The polymorphism and thermostability of nirmatrelvir, the main antiviral component of the oral COVID-19 treatment drug, were studied. Four polymorphs of nirmatrelvir were prepared by recrystallization methods. Among them, Form 1 and nirmatrelvir methyl tert-butyl ether solvate (Form 2) had been reported in the literature, while nirmatrelvir isobutyl acetate solvate (NMTW-IBAC) and nirmatrelvir ethyl acetate solvate (NMTW-EA) are two new solvates. The crystal structures were characterized by single-crystal X-ray diffraction, powder X-ray diffraction, thermogravimetric analysis and differential scanning calorimetry. The thermostability of polymorphism and crystalline transformation were also investigated by combining Hirshfeld surface analysis and interaction energy analysis. The results showed that nirmatrelvir Form 1 belongs orthorhombic crystal system with the space group P2₁2₁2₁ and one nirmatrelvir molecule included in the asymmetric unit, which has the same crystal structure as nirmatrelvir Form 4 reported in the literature. Owing to its larger thermal expansion, the differences in crystallographic parameters obtained at different temperatures were found between Form 1 and Form 4. Three solvates of nirmatrelvir belonged to the iso-structural with monoclinic crystal system and the space group P2₁, in which the asymmetric unit contains one nirmatrelvir molecule and one solvent molecule. The thermal analysis results showed that nirmatrelvir Form 1 was a solvent-free crystal form with the best thermal stability and the strongest intermolecular hydrogen bonding. Among the three solvates, NMTW-EA has the worst thermal stability and the weakest hydrogen bonding interaction between the nirmatrelvir molecule and the solvent molecule. The energy framework of nirmatrelvir solvates showed that the closer the arrangement between solvent and nirmatrelvir molecules, the greater the total interaction energy between solvent and nirmatrelvir molecules. The phase transition studies of the three solvates showed that NMTW-IBAC and NMTW-EA were transformed into amorphous after desolvation, respectively, while Form 2 undergoes oiling during desolvation. The research provides theoretical guidance for the analysis, identification and quality control of nirmatrelvir polymorphs.

The 1H-NMR fingerprints of three different species tibetan medicine sea buckthorn were established by 1H-HMR metabolomics to find out different motablism which could provide a new method for the quality evaluation of sea buckthorn. The obtained free induction decay (FID) signal will be imported into MestReNova software and into divide segments. The data will be normalized and processed by principal component analysis and.partial least squares discriminant analysis to perform pattern recognition. The results showed that 25 metabolites belonging to different chemical types were detected from sea buckthorn,including flavonoids, triterpenoids, amino acids, carbohydrates, fatty acids, etc. PCA and PLS-DA analysis showed three different varietiest of sea buckthorn that can be clearly separated by the content of L-quebrachitol, malic acid and some unidentified sugars, which can be used as the differences metabolites of three species of sea buckthorn. 1H-NMR-based metabonomies method had a holistic characteristic with sample preparation and handling. The results of this study can offer an important reference for the species identification and quality control of sea buckthorn.
Hippophae ; metabolism ; Magnetic Resonance Spectroscopy ; methods ; Medicine, Tibetan Traditional ; Metabolomics

OBJECTIVEIn order to figure out how and to what degree the social and economic development and control strategy influencing the epidemics of tuberculosis and to provide reference for tuberculosis prevention and control in China.

METHODSBased on the data from the nationwide random surveys on tuberculosis in 1979, 1984/1985, 1990 and 2000 and the indexes on social and economic development of China, correlation coefficient was used to analyze the relationship of three factors including (1) the change of epidemic situation of tuberculosis from 1979 to 2000; (2) the level of social and economic development; (3) the implementation of Health V Project.

RESULTSThe prevalence rate of smear positive tuberculosis was significantly correlated to per capita net income of rural population, consumption level of city population, per capita GDP, density of population, and proportion of rural population. Among which the correlation with per capita net in come of rural population, consumption level of city population, per capita GDP, or density of population showed negative, correlation but the proportion of rural population showed positive. The range of GDP increase was similar in both areas with or without the implementation of Health V Project from 1990 to 2000 (77.2% and 77.8%). However, the ranges of the decline of prevalence rate were quite different (44.4% and 12.3%) in the two areas. In the western part of China, the range of GDP increase was similar in the areas with or without the implementation of Health V Project. However, the prevalence rate declined in the area that implementing the project but increased in other areas without the project.

CONCLUSIONThe level of social and economic development had influenced the prevalence rate of tuberculosis, but the implementation of tuberculosis control project played an important role in the reduction of tuberculosis prevalence rate from 1979 to 2000 in China.

China ; epidemiology ; Female ; Humans ; Income ; statistics & numerical data ; Male ; Prevalence ; Rural Population ; Socioeconomic Factors ; Tuberculosis ; epidemiology

Heart Neoplasms ; diagnostic imaging ; secondary ; surgery ; Heart Ventricles ; pathology ; Humans ; Lung Neoplasms ; pathology ; Male ; Middle Aged ; Radiography

OBJECTIVETo study the current status of geriatric tuberculosis (TB) and its impact on TB control program under the directly observed treatment short-course (DOTS) strategy in China.

METHODSUsing the prevalence information regarding the epidemiology of geriatric TB from the National Random Survey in 2000, a case-control study was carried out to analyze the case detection, treatment and management of geriatric TB patients between DOTS area and non-DOTS area.

RESULTSThe prevalence of sputum smear positive (SS+) in the age group of 65 or above was 440/100 000 which was 3.6 times of the average prevalence of SS+ of all age groups. Geriatric SS+ patients took up 28.8% of all the TB patients in 13 provinces with implementation of DOTS and 28.9% in 15 provinces without. The population of TB case in the age group 65 or above occupied 11.4% of all the newly registered SS+ case in 13 DOTS provinces from 1992 to 2000.

CONCLUSIONThe prevalence of geriatric SS+ was high. In both DOTS and non-DOTS areas, the proportion of geriatric SS+ was high but the registration rate of new SS+ was low among all the age groups. Both high prevalence and low case detection rate of geriatric TB became main issues to be under concern in the TB control strategy in China.

Age Factors ; Aged ; Aged, 80 and over ; Case-Control Studies ; China ; epidemiology ; Communicable Disease Control ; statistics & numerical data ; Directly Observed Therapy ; statistics & numerical data ; Female ; Humans ; Male ; Middle Aged ; Outcome Assessment (Health Care) ; statistics & numerical data ; Prevalence ; Retrospective Studies ; Tuberculosis, Pulmonary ; epidemiology ; prevention & control ; World Health Organization

Two plasmid constructs, pcE2 and pcE3, containing 3' fragment of open reading frame 2 (ORF2,1163 bp) of hepatitis E virus (HEV) and full-length ORF3 (369 bp), were injected into bilateral tibialis of Swiss mice respectively，for three times (0, 2nd and 4th weeks) and observed the HEV IgG by ELISA. HEV IgG was induced after the injection of pcE2 or pcE3 or both, and the percentage of seraconversion was 100% after two weeks of the third injection. Compared with injection of either construct, the antibody titers were higher in the group with combined injection of two constructs.

OBJECTIVETo explore the effects of bone marrow mesenchymal stem cells (MSCs) on in vitro expansion potential, the adherent molecules expression of cord blood (CB) CD34(+) cells.

METHODSMSCs were obtained from human bone marrow and their differentiation function and phenotype were identified. CB CD34(+) cells were expanded in culture systems with or without MSC layer. Hematopoietic progenitor cells and adhesion molecules expression were assessed by semisolid culture assay and flow cytometry.

RESULTSThy-1, SH2, SB10, CD44, CD13, CD49e and CD29 were highly expressed on MSCs with no expressions of CD34, CD45, HLA-DR, CD14 and CD31. The MSCs could differentiate into adipocytes and osteoblasts under specific induction conditions. After culturing on MSCs layer with supplement of cytokines for 8 days, the absolute numbers of nuclear cells, CD34(+), CD34(+)CD38(-), CD34(+)CD62L(+) cells and CFU-Cs were increased by 145.57 +/- 17.89, 37.47 +/- 13.78, 69.78 +/- 50.07, 10.74 +/- 5.89 and 20.73 +/- 5.54-folds, respectively, being significantly higher than that cultured with cytokines alone. The expression of ALCAM, VLA-alpha4, VLA-alpha5, VLA-beta1, HCAM, PECAM and LFA-1 on CD34(+) cells remained unaffected. The expressions of ICAM-1 and L-selectin were downregulated during expansion, while the absolute numbers of CD34(+)CD62L(+) and CD34(+)CD54(+) cells were increased.

CONCLUSIONSMSCs layer improves expansion of CB CD34(+) cells while inhibiting their differentiation and retaining their homing ability.

Antigens, CD34 ; Cell Adhesion Molecules ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Mesenchymal Stromal Cells

OBJECTIVETo evaluate the effect of Taohong Siwu Tang and its fractions on hotplate-induced pain in mice, acetic acid-induced writhing response, dysmenorrheal model and isolated uterine contraction in vitro in mice, and discuss material basis of effect sites.

METHODThe samples of fractions were prepared by macroporous adsorptive resins (TH-1-TH-15). In the whole animal experiment, the hotplate-induced pain mice model was established to observe the effect of the samples on pain threshold in mice; the acetic acid-induced writhing response mice model was built to observe the effect of the samples on the writhing response in mice; the mice dysmenorrheal model was established to observe the effect of the samples on the writhing response, and calcium ion (Ca2+) and nitric oxide (NO) levels in uterine tissue of mice. In the isolated uterus contraction experiment, the oxytocin-induced isolated uterus contraction mice model was established to observe the effect of the samples on the isolated uterus contraction index. HPLC-DAD method was adopted for the content determination of effect sites.

RESULTAccording to the evaluation of the integration geological effect, beside TH-2 and TH-4, other three active fractions (TH-9, TH-10 and TH-11) extracted from Taohong Siwu Tang are the main effect sites. Their chemical components were analyzed and identified as monoterpene glycosides, phthalides, organic acids, etc.

CONCLUSIONThe effect sites of Taohong Siwu Tang on dysmenorrhea are TH-9, TH-10 and TH-11, which are 30% - 50% active fractions obtained from water-soluble small-molecular fractions by gradient elution using ethanol through macroporous absorption resin. Compared with TH-10 and TH-11, TH-9 shows stronger effect, which may be related to the type and content of chemical components it contains.

Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; therapeutic use ; Dysmenorrhea ; drug therapy ; Female ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred ICR

OBJECTIVETo constitute the animal model of unilateral olfactory nerve transection and observe the expression level and distribution of odorant receptors.

METHODSThirty-two rats were divided into two groups: the olfactory nerve transection group (20) and the control group (12). The former group received the operation to transect the left olfactory nerve following the left olfactory bulb was exposed under microscope and the latter group did not give any disposal. At every stage of five days, two weeks, four weeks and six weeks after the operation, five rats from the nerve transection group and three from the control group were anaesthetized simultaneously, and olfactory epithelium were taken out after transcardial perfusion, then paraffin imbedding. Coronal sections were sliced for HE staining to observe the thickness changes of the olfactory epithelium, and for in situ hybridization (ISHs) to investigate the expression of olfactory receptor genes (Olr287, Olr226, Olr1493 and Olr1654) in the epithelium, also to evaluate the changes of the expression level and location of the selected receptors during the regeneration of olfactory epithelium.

RESULTSHE staining showed that 5 days after the operation cell quantity and thickness of the olfactory epithelium decreased obviously, which increased gradually 2 or 4 weeks after operation. After 6 weeks' recovery, the thickness of the epithelium could reach the control level. The pattern of cell staining by ISH showed a specific spatial distribution along the anteroposterior (AP) and dorsoventral (DV) axis. Evidence suggested that odorant receptors were distributed in continuous and multiple overlapping bands in the normal or nerve transected-recovered epithelium rather than in the conventionally accepted three or four zones. The data also demonstrated that the distribution of sensory neuron types, as identified and defined by odorant receptor expression, was restored to normal or nearly so by 6 weeks after operation. Likewise, the numbers of probe-labeled neurons in the nerve transected-recovered had an obvious decrease 5 days after olfactory nerve transection. Reactive cells (x(-) +/- s) of Olr1493 in the operated side was (53.9 +/- 19.9), compared with (419.0 +/- 21.2) in the unoperated side, there was statistic significance between them (t = 63.960, P < 0.01). Reactive cells increased gradually according to the regeneration of the epithelium, and were nearly equivalent to the normal side 6 weeks later without significant differentiation (t = 2.600, P > 0.05), according to the absolute positive cells in the operated and unoperated side of (417.8 +/- 32.4) and (445.3 +/- 10.0) respectively.

CONCLUSIONThe regeneration of the sensory neurons and receptors, both the number and the distribution, can recover to normal after olfactory nerve transection.

Animals ; Male ; Olfactory Mucosa ; metabolism ; Olfactory Nerve ; metabolism ; surgery ; Olfactory Nerve Injuries ; Olfactory Receptor Neurons ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Odorant ; genetics ; metabolism

Objective To explore the expression of human brain-derived neurotrophic factor (hBDNF) and green fluorescent protein (GFP) in hBDNF-GFP gene-transfected rat neural stem cells (NSCs) and the changes in the biological characteristics of the transfected cells. Methods NSCs were transfected with a lentiviral vector carrying hBDNF and GFP genes (hBDNF-GFP-NSCs) or GFP gene only (GFP-NSCs), with normal NSCs as the control. The expression levels of hBDNF mRNA and hBDNF protein in all the 3 groups were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot, respectively. Enzyme-linked immunosorbent assay (ELISA) was performed to detect hBDNF level in the cell culture medium before and after hBDNF-GFP gene transfection. Dorsal root ganglion (DRG) neurons and NSCs were cultured with the supernatants of the transfected NSCs and normal NSCs, and the growth status of the DRG neurons was observed and the proportion of NSCs differentiating into neurons determined. Results Compared with GFP-NSCs and normal NSCs, hBDNF-GFP-NSCs showed obvious hBDNF overexpression at both rnRNA and protein levels 7 days after the transfection, hBDNF content in the supematant of hBDNF-GFP-NSCs culture increased significantly with time and peaked 5 days after the transfecfion (P<0.05). Four days after culture in hBDNF-GFP-NSCs supernatant, the DRG neurons and adherent NSCs extended cells processes, and the ratio of the NSCs differentiating into neurons was higher in cells cultured in hBDNF-GFP-NSCs supematant than in those culture in GFP-NSCs and normal NSCs supematants. Conclusion Lentivitus can be used as the vector for hBDNF and GFP gene transfection into NSCs, and hBDNF-GFP gene-transfected NSCs maintain the basic characteristics of NSCs and are capable of stable expression and secretion of hBDNF and GFP.