medicine’s patent in the United States come from China,and relevant enterprises should liber-ate themselves from the misperception that Chinese medicines do not hold patent eligibility. Hence,it is suggest that the first claim in the patent should not be too definite in Chinese medicines’patent application,afford more comprehensive application,strength-en patent protection of classic Chinese medicine recipe and stress patent eligibility.

Objective To evaluate prenatal diagnosis value of echocardiography in pathological types,differential diagnosis and accompanied malformations of fetal persistent truncus arteriosus(PTA).MethodsTwenty-four cases of PTA selected from 1 392 cases were analysed,who were definitely diagnosed to be suffered from cardiovascular malformation by fetal echocardiography.The ultrasound findings,pathological results and followed up were analysed.According to Van Praagh classification,the type IV PTA was excluded in this study which was classified into pulmonary artery atresia.Results The total PTA were 24 cases,in which 10 cases of A1 type,3 cases of A2 type,9 cases of A3 type,and 2 cases of A4 type.Nine cases of PTA accompanied other cardiac anomalies,and 1 case of PTA accompanied both cardiac anomalies and extracardial malformations.Two PTA cases were born,one was A1 type underwent surgical intervention,and the other was died due to multiple organ-failure.Fourteen PTA cases were termination and 7 cases were confirmed by pathology.Seven women pregnant again,of which 5 cases were born while only one was diagnosed atrial septal defect after birth,2 pregnant women were still during follow-up.Eight PTA cases follow-up were lost.Conclusions A1 type and A3 type of PTA have high incidence in fetus.Accompanied cardiac anomalies is certainly related to different types.Combination of multiple ultrosund techniques can diagnose PTA prenatally,make accurate classification and detect accompanying malformations,which is of great significance to offer proper pregnancy counselling and postpartum treatment.

BACKGROUND:Apoptosis of islet cel s is closely related to the long-term hyperglycemia-and hyperlipemia-induced injuries. OBJECTIVE:To observe the effect of Shizidaiping formula on the apoptosis and insulin secretion in MIN6 cel s under the high glucose and lipid environment, and to explore the protective effect of Shizidaiping formula and the related apoptosis mechanism. METHODS:MIN6 cel s were divided into normal, model, melbine, low-, medium-and high-dose Shizidaiping formula groups. The cel activity was examined by cel counting kit-8, the insulin secretion was measured by ELISA, the rate of apoptosis was measured by Annexin V-FITC&PI and the expression levels of MEK1/2, ERK1/2 and p-ERK1/2 were examined by western blot assay. RESULTS AND CONCLUSION:Shizidaiping formula significantly improved MIN6 cel activity under high glucose and lipid condition (P<0.05), decreased early cel apoptosis, increased the level of insulin stimulated by low glucose in cel supernatant (P<0.05), and improved the expression levels of MEK1/2, ERK1/2 and p-ERK1/2 (P<0.05). These results suggest that Shizidaiping formula can protect islet cel s from hyperglycemia and hyperlipemia damage by improving the activity of MIN6 cel s, reducing the insulin secretion and inhibiting the apoptosis of pancreaticβcel s in MIN6 cel s.

The purpose of this study was to investigate the role of miR-4281 in immune thrombocytopenic purpura(ITP)and its underlying mechanism.Differentially expressed miRNAs in ITP were searched by using GEO database.Newly diagnosed ITP patients,treatment-effective ITP patients and healthy subjects were collected.Venous blood was obtained and peripheral blood mononuclear cells(PBMCs)and CD4+T cells were separated.Real-time quantitative PCR was used to detect the expression of miR-4281 and other factors in cells.Flow cytometry was conducted to analyze the rate of CD4+CD25+FOXP3+Tregs and Th17 cells.Pearson correlation coefficient method was used to test the relationship between miR-4281 expression level and various factors.Bioinformatics analysis revealed an abnormal expression of miR-4281 in ITP.Compared with healthy subjects and treatment-effective ITP patients,the expression level of miR-4281 in newly diagnosed ITP was decreased.The expression level of miR-4281 in PBMCs of ITP patients was positively correlated with Tregs content and Treg/Th17,while negatively correlated with Th17 cell content.miR-4281 can promote the differentiation of Tregs cells and inhibit the differentiation of Th17.Furthermore,the expression level of miR-4281 is positively correlated with the expression level of FOXP3.Taken together,the expression level of miR-4281 is reduced in ITP.This decrease can inhibit the expression of FOXP3,cause Treg/Th17 immune imbalance,and promote the progression of ITP.

Objective:To understand the current situation and influencing factors of depression among residents in Chongqing, and to explore the relationship between depression and administration satisfaction of the standardized residency training.Methods:Stratified random cluster sampling was used to select 372 residents currently participating the standardized residency training in Chongqing. The patient health questionnaire (PHQ-9) and administration satisfaction of residency training questionnaire were used to conduct a cross-sectional survey on the depression status and satisfaction of residents in Chongqing. SPSS 21.0 software package was used for partial correlation analysis and binary logistic regression analysis to explore the relationship between depression and administration satisfaction of the standardized residency training.Results:Among the 372 residents, 218 had mild depression and above (58.60%). There was a significant difference in the incidence of depression among residents of different ages, educational background, marital status, working hours and administration satisfaction of residency training ( χ2 = 23.02, 17.35, 12.24, 21.70, 95.20; P = 0.028, 0.027, 0.016, 0.041, <0.001 respectively). Partial correlation analysis showed that there was a negative correlation between the total score of depression and the five aspects of the administration satisfaction (total score, instructors, training base, managers and training income) ( r=-0.52, -0.45, -0.47, -0.45, -0.40, all P<0.01). In the binary logistic regression analysis of depression and satisfaction,the satisfaction had a negative impact on depression, and its B value was -1.71, and OR value was 0.18, P<0.001. Conclusion:The incidence of depression among residents in Chongqing is high. Age, educational background, marital status, working hours and administration satisfaction are related to the degree of depression. And the higher the degree of depression, the lower the administration satisfaction of residents. Relevant departments should pay attention to the mental health of residents and improve the quality of standardized residency training.

By analyzing the of genetic testing data of patients with renal polycystic kidney disease and their relatives, this study aims to identify unreported novel gene mutation sites associated with autosomal dominant polycystic kidney disease (ADPKD). Structural prediction software was employed to investigate protein structural changes before and after mutations, explore genotype-phenotype correlations, and enrich the ADPKD gene database. In this single-center retrospective study, patients with multiple renal cysts diagnosed from January 2019 to February 2023 at the Zhong Da Hospital Southeast University were included. Genetic and clinical data of patients and their families were collected. Unreported novel gene mutation sites associated with ADPKD were identified. The AlphaFold v2.3.1 software was used to predict protein structures. Changes in protein structure before and after mutations were compared to explore genotype-phenotype correlations and enrich the ADPKD gene database. Twelve mutated genes associated with renal cysts were detected in 52 families. Nineteen novel gene mutation sites associated with ADPKD were identified, including 17 mutations in the PKD1 gene (one splicing mutation, seven frameshift mutations, four nonsense mutations, one whole-codon insertion, and four missense mutations); one ALG9 missense mutation; and one chromosomal structural variation. Truncating mutations in the PKD1 gene were correlated with a more severe clinical phenotype, while non-truncating mutations were associated with greater clinical heterogeneity. Numerous novel gene mutation sites associated with ADPKD remain unreported. Therefore, it is essential to analyze the pathogenicity of these novel mutation sites, establish genotype-phenotype correlations, and enrich the ADPKD gene database.

Objective:To investigate the effect of hourly ambient temperature exposure on emergency calls in Xuchang city and Zhengzhou city.Methods:The hourly meteorological data, air pollution data and emergency calls of Xuchang city and Zhengzhou city were collected from January 1, 2017 to October 31, 2019. A distributed lag non-linear model was used to calculate the excess relative risk ( ERR). The lag effect and cumulative effect of extreme temperature exposure on emergency calls were evaluated. Results:The relationship between hourly temperature exposure and emergency calls was a U-shaped curve. In Xuchang city and Zhengzhou city, both low and high temperatures would increase the number of hourly emergency calls. The earliest effect of low temperature occurred at a lag of 22 h and 52 h, with ERR values (95% CI) about 0.20% (0.00%, 0.39%) and 0.11% (0.00%, 0.22%), respectively. The earliest effect of high temperature occurred at a lag of 0 h with ERR values about 1.59% (1.09%, 2.09%) and 1.45% (1.22%, 1.69%), respectively. High temperature had the greatest impact on the number of emergency calls of cardiovascular disease at a lag of 4-8 h. The cumulative ERR values (95% CI) of the two cities were 8.70% (4.98%, 12.75%) and 3.89% (2.61%, 5.22%), respectively. Conclusion:High temperature exposure could increase the number of emergency calls within a few hours, while the effect of low temperature would not occur until 22 hours later.

Objective:To evaluate the effect of electroacupuncture on liver regeneration after partial hepatectomy in mice and the role of the Notch signaling pathway.Methods:Thirty-six SPF healthy male C57BL/6 mice, aged 6 weeks, weighing 20-22 g, were divided into 6 groups ( n=6 each) using a random number table method: sham operation group (group S), partial hepatectomy group (group PH), non-acupoint electroacupuncture+ partial hepatectomy group (group NPH), partial hepatectomy+ Fli-06 group (group PH+ F), acupoint electroacupuncture+ partial hepatectomy group (group EPH), and acupoint electroacupuncture+ partial hepatectomy+ Fli-06 group (group EPH+ F). All the mice except for group S underwent partial hepatectomy. Fli-06 4.8 mg/kg was intraperitoneally injected starting from 2 days before surgery, once a day, until the mice were sacrificed in group PH+ F and group EPH+ F, while the equal volume of 0.9% sodium chloride solution was injected in the other groups. In EPH group, electroacupuncture of bilateral " Zusanli" acupoints lasting for 15 min was performed using continuous waves with a frequency of 2 Hz and an intensity of 1 mA once a day starting from the time point immediately after surgery for 3 consecutive days. Mice were anesthetized at day 2 after partial hepatectomy, and blood samples were taken from the eyeball for determination of the serum alanine transaminase (ALT) and aspartate aminotransferase (AST) concentrations (using a fully automated biochemical analyzer) and concentrations of serum epidermal growth factor (EGF) and hepatocyte growth factor (HGF) (by enzyme-linked immunosorbent assay). The mice were subsequently sacrificed and liver tissues were taken for calculation of the liver mass to body mass ratio and for determination of the expression of liver proliferation marker Ki-67 (by immunohistochemical staining), proliferating cell nuclear antigen (PCNA), cyclin D1 (CCND1), Notch Intracellular Domain (NICD), and hypoxia-inducible factor-1alpha (HIF-1α) (using Western blot) and Notch1, jagged canonical Notch ligand 1 (Jagged1) and hairy and enhancer of split 1 (Hes1) mRNA (by real-time polymerase chain reaction). Results:Compared with group S, the serum ALT, AST, EGF and HGF concentrations were significantly increased, and the expression of hepatic Notch1, Jagged1 and Hes1 mRNA and Ki-67, PCNA, CCND1 and NICD was up-regulated in group PH ( P<0.05 or 0.01). Compared with group PH, the liver mass to body mass ratio and serum EGF and HGF concentrations were significantly increased, the serum ALT and AST concentrations were decreased, and the expression of hepatic Notch1, Jagged1, Hes1 mRNA and Ki-67, PCNA, CCND1, NICD and HIF-1α was up-regulated in group EPH, and the liver mass to body mass ratio and the serum HGF concentrations were significantly decreased, the serum ALT and AST concentrations were increased, and the expression of hepatic Jagged1 and Hes1 mRNA and Ki-67, PCNA, CCND1, NICD, and HIF-1α was down-regulated in group PH+ F ( P<0.05 or 0.01). Compared with group EPH, the liver mass to body mass ratio and serum EGF and HGF concentrations were significantly decreased, the serum ALT and AST concentrations were increased, and the expression of hepatic Notch1, Jagged1, Hes1 mRNA and Ki-67, PCNA, CCND1, NICD and HIF-1α was down-regulated in group EPH+ F ( P<0.01). Conclusions:Electroacupuncture at Zusanli acupoint promotes liver regeneration after partial hepatectomy in mice, and the mechanism may be related to the activation of the Notch signaling pathway.

Objective:To investigate the effect of hourly ambient temperature exposure on emergency calls in Xuchang city and Zhengzhou city.Methods:The hourly meteorological data, air pollution data and emergency calls of Xuchang city and Zhengzhou city were collected from January 1, 2017 to October 31, 2019. A distributed lag non-linear model was used to calculate the excess relative risk ( ERR). The lag effect and cumulative effect of extreme temperature exposure on emergency calls were evaluated. Results:The relationship between hourly temperature exposure and emergency calls was a U-shaped curve. In Xuchang city and Zhengzhou city, both low and high temperatures would increase the number of hourly emergency calls. The earliest effect of low temperature occurred at a lag of 22 h and 52 h, with ERR values (95% CI) about 0.20% (0.00%, 0.39%) and 0.11% (0.00%, 0.22%), respectively. The earliest effect of high temperature occurred at a lag of 0 h with ERR values about 1.59% (1.09%, 2.09%) and 1.45% (1.22%, 1.69%), respectively. High temperature had the greatest impact on the number of emergency calls of cardiovascular disease at a lag of 4-8 h. The cumulative ERR values (95% CI) of the two cities were 8.70% (4.98%, 12.75%) and 3.89% (2.61%, 5.22%), respectively. Conclusion:High temperature exposure could increase the number of emergency calls within a few hours, while the effect of low temperature would not occur until 22 hours later.