Objective To develop a droplet digital PCR method for detection of baculovirus(BV), optimize the annealing temperature, primer and probe concentrations, and verify the method.Methods According to the conserved sequence characteristics of GP64 gene, specific primers and TaqMan probes were designed to develop a droplet digital PCR detection method for BV. The annealing temperature, primer concentration, and probe concentration of the system were optimized, and the specificity, reproducibility, sensitivity and linearity of the method were verified. The 24 samples from GMP pilot workshop were detected by the droplet digital PCR, and the results were compared with those detected by plaque method.Results A droplet digital PCR method targeting BV was developed. The optimized annealing temperature, primer and probe concentrations were 58. 0 ℃, 400 nmol/L, and 200 nmol/L, respectively. Positive droplets were detected only in the reaction with BV nucleic acid as template, which showed that the method had good specificity. The CVs of intra-batch reproducibility ranged from 2. 78% to 7. 45%, and those of inter-batch reproducibility ranged from 2. 28% to 7. 05%, both less than10%, indicating that the method had good reproducibility. The minimum detection limit was 3. 06 copies/μL, which was about 100 times higher than that of the ordinary PCR, showing the high sensitivity of the method. The results of 24 samples were close to those of plaque method, and the correlation coefficient(r)was 0. 913.Conclusion An optimized droplet digital PCR assay for the detection of BV has been successfully developed, which has good specificity, reproduci-bility, sensitivity,and rapidness, and can be used for the rapid detection of BV in production.

Objective To investigate the therapeutic effect of ipratropium bromide on bronchial spasm of patients with asthma and chronic obstructive pulmonary disease (COPD).Methods 174 patients with asthma and COPD were selected and divided into two groups, 87 cases in the control group treated with budesonide and formoterol fumarate powder for inhalation , 87 cases in the experimental group received ipratropium bromide on the basis of the control group, pulmonary ventilation function, airway resistance and cytokine levels in induced sputum, the clinical effect and incidence of adverse reactions were compared after the treatment.Results The effective rate in the control group(81.61%)was lower than the experimental group (93.11%), with significant difference (P<0.05); compared with the control group, levels of peak expiratory flow (PEF), maximum midexpiratory flow (MMEF), forced expiratory volume in one second to forced vital capacity ratio (FEV1%), forced vital capacity (FVC) were higher in the experimental group after treatment, levels of airway resistance (Raw), Raw% were lower, level of specific airway conductance (Gsp) was higher after treatment, induced sputum levels of vascular endothelial growth factor (VEGF), intercellular adhesion molecular-1 (ICAM-1), interleukin-13 (IL-13), IL-17 were lower after treatment, with significant difference (P<0.05);there was no significant difference in the incidence of adverse reactions between the two groups.Conclusion The clinical effect of ipratropium bromide in the treatment of asthma and COPD was exactly , ipratropium bromide can significantly improve the pulmonary ventilation function in patients, relieve bronchial spasm, reduce airway resistance, inhibit airway inflammation, and the safety is higher.

OBJECTIVETo reveal the flowering characteristics of Panax stipuleanatus and provide theoretical basis for distant hybridization between P. stipuleanatus and P. notoginseng.

METHODDuring the blossom of P. stipuleanatus, we observed and investigated blooming phenophase, growth dynamics of inflorescence, pollination and seed setting of its population and the flowering process and its period, the regularity of flowering and pollinating of the floret. Statistic analyses were carried out.

RESULT AND CONCLUSIONThe population florescence of P. stipuleanatus was about 60 d. The average florets quality of umbel was 44-47. The average natural pollination rates were 73.32%-95.39%. The average seed setting rates was 35.65%-51.76%. The highest growth periods of inflorescence are from March 25 to April 4. The diameter, the height and the length of its inflorescence in 10 d increased 44.65%, 42.19% and 106.25%, respectively. The whole stereotype period was the May beginning. The time that the floret from petal opening to withering it generally needed 36-48 h, 60 percent of the floret finished auther pollinating during the same day and 40 percent until the next day. The flowering and pollinating peak periods were 14:00-15:00, the flowering numbers was 28.48% of the total flowering amount and the pollinating numbers were 38.63% of the total pollinating amount. The high temperature (20-30 degrees C) and the low humidity (RH < 60%) were beneficial to flowering and pollinating.

Flowers ; growth & development ; physiology ; Inflorescence ; growth & development ; physiology ; Panax ; growth & development ; physiology ; Pollination ; Seeds ; physiology

lusions WI parameters reflect both cardiac and vascular changes of hyperthyroid patients. Mid-systolic hemodynamic turbulence appears in a portion of hyperthyroid patients.

Characteristics of fructification and seed ripening of Panax stipuleanatus were studied in this paper with a view to better understand the species' biological characteristics and promote cultivation. Dynamic of fructification and types of fruits were observed in a period of 80 days. The seeds' biological characteristics were evaluated by observing embryo's vitality and changes in form in a period of 225 d. The findings and conclusions were as follows: A plant produces an average of (38.7 +/- 6.5) seeds; Seeds become fully mature 80 days after setting fruit, and fruits grow more rapidly in 0-40 days with an increase of 1 500.83% in fresh weight. Three types of fruits were found, that is mono-seed type, bi-seeds type and triple-seeds type, and the proportion is 9.53%, 58.66%, 1.81%, respectively. The seeds are (5.16-6. 52) mm in length and (3.12-4.95) mm in width. The weight of 1000 seeds is (0.648 +/- 0.014) g with a moisture content of (55.23 +/- 0.49)%. The after-ripening of embryo in Panax stipuleanatus is morphological form, embryos develop 75 days after the seeds' detaching from mother plant and become mature in 165-180 days with the ending of the process of after-ripening. The vitality of embryo reaches 70% , and the length of embryo is (3.217 +/- 0.406) mm and the width (0.872 +/- 0.165) mm. The seeds will germinate in 195 d.
Fruit ; growth & development ; Germination ; physiology ; Kinetin ; biosynthesis ; metabolism ; physiology ; Panax ; chemistry ; physiology ; Physiological Phenomena ; Seeds ; physiology

lerosis of the lower extremity artery in patients with type 2 MD.

This study examined the wave intensity (WI) of the carotid artery in patients with hyperthyroid in order to assess the hemodynamic changes of hyperthyroid patients. A total of 86 hyperthyroid patients without cardiac morphological changes and arrhythmia, and 80 healthy control subjects were enrolled in the study. Right common carotid artery (RCCA) was selected for ultrasonic imaging to obtain WI indices, including amplitude of the peak during early systole (W1), amplitude of the peak during late systole (W2), area of the negative wave during mid-systole (NA), interval between R wave of electrocardiogram and W1 (R-1st), interval between W1 and W2 (1st-2nd). The levels of serum thyroid hormones, consisting of free triiodothyronine (FT3), free thyroxin (FT4) and thyroid stimulating hormone (TSH), were measured in hyperthyroid patients. Echocardiographic indices including left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS) were determined in each subject. The results showed that the W1, W2, NA, and (1st-2nd×HR) in hyperthyroid patients were significantly higher than those in healthy controls. There was no significant difference in LVEF and LVFS between the two groups. FT3 was correlated with W1, W2, NA, (1st-2nd×HR), pulse pressure (PP) and heart rate (HR) in hyperthyroid patients. Several abnormal waves on WI curves were present in 19 hyperthyroid patients during mid-systole. It was concluded that WI technique may prove a real-time, noninvasive, sensitive and convenient tool for assessing the cardiac function and hemodynamic alterations in hyperthyroid patients.

Objective:To investigate the role of rapamycin target protein (mTOR) in lanthanum-induced injury of cerebral cortical neurons in offspring rats, and the effect on brain development, learning and memory ability of offspring rats.Methods:Thirty-two adult female and 32 male Wistar rats, were randomly divided into 4 groups according to their body weight, with 16 rats in each group (half female and half male). Female rats were fed with different amounts of lanthanum chloride[0.0 (control), 2.5, 5.0, 10.0 g/L], while male rats drank normal water. Female and male rats were mated in cages at a ratio of 1∶1. Female rats began to be exposed to lanthanum from pregnancy, while their offspring were exposed to lanthanum until 4 weeks after weaning. Morris water maze experiment was carried out in the 4 groups of offspring rats, and the effects of lanthanum on learning and memory were observed by space exploration. The cerebral cortex of offspring rats was taken, and the amount of Nissl body was observed under microscope after Nissl staining. The expression of mTOR mRNA in offspring rats cerebral cortex nerve cells was measured by real-time quantitative PCR. Western blotting was used to detect the protein content of p-mTOR in offspring rats cortical neurons.Results:Compared with the control group, the body weight of offspring rats exposed to lanthanum at 2.5, 5.0 and 10.0 g/L was significantly decreased [(121.75 ± 11.20), (110.00 ± 11.59), (98.88 ± 7.95) and (85.63 ± 7.25) g, P < 0.05], and the brain tissue coefficient and cortical coefficient were significantly increased [(1.43 ± 0.10)%, (1.56 ± 0.18)%, (1.66 ± 0.14)%, (1.89 ± 0.16)%; (0.86 ± 0.08)%, (0.94 ± 0.08)%, (1.01 ± 0.07)%, (1.08 ± 0.09)%, P < 0.05]. The brain weight [(1.63 ± 0.05), (1.61 ± 0.03) g] of 5.0 and 10.0 g/L lanthanum-exposed groups were significantly lower than those in the control group and 2.5 g/L lanthanum-exposed group [(1.73 ± 0.06), (1.70 ± 0.06) g, P < 0.05]. Compared with the control group (53.25 ± 9.93), the amounts of Nissl body in cerebral cortical neurons in different lanthanum-exposed groups (36.13 ± 3.98, 27.50 ± 5.21, 13.63 ± 5.93) were significantly decreased ( P < 0.05). The results of space exploration experiment showed that compared with the control group [(5.75 ± 1.98) times, (10.69 ± 2.96) s, (3.75 ± 1.28) times], the times of entering the target quadrant [(3.63 ± 1.41) times] and the stay time in the target quadrant [(5.12 ± 2.09) s] in 10.0 g/L lanthanum-exposed group were significantly reduced ( P < 0.05), and the times of entering the platform [(1.88 ± 0.84), (1.13 ± 1.12) times] in 5.0 and 10.0 g/L lanthanum-exposed groups were significantly reduced ( P < 0.05). There were significant differences in mTOR mRNA (1.00 ± 0.28, 0.74 ± 0.19, 0.58 ± 0.13, 0.45 ± 0.29) and p-mTOR protein expression levels (0.69 ± 0.07, 0.33 ± 0.06, 0.30 ± 0.04, 0.17 ± 0.03) in cortical tissues ( F = 8.33, 139.12, P < 0.05). Conclusion:Lanthanum exposure can damage cortical neurons, affect the brain development of offspring rats, reduce the expression of mTOR mRNA and p-mTOR protein in the brain of offspring rats, reduce the ability of space exploration and observation, resulting in the decline of learning and memory ability of offspring rats.

This study examined the wave intensity (WI) of the carotid artery in patients with hyperthyroid in order to assess the hemodynamic changes of hyperthyroid patients. A total of 86 hyperthyroid patients without cardiac morphological changes and arrhythmia, and 80 healthy control subjects were enrolled in the study. Right common carotid artery (RCCA) was selected for ultrasonic imaging to obtain WI indices, including amplitude of the peak during early systole (W1), amplitude of the peak during late systole (W2), area of the negative wave during mid-systole (NA), interval between R wave of electrocardiogram and W1 (R-1 st), interval between W1 and W2 (1 st-2nd). The levels of serum thyroid hormones, consisting of free triiodothyronine (FT3), free thyroxin (FT4) and thyroid stimulating hormone (TSH), were measured in hyperthyroid patients. Echocardiographic indices including left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS) were determined in each subject. The results showed that the WI, W2, NA, and (lst-2nd×HR) in hyperthyroid patients were significantly higher than those in healthy controls. There was no significant difference in LVEF and LVFS between the two groups. FT3 was correlated with WI, W2, NA,(lst-2nd×HR), pulse pressure (PP) and heart rate (HR) in hyperthyroid patients. Several abnormal waves on WI curves were present in 19 hyperthyroid patients during mid-systole. It was concluded that WI technique may prove a real-time, noninvasive, sensitive and convenient tool for assessing the cardiac function and hemodynamic alterations in hyperthyroid patients.

Objective To comprehensively analyze the reported preparation methods for animal models of perimenopausal syndrome (PS), to compare the advantages and disadvantages of various preparation elements and detection indexes, so as to provide useful references for the optimization of the relevant animal models as well as the standardization of their application in the efficacy evaluation of new drugs.MethodsIn this paper, literature research methods were applied using "perimenopausal syndrome" as the subject term. The publication period of the literature was limited to January 2016 to February 2023. Relevant literature on the preparation of PS animal models was retrieved from databases such as China National Knowledge Infrastructure, Wanfang database, and PubMed. After screening the experimental literature that met the inclusion and exclusion criteria, detailed information on experimental animal strains, modeling methods, duration of drug administration, positive drugs, detection indexes and other relevant information were collected. After the above information was standardized, the PS animal model database was established using Excel 2010 software. The model preparation elements and evaluation indexes were summarized systematically, and the statistical results were processed and analyzed using Excel 2010 software.Results A total of 247 articles were screened. SD rats (164 times, 65.86%) and Wistar rats (35 times, 14.06%) were often used to prepare PS animal models. Bilateral ovariectomy (139 times, 53.87%) and natural aging (43 times, 16.80%) were chosen as modeling methods. The ages of rats used for modeling ranged from 7 weeks to 18 months, with 3-month-old rats (22 times, 21.78%) being the most common. The detection indexes were comprehensively evaluated from multiple perspectives, including serum biochemistry, vaginal exfoliated cell smear, histomorphology, general observation, behavioral observation, and organ tissue protein immunoblotting. Western medical evaluation indexes were commonly used to test the successful preparation of models, with vaginal exfoliated cell smears being the most frequently used method (125 times, 85.04%). A model was considered successfully prepared when estrous cycle disorder or irregularity was observed. Some literature also determined modeling success by detecting a significant decrease in serum estradiol levels (5 times, 3.04%). Traditional Chinese medicine (TCM) syndrome evaluation often used a combination of Chinese and Western medical evaluation indexes for comprehensive evaluation, with researchers determining the TCM syndrome through vaginal exfoliated cell smears supplemented by general observation (3 times, 2.04%).Conclusion There are many methods for preparing PS animal models, but there are still significant differences in the selection of animal species, age, criteria for successful modeling, and TCM syndrome evaluation in the related literature.