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@#Abstract: Transfection of Plasmodium falciparum is helpful to study the function of its genes, such as drug resistance. However, transgenic manipulation has been very challenging, mainly due to the high A/T base sequence structure (A+T content of about 82%) and low transfection efficiency of the Plasmodium genome. Electroporation-based transfection of Plasmodium falciparum has been successfully applied in the study of certain genes, and electroporation by preloading is currently the preferred method for introducing foreign DNA into Plasmodium falciparum. The site-directed editing of Plasmodium genes mostly adopts the method of two-plasmid transfection. It is generally believed that successful transfection of Plasmodium requires a large amount of high-purity plasmid DNA and an accurate transfection system. In addition to the evaluation of the current commonly used electrotransfection methods, this paper also introduces a new transfection method, namely lyse-reseal erythrocytes for transfection (LyRET). This paper also review the role of factors such as plasmid DNA concentration, the use of transfection reagents, the setting of transfection parameters, the addition of fresh red blood cells, and the markers of successful transfection in improving the success rate and efficiency of Plasmodium transfection, in the hope of providing a reference for study in this field.
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Objective:To explore the adjunctive diagnostic value of transcranial sonography (TCS) combined with olfactory test in early Parkinson′s disease (PD) and the clinical value of both in the cognitive function of PD patients.Methods:TCS and olfactory test were performed in 157 early PD patients(PD group) and 157 healthy controls(control group) in the Second Affiliated Hospital of Soochow University from January 2018 to January 2022. The differences in clinical characteristics, TCS, and olfactory test results between the two groups were analyzed. The values of TCS, olfactory test, and their combination in diagnosing early PD were evaluated using clinical diagnosis as the gold standard. The correlations of the midbrain area, the midbrain substantia nigra hyperechoic area, and the third ventricle width in TCS examination with the cognitive score were analyzed in the PD group. According to the olfactory test scores, 157 patients with early PD were divided into two groups: 110 cases of PD with olfactory dysfunction (PD-OD) and 47 cases of PD without olfactory dysfunction (PD-NOD). The differences in clinical scores and TCS results between the two groups were compared.Results:The midbrain substantia nigra hyperechoic area, substantia nigra hyperechoic positivity rate, third ventricle width, and olfactory dysfunction rate were higher in the PD group compared to the control group, while the midbrain area and olfactory test scores were lower than those in the control group (all P<0.001). The sensitivity and the coincidence rate of TCS combined with the olfactory test for early PD diagnosis (90.0%, 77.1%) were higher than those of TCS alone (60.0%, 71.3%) and olfactory test alone (70.1%, 72.3%), but the specificity (63.7%) was lower than that of both alone (82.8% for TCS and 75.2% for olfactory test), (all P<0.001). MoCA score, visual space and executive ability, memory, attention, and language were positively correlated with the area of the midbrain ( rs=0.38, 0.32, 0.27, 0.25, 0.23; all P<0.05) and negatively correlated with the width of the third ventricle ( rs=-0.39, -0.22, -0.39, -0.22, -0.32; all P<0.05), and orientation was negatively correlated only with the width of the third ventricle ( rs=-0.24, P<0.05). The MoCA score of PD-OD group[22(18, 25)] was lower than that of PD-NOD group[24(20, 26)]( P=0.040). Conclusions:The combination of TCS and olfactory test can enhance the sensitivity and diagnostic agreement rate for early PD diagnosis, providing some auxiliary value. The cognitive function of PD patients is positively correlated with the midbrain area and negatively correlated with the width of the third ventricle. The cognitive function of PD patients with olfactory dysfunction is lower than that of PD patients without olfactory dysfunction. TCS and olfactory test may help assess cognitive function in PD patients.
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Objective:To explore the pathological mechanism of SN hyperechogenicity by investigating the characteristics of substantia nigra (SN) hyperechogenicity on transcranial sonography (TCS) and serum iron metabolism parameters in the postural instability gait difficulty and tremor dominant subtypes of Parkinson′s disease (PD), and the correlation between them.Methods:A total of 155 PD patients recruited in Parkinson′s Disease Specialty in the Second Affiliated Hospital of Soochow University from January 2019 to December 2021 were divided into postural instability gait difficulty group( n=95) and tremor dominant group( n=60). Meanwhile, 49 healthy gender- and age-matched healthy individuals who sought for physical examination during the same period were included as the control group. All subjects underwent TCS and blood test, and the echo of SN between the postural instability gait difficulty group and tremor dominant group, serum iron metabolism parameters among the three groups were compared. The postural instability gait difficulty group and tremor dominant group were subdivided into with SN hyperechogenicity (SN+ )subgroup and without SN hyperechogenicity (SN-) subgroup respectively according to TCS results, and the differences in serum iron metabolism parameters between the subgroups were further compared. The association between SN hyperechogenicity and serum iron metabolism parameters of the postural instability gait difficulty group and tremor dominant group were further analyzed. Results:The total area of bilateral SN+ , the area of SN+ on the larger side, and the ratio of the total area of SN+ to the midbrain area (S/M) in postural instability gait difficulty group were larger than those in tremor dominant group (all P<0.001). The value of serum ceruloplasmin and transferrin in both postural instability gait difficulty group and tremor dominant group were lower than those in control group (all P<0.001), and compared with tremor dominant group and control group, the postural instability gait difficulty group had lower serum ferritin(all P<0.01). In both postural instability gait difficulty group and tremor dominant group, serum ceruloplasmin in SN+ subgroup was lower than that in SN-subgroup ( P=0.001, 0.032). Moreover, there was a negative correlation between serum transferrin and the area of SN hyperechogenicity in two subgroups(postural instability gait difficulty group: rs=-0.454, P<0.001; tremor dominant group: rs=-0.494, P<0.001). Conclusions:Compared with the tremor dominant patients, the postural instability gait difficulty patients have larger area of SN hyperechogenicity and lower serum ferritin level. The area of SN hyperechogenicity is significantly negatively correlated with serum transferrin level, indicating that the production of this imaging characteristics is related to iron metabolism.