Objective To study changes of DNA content in human spleen nuclei and seek an experimental method for estimation of the postmortem interval(PMI)using computerized image-analysis technique(CIAT).Methods Smear sections from spleen sampled were collected in 36 cadavers with known the accurate PMI respectively every hour within the first 36 hours after death,which were then fixed with cold Carony fixation.The smeared sections were stained by Feulgen-van's staining method.3 indices for spleen nucleic DNA including integral optical density(IOD),average optical density(AOD)and average gray(AG)were measured using the CIAT.Results IOD and AOD in the spleen nuclei declined regularly,whereas AG increased with the extension of PMIs in 36 hours.Conclusion There are definite relationships between the PMI and gray parameters(IOD、AOD and AG)representing the DAN content of nucleic DNA in the spleen in 36 hours after death,which may be used for estimation of PMI.

Objective To search for the optimal processing procedure and physico-chemical properties of Endothelium corneum amylase. Methods After dried under 50 ℃ ,amylase was extracted from Endothelium corneum with phosphate-buffered saline. The amylase activity was examined at different temperature,pH and different metal ions,and amylase isozyme was discerned on PAGE. Results The optimal pH,optimal temperature,and optimal substrate concentration for amylase activity were 8.67,50 ℃ and 1.8 g/mL,respectively. Under the experimental conditions,the amylase Km was 0.92 (starch was used as substrate of amylase). Four metal ions tested,including Ca2+,Fe2+,Zn2+,Mn2+,could inhibit the amylase activity to some extent. Furthermore,it showed one amylase isozyme band using PAGE,suggesting only one amylase isozyme existing in the Endothelium corneum tentatively. Conclusion The effects of metal ions should be considered in the process of Endothelium corneum amylase.

Group A,the differences among the three groups were statistically significant(P0.05). TCR,CCR,DAP,OD,BRC in Groups B and C showed trends of increasing,the differences in terms of the contents of BMP-RⅠamong the 3 phases were statistically significant(P0.05);the ER in Group B and C showed a trend of decreasing,thee difference between 4- and 8-week and 4- and 12-week were signifieantly dif- ferent(P0.05).Conclusion After application of glucocorticoid for a short term,pathological changes maintained and showed trends of inereasing in early stage.HBO can reverse these changes.The outcome of 3-course HBO therapy is better than that of 1-course therapy.

Objective To investigate whether cesarean section on pregnant women with chronic hepatitis B virus (HBV) infection may reduce mother-to-infant transmission risk of HBV.Methods Preserved serum samples of pregnant women at 15-20 weeks of gestation which were collected from July 2002 to August 2004 in 14 counties of Jiangsu Province were analyzed retrospectively.These samples were tested for HBV serologic markers with enzyme immunoassay,and 419 cases of positive hepatitis B surface antigen (HBsAg) were found.HBV DNA level of these HBsAg-positive women were further quantitatively measured with real-time polymerase chain reaction assay.Children of these 419 HBsAg-positive mothers were followed up during October 2009 to March 2010 and 298 of them were contactable (71.1％),among which 281 were enrolled in final analysis as they had complete information and had been vaccinated with three regular doses of HBV vaccine during infantile period.The 281 children were tested for serum HBV markers.The HBV infection rate in these children at age 5-7 was compared between those born by cesarean setion (CS group,n =136) and by vaginal delivery (VD group,n=145).T-test,x2 test or Fisher's exact test was used to analyze relevant parameters.Results(1) Maternal HBeAg-positive rates [25.7％ (35/136) vs 34.5％ (50/145)],HBV DNA levels in HBeAg-positive women [(2.30 × 106) IU/ml vs (2.09× 106) IU/ml],the percentages of HBIG injection after birth [38.2％ (52/136) vs 35.9％ (52/145)],breastfeeding ratios [82.4％ (112/136) vs 75.9％ (110/145)],and children' s ages at the follow-up [(5.9 ± 0.8) vs (6.0±0.6)] were comparable between CS group and VD group (all P＞0.05).(2) Among these 281 children,272 (96.8％) were HBsAg-negative,9 (3.2％) were HBsAg-positive and 163 (58.0％) were anti-HBs-positive.The prevalence of HBsAg-positive [(2.9％ (4/136) vs 3.4％ (5/145)],HBsAg negative and anti HBc positive [0.0％ (0/136) vs 1.4％ (2/145)] and anti-HBs positive [57.4％(78/136) vs 58.6％(85/145)] in CS group were similar to those in VD group.Conclusions Cesarean section does not reduce the risk of mother-to-infant transmission of HBV.In clinical practice,elective cesarean section should not be suggested in order to prevent HBV vertical infection.

AIM: To detect the serum proteomic patterns in patients of breast cancer by the method of SELDI-TOF-MS and CM10 ProteinChip, and to screen the biomarker candidates, build and validate the diagnostic models, and evaluate its clinical value in surveillance and follow-up after operation. METHODS: The SELDI-TOF-MS technology and CM10 ProteinChip were used to detect the proteomic patterns of serum from 63 breast cancer patients and 40 healthy women. The biomarker candidates were screened and the diagnostic models were constructed by ZJU-PDAS software. Meanwhile, the model was blind-validated in another 23 patients and 20 healthy women. At the same time, 16 serum samples were detected to evaluate its value in surveillance and follow-up after operation. RESULTS: The best model was composed by two protein peaks (BC1/3.9 kD and BC2/5.6 kD) with its sensitivity and specificity of 87.30% (55/63) and 95.00% (38/40), respectively. The sensitivity and specificity in the blind-validation of new cases were 95.65% (22/23) and 85.00% (17/20), respectively. The diagnostic efficacies were the same to the patients of different stages (P>0.05). The expression of BC1 increased while BC2 decreased after operation. The expression of BC2 in the patients with recurrence or metastasis was higher than that in the tumor-free survivors (P<0.05). CONCLUSION: This method shows its potential in detection, surveillance and follow-up after operation. The method is also useful for screening the novel and better biomarkers in breast cancer.

ObjectiveTo explore the relations between the alkaline phosphatase(ALP) and the type of cerebral palsy. MethodsThe ALP level of 40 children with different types of cerebral palsy were examined with the Olympus AU-600 apparatus. ResultsThe ALP value of the children with spastic cerebral palsy is normal. The ALP value of the children with athetoid cerebral palsy is higher than spastic cerebral palsy(P<0.001).Conclusions The ALP value can help us to identify the type of cerebral palsy.

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.

OBJECTIVETo reveal the quality variation of polysaccharides, triterpenoids and proteins in spores and fruiting bodies of Ganoderma lucidum from producing areas, different varieties, harvesting parts and periods, and wall-breaking treatments.

METHODSpores and fruiting bodies from varieties of Longzhi No. 1 and Hunong No. 1 were collected as test samples, together with wall-broken spores sold in domestic main producing areas. The anthrone-sulfuric acid colorimetric method was used to determine the content of total polysaccharides. The vanillin-glacial acetic acid-perchloric acid colorimetric method was used to determine the content of total triterpenoids. The Lowry method was used to determine the content of total proteins.

RESULTThe content ranges of total polysaccharides, total triterpenoids, and total proteins from 6 domestic main producing areas were 0.40% - 2.25%, 1.36%-3.15% and 0.74% -1.91% respectively. The content ranges of total polysaccharides, triterpenoids, and proteins in the fruiting bodies from 2 varieties cultured in Zhejiang were 0.25% -1.42%, 0.44% -1.42% and 1.82% -3.67% respectively. In addition, the ranges of samples from wall-unbroken spores were 0.41% - 0.91%, 0.09% - 0.12%, 0.78% - 0.90% respectively and wall-broken spores are 1.03% - 2.25%, 1.89% - 3.15%, 0.96% - 1.04% respectively.

CONCLUSIONThere are significant differences in the contents of main chemical ingredients of wall-broken G. lucidum spores saled in the markets. The samples from Zhejiang contain high content of total polysaccharides and triterpenoids, and samples from Fujian contains more proteins. Between the 2 major varieties cultured in Zhejiang, Longzhi No. 1 contains higher content of triterpenoids, but Hunong No. 1 has more polysaccharides. Contents of triterpenoids and polysaccharides from wall-broken spores are much higher than those of fruiting bodies. The stipes from fruiting bodies contains more polysaccharides than those of the pileus, while the triterpenoids contents are higher in the pileus than stipes. The pileus and stipes collected in the second year contain higher content of polysaccharides than the first year's samples, but the contents of triterpenoids are lower. Wall-breaking treatment would significantly improve the extraction and dissolution rate of total triterpenoids and polysaccharides.

Fungal Proteins ; analysis ; Polysaccharides ; analysis ; Reishi ; chemistry ; Spores, Fungal ; chemistry ; Triterpenes ; analysis

Objective To assess the protective effect of vaccination in routine application on hepatitis B virus (HBV) exposed infants and to clarify whether hepatitis B immunoglobulin (HBIG) administration of pregnant women may reduce the risk of maternal-fetal transmission of HBV. Methods Serum samples of 6398 pregnant women at gestation of 15-20 weeks from 6 urban and 8 rural areas across Jiangsu province were previously tested for serologic markers of HBV by ELISA from July 2002 to August 2004. In this study, infants born to 419 HBV carrier mothers were taken as the study group, while infants born to 453 non-carrier mothers were taken as the control group by stratified random sampling. They were followed-up and screened for HBV markers during October 2009 to March 2010. Information including HBIG administration during pregnancy, HBV vaccination and HBIG administration of the infants were collected. χ2 test or Fisher′s exact method were used to compare the rates and the comparison of the means was by t test. Results The follow-up rates of the study group and control group were 71.12% (298/419) and 72.41% (328/453), respectively. Of the 298 infants born to HBV carrier mothers, 11 (3.7%) were positive for HBsAg, while none of the 328 infants born to non-carrier mothers was HBsAg positive (χ2=12.32, P<0.01). All of the 11 children were born to mothers with both HBsAg and HBeAg positive, and nine of the 11 children were not injected HBIG or not immunized with hepatitis B vaccine within 24 hours after birth, with only one received regular vaccination and detailed information was unknown in one case. The positive rates of anti-HBs in the study group and the control group were 69.46% and 69.21% respectively (χ2=0.01, P=0.95). HBsAg positive rate of the children born to pregnant women treated with HBIG during late pregnancy (n=92) was 2.17% (n=2), whereas that in the children born to women not treated with HBIG (n=197) was 4.57% (χ2=0.98, P=0.51). Conclusions The protective effect of immunoprophylaxis in routine application against perinatal HBV infection in Jiangsu province is good. Efforts are required to emphasize the importance of HBIG administration in infants born to HBV carrier mothers, especially in HBeAg positive mothers within 24 hours after delivery. Treatment of HBsAg positive pregnant women with HBIG in third trimester would not decrease the risk of maternal-fetal transmission of HBV.

ObjectiveTo investigate the relationship between the expression of ER,PR,HER-2,Ki-67 and the effective rate of neoadjuvant chemotherapy (NAC) and to study the influence of NAC on expression of ER,PR,HER-2 and Ki-67 in breast cancer.Methods41 patients of breast cancer were performed 2 to 6 cycles of CAF chemotherapy.The expression of ER,PR,HER-2 and Ki-67 was detected by SP immunohistochemical method before and after NAC.ResultsThe effective rate of NAC was 73.17％ (30/41).The positive and negative efficiency rate was 67.86％ (19/28) vs 84.62％ ( 11/13 ) for ER,64.29％ (9/14) vs 77.78％ (21/27 ) for HER-2,70.00％ (14/20) vs 76.19％ ( 16/21 ) for Ki-67.The difference had no statistical significance (P ＞0.05).The positive and negative efficiency rate was 66.67％ (16/24) vs 82.35％ (14/17) for PR.The difference had statistical significance (P ＜ 0.05 ).The expression of ER,PR,HER-2 and Ki-67 before and after NAC changed in 10,8,3 and 9 cases respectively.The difference had no statistical significance (P ＞ 0.05).ConclusionsNAC has no influence on the expression of ER,PR,HER-2 and Ki-67.Patients with PR negative are more sensitive to NAC.The expression state of PR can be used to predict the curative effect of NAC.The expression state of ER,HER-2 and Ki-67 has no significant correlation with NAC.