[Pathological diagnosis and clinical significance of appendiceal epithelial neoplasms].

Appendiceal epithelial neoplasms including benign and malignant, are clinically rare. There were quite a lot of changings in classification systems for them and different pathological diagnostic terminologies were used, resulted in confusions of understanding and communication for both pathologists and clinicians. Basically, appendiceal epithelial neoplasms include adenoma, serrated lesion and polyps, mucinous neoplasms, carcinoma and neuroendocrine neoplasms. Appendiceal mucinous neoplasms and goblet cell carcinoma are exclusively seen in the appendix. Though some appendiceal neoplasms are similar to those in large bowl, however, the molecular mechanism is different. The classification, pathological diagnosis and clinical significance of appendiceal epithelial neoplasms were summarized based on the fifth edition of WHO classification on digestive system tumors and other related literatures.

[Performance of combined liquid based cytology and HPV nucleic acid test for detecting cervical precancer among women attending screening].

Objective: To evaluate the clinical performance of HPV genotyping with cytology for detecting cervical precancer among women attending co-testing. Methods: A total of 2 883 females who participated in cervical cancer screening program were recruited from Erdos in 2016. All the participants were tested by cytology and HPV genotyping. In 2017, women with abnormal cytology results or HPV positive were followed up. Pathological cervical intraepithelial neoplasia (CIN) 2+ was the study end-point. Clinical performance indexes were calculated, including sensitivity, specificity, positive predictive value, negative predictive value, referral rate and missed cases. Results: INNO-LiPA resulted in a detection rate of 18.87%(544/2 883) for the 14-type high risk HPV. HPV16 was the most common infectious genotype (4.06%), followed by HPV52 (3.61%), HPV51 (2.50%), HPV58 (1.98%), and HPV18 (1.56%). With more HPV genotypes added into the group, sensitivity increased and the specificity decreased. Addition of HPV16, 58, 33, 39, 52, 18, 31 for detection lead to the maximun value of area under the curve (AUC)=0.913 (95%CI: 0.882-0.944). Compared with traditional screening method by cytology, cotesting decreased the number of missed diagnosis. Meanwhile, the fifth method (co-testing: triage of women with HPV16/18+ , cytological minor abnormalities and HPV58, 33, 39, 52, 31+ or cytological high grade abnormalities) did not increase referral rate (8.99% vs. 8.71%, P=0.525), with five cases of missed diagnosis (sensitivity of 92.1% and specificity of 93.2%). Conclusions: Co-testing with triage of women with HPV16/18+ , cytological minor abnormalities and HPV58, 33, 39, 52, 31+ or cytological high grade abnormalities would provide better clinical performance. In co-testing, triage of HPV16/18 was used in women with normal cytology; triage of HPV58, 33, 39, 52 and 31 was used in women with low-grade abnormal cytology; referral colposcopy was used in women with high-grade abnormal cytology, which would provide better clinical performance.

[Association between high risk human papillomavirus DNA load and cervical lesions in different infection status].

Objective: To explore the relationship between high risk HPV (HR-HPV) DNA load and cervical lesions in HR-HPV single/ multiple infections. Methods: Two thousand six hundred and forty-six women from Shanxi, Henan and Xinjiang were recruited into a cervical cancer screening program. Cervical exfoliated cell specimens collected from all of the participants were detected by hybrid capture Ⅱ (HC2), cytological diagnosis was performed according to the Bethesda System, and pathological diagnosis was interpreted using cervical intraepithelial neoplasia (CIN) terminology.Totally 571 cervical specimens were selected and retested to ascertain the HPV types and single/ multiple infections by liner array, a PCR-based method. Semi-quantitative result of HR-HPV DNA load (pg/ml) was estimated by HR HC2.According to the taxonomy of "International Human Papillomavirus Reference Center" , 13 HR-HPVs, including HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68, which could be detected by HR HC2 were divided into 4 subgroups. Results: The positive rate of HR-HPV in normal cervix (436 cases), CIN1 (88 cases), CIN2+ (47 cases) group were 29.82%, 85.23% and 100%, respectively. The overall prevalence and median viral load increased coordinating with the pathological degree of cervical lesions (P<0.001). The positive rate and viral load of single infection with HR-HPV belongs to α9 species increased coordinating with the pathological degree of cervical lesions (P<0.05). The viral load of single infection with HR-HPV belongs to α7 species in CIN1 group was higher than those of normal group and CIN2+ group, but without statistical significance (P=0.130). The viral load of multiple infections in CIN1 group was 559.13 pg/ml, significantly higher than 37.73 pg/ml of normal histology (P=0.025), but without significant difference of 332.91 pg/ml of CIN2+ group (P=0.790). The median viral load of HPV single infection in CIN1 group was 167.93 pg/ml, significantly lower than 559.73 pg/ml of multiple infections (P=0.044). The incidence of co-infection with HR-HPVs belong to α9 species was 80.56%, dominated in all patterns of multiple infections and their median viral load increased coordinating with the pathological degree of cervical lesions, but without significant difference (P>0.05). The incidence of co-infection with HR-HPVs belong to α7 species was 66.67%, their median viral load in CIN1 group was higher than that of CIN2+ group, but without statistical significance (P>0.05). Conclusions: Viral loads of single/ multiple infections with HR-HPVs belong to different species show different tendencies coordinating with the pathological degree of cervical lesions. Women with high grade of cervical lesion were dominantly infected with high viral load of HR-HPVs belong to α9 species, and the viral load of multiple infections is higher than that of single infection in low grade of cervical lesion.

Sub-millisecond (125)Te NMR spin-lattice relaxation times and large Knight shifts in complex tellurides: Validation of a quadratic relation across the spectrum.

(125)Te NMR spectra and spin-lattice relaxation times, T1, have been measured for several GeTe-based materials with Te excess. The spectra show inhomogeneous broadening by several thousand ppm and a systematic variation in T1 relaxation time with resonance frequency. The quadratic dependence of the spin-lattice relaxation rate, 1/T1, on the Knight shift in the Korringa relation is found to be valid over a wide range of Knight shifts. This result confirms that T1 relaxation in GeTe-based materials is mostly dominated by hyperfine interaction between nuclei and free charge carriers. In GeTe with 2.5% excess of Te, about 15% of the material exhibits a Knight shift of ≥4500ppm and a T1 of only 0.3ms, indicating a high hole concentration that could correspond to close to 50% vacancies on the Ge sublattice in this component. Our findings provide a basis for determining the charge carrier concentration and its distribution in complex thermoelectric and phase-change tellurides, which should lead to a better understanding of electronic and thermal transport properties as well as chemical bonding in these materials.

[Distribution of human papillomavirus (HPV) among HPV positive cervical adenocarcinoma cases detected by laser capture microdissection(LCM)].

OBJECTIVE: To investigate the distribution of human papillomavirus (HPV) in the diseased areas cut from HPV-positive cervical adenocarcinoma (ADC) detected by laser capture microdissection (LCM). METHODS: Paraffin-embedded specimens diagnosed as ADC between 2005 and 2010 were collected from 9 hospitals in 7 regions across China. HPV genotyping was conducted on paraffin sections using sandwich technique and LCM in order to identify HPV infection in the tumor tissues. HE and p16 immunohistochemistry staining were performed to make histological diagnosis. RESULTS: A total of 169 cervical adenocarcinoma cases were recruited, including 94 cases of mucinous adenocarcinoma (ADC-CX), 9 cases of adenosquamous carcinoma (ASC), 19 cases of minimal deviation adenocarcinoma (ADC-MIN), 14 cases of clear cell adenocarcinoma (ADC-CC), 8 cases of endometrioid adenocarcinoma (ADC-ENDO), 9 cases of serous adenocarcinoma (ADC-SER) and 16 cases of adenocarcinoma not otherwise specified (ADC-NOS). Fourteen types of high risk HPV were detected in the whole tissue section (WTS). HPV16 was the most common type, and the second was HPV18 and HPV52, respectively. Compared with WTS, the HPV-positive rate detected by LCM was lower. The HPV positive rates were significantly different among different subtypes of cervical adenocarcinoma (P<0.001). After LCM, the HPV positive rate was 50.8% and 66.7% in the single infection and multiple, infection groups respectively (P=0.14). The positive rates of p16 was significantly different among different subtypes of cervical adenocarcinoma (P<0.001). p16-positive rate was 73.9% in the HPV-positive samples after LCM, significantly higher than the 38.5% of negative samples (P<0.001). CONCLUSIONS: Laser capture dissection technique can more precisely reflect the HPV distribution in cervical adenocarcinomas. The etiological association between HPV infection and cervical adenocarcinoma occurrence is not as close as that reported in the literature.

Diagnostic potential of serum protein pattern in Type 2 diabetic nephropathy.

AIMS: Microalbuminuria is the earliest clinical sign of diabetic nephropathy (DN). However, the multifactorial nature of DN supports the application of combined markers as a diagnostic tool. Thus, another screening approach, such as protein profiling, is required for accurate diagnosis. Surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) is a novel method for biomarker discovery. We aimed to use SELDI and bioinformatics to define and validate a DN-specific protein pattern in serum. METHODS: SELDI was used to obtain protein or polypeptide patterns from serum samples of 65 patients with DN and 65 non-DN subjects. From signatures of protein/polypeptide mass, a decision tree model was established for diagnosing the presence of DN. We estimated the proportion of correct classifications from the model by applying it to a masked group of 22 patients with DN and 28 non-DN subjects. The weak cationic exchange (CM10) ProteinChip arrays were performed on a ProteinChip PBS IIC reader. RESULTS: The intensities of 22 detected peaks appeared up-regulated, whereas 24 peaks were down-regulated more than twofold (P < 0.01) in the DN group compared with the non-DN groups. The algorithm identified a diagnostic DN pattern of six protein/polypeptide masses. On masked assessment, prediction models based on these protein/polypeptides achieved a sensitivity of 90.9% and specificity of 89.3%. CONCLUSION: These observations suggest that DN patients have a unique cluster of molecular components in serum, which are present in their SELDI profile. Identification and characterization of these molecular components will help in the understanding of the pathogenesis of DN. The serum protein signature, combined with a tree analysis pattern, may provide a novel clinical diagnostic approach for DN.

Gas chromatographic-mass spectrometric determination of 20(S)-protopanaxadiol and 20(S)-protopanaxatriol for study on human urinary excretion of ginsenosides after ingestion of ginseng preparations.

An improved gas chromatographic-mass spectrometric method (GC-MS) with a fast solid-phase extraction on a newly introduced C18 microcolumn, was applied to study the urinary excretion 20(S)-protopanaxadiol and 20(S)-protopanaxatriol glycosides in man after oral administration of ginseng preparations. Using panaxatriol as internal standard, 20(S)-protopanaxadiol and 20(S)-protopanaxatriol (the aglycones of ginsenosides) could be determined at a detection level of a few ng per ml urine by GC-MS with selected-ion monitoring after their release from glycosides which occur in urine. The extraction recovery of ginsenosides from urine was more than 80% and the intra-assay coefficient of variation was less than 5.0%. The results after intake of single doses of ginseng preparations demonstrated a linear relation between the amounts of ginsenosides consumed and the 20(S)-protopanaxatriol glycosides excreted in urine. About 1.2% of the dose was recovered in five days.

Determination of aglycones of ginsenosides in ginseng preparations sold in Sweden and in urine samples from Swedish athletes consuming ginseng.

Recently developed gas chromatographic and gas chromatographic-mass spectrometric methods were used to characterize 17 different commercial ginseng preparations sold in Sweden. The contents of total ginsenosides per capsule or per tablet varied from 2.1 to 13.3 mg. Unlike the other preparations, a red ginseng and three liquid ginseng preparations (after releasing the sugar moieties from ginsenosides) were shown also to contain significant amounts of 20-epimers of 20(S)-protopanaxadiol and 20(S)-protopanaxatriol as well as their corresponding 24,25-hydrated compounds. In addition to the genuine and artificial sapogenins mentioned above, two epimeric pairs of prosapogenines (ginsenoside Rg3 and 20(S)-Rg3, ginsenoside Rh1 and 20(R)-Rh1) were also found in the liquid formulations. These results suggest that hydrolysis, epimerization and hydration in the side-chain of the aglycone moiety of ginsenosides may occur in the liquid formulations under weak acidic conditions (pH 3.0-3.5 with 9-10% of alcohol at room temperature). The new method was also used to determine the aglycones of ginsenosides in urine samples from Swedish athletes stating that they had consumed ginseng preparations within 10 days before urine collection. Out of the 65 samples analysed, 60 were found to contain 20(S)-protopanaxatriol. The concentrations of 20(S)-protopanaxatriol ginsenosides varied from 2 to 35 ng ml-1 urine. This is the first demonstration of uptake of ginsenosides in humans after oral administration of ginseng preparations.

Analysis of ginsenosides by chromatography and mass spectrometry: release of 20 S-protopanaxadiol and 20 S-protopanaxatriol for quantitation.

To facilitate studies on the possible presence of ginseng products in serum, tissues, and excretions, a procedure to optimize the analysis of the ginseng specific products, i.e., ginsenosides, had to be worked out. With the present method the two sapogenins, 20S-protopanaxadiol and 20S-protopanaxatriol, can be produced from ginsenosides Rb1, Rc, Rd, Re, and Rg1 in 80% yield by using an improved alkaline cleavage procedure. In contrast to previously described acid hydrolysis procedures for ginsenosides, our alkaline conditions caused no epimerization, no hydroxylation, and no cyclization of the side chain. Furthermore, no unchanged ginsenosides were recovered. The products of alkaline and acidic cleavage were separated, identified, and characterized by GC, GC-MS, and HPLC. In contrast to alkaline cleavage, treatment with acid afforded a number of side products. The C-20S-epimers of the ginseng sapogenins could be distinguished from C-20R epimers by difference in mass spectra and retention time after trimethylsilylation.

[Separation and identification of stimulants and their metabolites].

Forty-one stimulant drugs banned by the International Olympic Committee were studied after they were administered to human volunteers. The parent drugs and their metabolites in free or conjugated forms in human urine collected within 24 hours after administration of the drugs were extracted, separated and identified. The separation was performed on a capillary gas chromatograph with nitrogen-phosphorous detector, while the identification was achieved on a capillary gas chromatograph with mass-selective detector. The extract was injected into the gas chromatograph both directly and after derivatization with trifluoroacetic anhydride (TFAA) or N-methyl-N-trimethylsilyl trifluoroacetamide (MSTFA) as well as TFAA and MSTFA combined. The conjugated metabolites were studied after acid hydrolysis of the extract and then selectively derivatized as above. The results are summarized in 4 tables in the text.

[The identification of ephedrine and its analogues in urine by gas chromatography].

A sensitive and reliable method of analysis for ephedrine and its analogues was established by using GC and GC-MS. The columns used for both GC and GC-MS were HP-5, and the carrier gas was helium. NPD was used as the detector for GC. The urine sample was extracted with ether and submitted to GC. If positive, it was then submitted to GC-MS to identify its structure. The detection limits for ephedrine and its analogues are lower than 200 ng/ml urine. The recoveries are above 80%.

[Determination of six Ephedra alkaloids in Chinese Ephedra (Ma Huang) by gas chromatography].

Six Ephedra alkaloids, namely ephedrine, pseudoephedrine, norephedrine, norpseudoephedrine, methylephedrine and methylpseudoephedrine, in 12 species of Chinese Ephedra were successfully separated and determined by gas chromatography with the highly specific and sensitive nitrogen phosphorus detector (GC/NPD). The column used (HP-5) had a cross linked 5% phenylmethylsilicone phase. Diphenylamine was used as the internal standard to check the reproducibility of the extraction yields of the alkaloids, the stability of the detector response and to quantify the alkaloids. The contents of the six alkaloids were calculated according to their regression equations. The way for the preparation of crude drug samples was improved, the diethyl ether extract of the alkalized crude sample was directly analysed by GC. The method is simple, rapid and sensitive. The results are in agreement with those of the HPLC method.

[Study on metabolism of phenmetrazine-like drugs and their metabolites].

We have studied the metabolism of phenmetrazine, phendimetrazine and morazone, and their metabolites in urine. A sensitive, specific method using GC/NPD was applied to the determination of the rate of excretion of the drugs quantitatively, diphenylamina being used as the internal standard. The ether extract was derivatized with TFAA reagent, the metabolites and their TFA derivatives were identified by GC/NPD and GC/MSD methods. Norephedrine, cathine, ephedrine and pseudoephedrine in small amount were identified.

[Studies on the medicinal plants of Magnoliaceae tu-hou-po of Manglietia].

Data from a survey of the drug market and investigation of the original plant of "Tu-hou-po", after careful botanical examinations, showed that the drugs were derived from 5 species of the genus Manglietia of Magnoliaceae, viz. Manglietia chingii Dandy, M. insignis (Wall.) Bl., M. duclouxii Finet et Gagnep., M. yuyuanensis Law and M. szechuanica Hu. Comparisons of the main characteristics of the plants, Tu-hou-po and Hou-po crude drugs and chemical components showed that Manglietia is taxonomically the closest to Magnolia and contained similar components (tab 1-2 and fig 1). The results of HPLC analysis demonstrated that they contained magnolol, honokiol, magnocurine and salicifoline, in different quantities. However, no magnosprengerine was detected. Besides, it was also found that the percentage of magnolol and honokiol contents were higher, while that of magnocurine was lower in Hou-po. On the contrary, the content of magnocurine was higher, while that of magnolol and honokiol were lower in Tu-hou-po. Manglietia chingii (Tu-hou-po) is being used as the Chinese traditional drug "Hou-po" in the clinic in Guangxi. Therefore, M. chingii is noteworthily exploited as a new resource of Hou-po for further research.