[Clinical study of 19 cases of steroid-refractory gastrointestinal acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation with fecal microbiota transplantation].

Objective: To investigate the clinical efficacy of fecal microbiota transplantation (FMT) for treating steroid-refractory gastrointestinal acute graft-versus-host disease (GI-aGVHD) . Methods: This analysis included 29 patients with hematology who developed steroid-refractory GI-aGVHD after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Huaian Hospital Affiliated to Xuzhou Medical University from March 2017 to March 2022. Among them, 19 patients underwent FMT treatment (the FMT group) and 10 patients did not (the control group). The efficacy and safety of FMT were assessed, as well as the changes in intestinal microbiota abundance, lymphocyte subpopulation ratio, peripheral blood inflammatory cytokines, and GVHD biomarkers before and after FMT treatment. Results: ① Complete remission of clinical symptoms after FMT was achieved by 13 (68.4%) patients and 2 (20.0%) controls, with a statistically significant difference (P<0.05). Intestinal microbiota diversity increased and gradually recovered to normal levels after FMT and FMT-related infections did not occur. ②The proportion of CD3(+) and CD8(+) cells in the FMT group after treatment decreased compared with the control group, and the ratio of CD4(+), regulatory T cells (Treg), and CD4(+)/CD8(+) cells increased (all P< 0.05). The interleukin (IL) -6 concentration in the FMT group was lower than that in the control group [4.15 (1.91-5.71) ng/L vs 6.82 (2.40-8.91) ng/L, P=0.040], and the IL-10 concentration in the FMT group was higher than that in the control group [12.11 (5.69-20.36) ng/L vs 7.51 (4.10-9.58) ng/L, P=0.024]. Islet-derived protein 3α (REG3α) was significantly increased in patients with GI-aGVHD, and the REG3α level in the FMT group was lower than that in the control group after treatment [30.70 (10.50-105.00) µg/L vs 74.35 (33.50-139.50) µg/L, P=0.021]. Conclusion: FMT is a safe and effective method for the treatment of steroid-refractory GI-aGVHD by restoring intestinal microbiota diversity, regulating inflammatory cytokines, and upregulating Treg cells.

[Mendelian randomization and its application in periodontitis].

Mendelian randomization is a causal inference method using genetic variations as instrumental variables, which skillfully takes advantages of the distributive randomness and timing priority of genetic variation, effectively avoiding confounding biases and reverse causalities in traditional observational researches. It has become a research hotspot in recent years. As a complex inflammatory disease, periodontitis is associated with many factors, but the cognitions about these associations are mostly based on traditional observational studies, lacking strong evidences to infer the causality. In order to bring up new research ideas in the periodontal field, this article mainly reviewed Mendelian randomization and its research progress in periodontitis.

[Preoperative treatment of uterine fibroids with low-dose mifepristone: a multicenter, randomized, double-blind, placebo-controlled, parallel-group study].

Objective: To evaluate the clinical efficacy and safety of oral mifepristone (10 mg/day) versus placebo in the preoperative treatment of uterine fibroids. Methods: This study was a multi-center, randomized, double-blind, placebo, parallel controlled trial. A total of 132 patients with uterine fibroids were randomly divided into study group and control group, with 66 cases in each group. The patients in the study group orally took 1 tablet/day of mifepristone (dose of 10 mg/tablet), the patients in the control group orally took 1 tablet/day of placebo, and both groups were treated for 3 months. The primary efficacy evaluation indicators were the change rate of maximum fibroid volume; the secondary efficacy evaluation indicators included amenorrhea rate, improvement of subjective symptoms and anemia; the safety evaluation indicators included the analysis of adverse events and changes in laboratory biochemical indicators. Results: At the end of treatment, the maximum leiomyoma volume was reduced by 25.97% (95%CI: -34.79%--15.95%) in the study group and reduced by 1.51% (95%CI: -13.03%-11.54%) in the control group. The change rate of the maximum leiomyoma volume before and after treatment in the study group was significantly greater than that in the control group, and the difference in the change rate of the maximum leiomyoma volume between the two groups was -24.84% (95%CI: -36.56%--10.94%), which was much higher than the 10% superiority threshold goal set by this study within the 95%CI interval. At the end of treatment, the complete amenorrhea rate [84% (52/62)], dysmenorrhea elimination rate [98% (61/62)], and menstrual blood loss disappearance rate [87% (54/62)] in the study group were significantly higher than those in the control group (all P<0.05). At the end of treatment, the mean hemoglobin [(131±13) g/L], red blood cell count [(4.5±0.4)×1012/L] and hematocrit (0.39±0.03) in the study group were significantly increased compared with the baseline, and the differences had statistical significance (all P<0.05); after treatment, the differences in the above three indicators between the two groups had statistical significance (all P<0.01). The serum estradiol level in the study group was significantly lower than that in the control group at the end of treatment, and the difference was statistically significant (P<0.01). There were no significant differences in follicle-stimulating hormone and cortisol levels before and after treatment between the two groups (P>0.05). The overall incidences of any adverse event were not significantly different between the two groups (all P>0.05). Abdominal pain was the most common adverse event in the study group [9% (6/65)], but the incidence was not significantly increased compared with the control group [3% (2/64); P>0.05]. Conclusion: Compared with placebo, oral mifepristone 10 mg/day is significantly superior to placebo in reducing the size of uterine fibroids and improving anemia, without significant adverse reactions, and could be used as a drug treatment for patients with of uterine fibroids before surgery.

Relative efficiencies of alternative preference-based designs for randomised trials.

Recent work has shown that outcomes in clinical trials can be affected by which treatment the trial participants would select if they were allowed to do so, and if they do or do not actually receive that treatment. These influences are known as selection and preference effects, respectively. Unfortunately, they cannot be evaluated in conventional, parallel group trials because patient preferences remain unknown. However, several alternative designs have been proposed, to measure and take account of patient preferences. In this paper, we discuss three preference-based designs (the two-stage, fully randomised, and partially randomised designs). In conventional trials, only the treatment effect is estimable, while the preference-based designs have the potential to estimate some or all of the selection and preference effects. The relative efficiency of these designs is affected by several factors, including the proportion of participants who are undecided about treatments, or who are unable or unwilling to state a preference; the relative preference rate between the treatments being compared, among patients who do have a preference; and the ratio of patients randomised to each treatment. We also discuss the advantages and disadvantages of these designs under different scenarios.

Characterization of microbial community profiles associated with quality of Chinese strong-aromatic liquor through metagenomics.

AIMS: Microorganisms in fermentation pits (FPs) play key roles for Chinese-strong-aromatic-liquor (CSAL) production. However, the microbial community in the FPs is still poorly understood. Here, the aim of this study was to reveal the diversity and potential functions of microbiota in FPs. METHODS AND RESULTS: Sequencing-by-synthesis-based metagenomic sequencing and annotation results revealed that the microbiota of FPs was primarily composed of Firmicutes (54·6%), Euryarchaeota (15·3%), Bacteroidetes (10·1%), Gammaproteobacteria (5·8%), Opisthokonta (5·7%) and Unclassified_Bacteria (2·3%). And 133 genera were identified as the dominant genera of this fermentative food. Lactobacillus, Sedimentibacter, Syntrophomonas, Methanoculleus, Methanobacterium, Bacillus, Clostridium, Galactomyces, Candida, Pichia, Penicillium and Aspergillus were defined as active populations for biosynthesizing the characteristic volatile compounds of CSAL. The study also revealed that the microbial community structures changed significantly with different cellar ages and over different geographical regions. (i) The presence of Bacteroidetes was the most distinctive feature that characterized the different FPs ages. (ii) Distinct contents of Gammaproteobacteria and Euryarchaeota were observed at different positions in the FPs. (iii) Euryarchaeota markedly contributed to the generation of the character of the liquors with distinct geographical associations. CONCLUSIONS: This study demonstrated that the changes of microbial communities determined the different quality characteristics of CSAL. SIGNIFICANCE AND IMPACT OF THE STUDY: This research contributes to a deeper understanding of the FPs microbial composition and shows a new microbial resource for biotechnological applications.

Activation of PPARγ/P53 signaling is required for curcumin to induce hepatic stellate cell senescence.

Activation of quiescent hepatic stellate cells (HSCs) is the major event in hepatic fibrogenesis, along with enhancement of cell proliferation and overproduction of extracellular matrix. Although inhibition of cell proliferation and induction of apoptosis are potential strategies to block the activation of HSCs, a better understanding of the senescence of activated HSCs can provide a new therapeutic strategy for prevention and treatment of liver fibrosis. The antioxidant curcumin, a phytochemical from turmeric, has been shown to suppress HSC activation in vitro and in vivo. The current work was aimed to evaluate the effect of curcumin on senescence of activated HSCs and to elucidate the underlying mechanisms. In this study, curcumin promoted the expression of senescence marker Hmga1 in rat fibrotic liver. In addition, curcumin increased the number of senescence-associated ß-galactosidase-positive HSCs in vitro. At the same time, curcumin induced HSC senescence by elevating the expression of senescence markers P16, P21 and Hmga1, concomitant with reduced abundance of HSC activation markers α-smooth muscle actin and α1(I)-procollagen in cultured HSCs. Moreover, curcumin affected the cell cycle and telomerase activity. We further demonstrated that P53 pharmacological inhibitor pifithrin-α (PFT-α) or transfection with P53 siRNA abrogated the curcumin-induced HSC senescence in vitro. Meanwhile, curcumin disruption of P53 leading to increased senescence of activated HSCs was further verified in vivo. Further studies indicated that curcumin promoted the expression of P53 through a PPARγ activation-dependent mechanism. Moreover, promoting PPARγ transactivating activity by a PPARγ agonist 15d-PGJ2 markedly enhanced curcumin induction of senescence of activated HSCs. However, the PPARγ antagonist PD68235 eliminated curcumin induction of HSC senescence. Taken together, our results provided a novel insight into the mechanisms underlying curcumin inhibition of HSC activation through inducing senescence.

Characterization of agronomic and quality traits and HSW-G5 compositions from the progenies of common wheat (Triticum aestivum L.) with different protein content.

High molecular weight glutenin subunits (HMW-GS) play an essential role in wheat processing quality. In this study, we evaluated the genetic pattern with HMW-GS composition between generations and examined whether agronomic and quality traits were correlated with each other. A wheat (Triticum aestivum L.) cultivar with high protein content and 2 cultivars with low protein content were subjected to a reciprocal cross. Sixteen agronomic and 4 quality characteristics were investigated. A total of 216 seeds from each F2 generation were chosen randomly and analyzed for HMW-GS composition using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Agronomic and quality characteristics were not significantly different between reciprocal crosses, indicating no cytoplasmic effect on the characteristics studied. The separation ratio of 2 HMW-GS loci was 9:3:3:1, indicating no linkage between any 2 loci. The novel HMW-GS N was detected in cultivar R145, which did not follow the Mendelian segregation ratio. A Glu-A1a(1) band was not detected in 1 individual from Tian8901xR145. Average grain weight per spike was significantly correlated with quality characteristics and may be a suitable criterion for selecting high protein content in wheat breeding programs.

Topographical distribution pattern of cervical intraepithelial neoplasia across the cervix.

OBJECTIVE: The distribution of cervical intraepithelial neoplasia (CIN) lesions across the cervix was determined. METHODS: A total of 575 women whose pathological diagnosis after cervical conization was confirmed as CIN were studied; 146 had low-grade CIN and 429 had high-grade CIN. CIN lesion location on the cervix was recorded using 12-h clock face notation. RESULTS: In both groups, 12 o'clock was the most common and 2 o'clock the least common lesion location. The most severe lesions were most often located at 8 o'clock and 7 o'clock, in the low- and high-grade groups, respectively. The 2 o'clock site was the least frequent site for the most severe lesion in both groups. Lesions were found more frequently on the posterior lip of the cervix than on the anterior lip, and on the right side of the cervix than on the left side, in both groups. CONCLUSIONS: The distribution of CIN lesions is not randomly distributed across the cervix. The 12, 8 and 7 o'clock sites, and the posterior lip and right side of the cervix, should be targeted during colposcopy-directed biopsy of patients with CIN lesions as this may improve diagnostic accuracy.

Ethanol-induced neurodegeneration in NRSF/REST neuronal conditional knockout mice.

The transcription regulator, neuron-restrictive silencer factor (NRSF), also known as repressor element-1 silencing transcription factor (REST), plays an important role in neurogenesis and various neuronal diseases such as ischaemia, epilepsy, and Huntington's disease. In these disease processes, neuronal loss is associated with abnormal expression and/or localization of NRSF. Previous studies have demonstrated that NRSF regulates the effect of ethanol on neuronal cells in vitro, however, the role of NRSF in ethanol-induced neuronal cell death remains unclear. We generated nrsf conditional knockout mice using the Cre-loxP system to disrupt neuronal expression of nrsf and its truncated forms. At postnatal day 6, ethanol significantly increased the expression of REST4, a neuron-specific truncated form of NRSF, in the brains of wild type mice, and this effect was diminished in nrsf conditional knockout mice. The apoptotic effect of ethanol was pronounced in multiple brain regions of nrsf conditional mutant mice. These results indicate that NRSF, specifically REST4, may protect the developing brain from ethanol, and provide new evidence that NRSF can be a therapeutic target in foetal alcohol syndrome (FAS).

Chiral auxiliaries for asymmetric radical cyclization reactions: application to the enantioselective synthesis of (+)-triptocallol.

[figure: see text] A series of epimeric 8-aryl menthyl derivatives 5a-d and 6a-l, prepared from the same chiral source (R)-pulegone, were employed as chiral auxiliaries in the asymmetric radical cyclization reactions of beta-keto esters mediated by Mn(OAc)3. Chiral precursors 8c and 8d provided the cyclization products 10c and 10d, respectively, as single isomers (dr > 99:1), whereas the cyclization of precursor 9k gave 13k with good stereoselectivity (dr = 24:1). Diastereomer 13e was employed as the key intermediate in the enantioselective synthesis of (+)-triptocallol in 90% ee.

[Analysis of pregnancy rates after treatment of tubal pregnancy with systemic methotrexate].

OBJECTIVE: To analyse the pregnancy rate after treatment of tubal pregnancy with the systemic methotrexate (MTX) injection. METHODS: From March 1985 to August 1999, 129 women with confirmed unruptured tubal pregnancy, and desiring to conceive were selected. Among them 60 women were successfully treated with systemic MTX, and 69 with unilateral salpingectomy. All cases were followed up for 1-15 years. RESULTS: The rates of subsequent intrauterine pregnancies (IUP) in the MTX group was 73% (44 cases) and of recurrent extrauterine pregnancies (EP) was 8% (5 cases). Among 69 patients treated by salpingectomy, the rates of IUP and EP was 70% (48 cases) and 4% (3 cases) respectively. The differences between the two groups were not statistically significant. CONCLUSIONS: The effect of conservative management with MTX was similar to those of salpingectomy. The rate of subsequent pregnancy did not increased.

[Application of high frequency radiosurgical knife in the treatment of cervical diseases].

OBJECTIVE: To investigate the indication, resection scope of loop electrosurgical excision procedure (LEEP) and pathological characteristics. METHODS: 176 cases with abnormal cytological and colposcopical findings, including cervical intraepithelial neoplasia (CIN) and atypical squamous cells of undermined significance (ASCUS), were studied. Conization of cervix was performed in 63 cases of > or = CIN2, of which 33 cases with LEEP and 30 by traditional electrosurgical knife (TEK) as controls. Cervical biopsy was performed in 113 cases of CIN1 and ASCUS, of which 60 cases by LEEP and 53 by cervical biopsy forceps (CBF) as controls. RESULTS: Three months after conization of cervix the cytological persistent rate of disease was 6.1% in LEEP group and 6.7% in TEK group (P > 0.05). The operation duration of LEEP is (5.0 +/- 0.5) min, and is much shorter than TEK [(15.0 +/- 0.4) min]. The bleeding and recovery time of the cervix are also shorter (P < 0.01). Three months after cervical biopsy the cytological persistent rate of diseases was only 1.7% in LEEP group, significantly less than that in CBF group (35.8%). CONCLUSION: LEEP is indicated to conization of cervix of CIN2-3 and cervical biopsy of CIN1 and ASCUS. It is safe and time-saving. The optimal depth of cervical conization and biopsy was 7 mm and 4 mm respectively in the cervix, and 15 mm and 4 mm respectively in the cervical tube. It can offer intact sample for pathological diagnosis, and has special value for detecting cervical microinvasive carcinoma and local minor carcinoma.

[A pathological study on the correlation of HPV infection with precancerous cervical lesions].

OBJECTIVE: To study the correlation of cervical condyloma and cancerous lesions with HPV infection. METHODS: Cervical biopsies and histopathological examinations were performed on 179 cases which had abnormal cervical cytological smears. PCR was used to study the HPV-DNA of 128 cases and in situ hybridization (ISH) was used to study 10 cases. RESULTS: 1. Morphologic observations. Most cervical condyloma cases were of the morphologically flat type (97.7%). Two koilocyte types were observed, the classical, or so called diagnostic koilocyte type (39.7%) and the atypical type (60.3%). Cervical condyloma often occurred together with cervical intraepithelial neoplasia (CIN, 42.5%). 2. PCR HPV DNA subtype analysis. Of the 58 cervical flat condyloma, 5 were PHV6/11 positive (8.6%) and 28 were HPV16/18 positive (48.3%). 86.1% of those with atypical koilocyte and 9.1% of those with diagnostic koilocyte had HPV16/18 infection. 66.7% of the lesions in which condyloma coexisted with CIN(2-3) had HPV16/18 infection. CONCLUSIONS: Most cervical condyloma lesions were of the flat type. The appearance of atypical koilocyte is correlated to HPV16/18 infection, which in turn is correlated to the degree of CIN malignancy.

[Relationship between histopathologic observation of cervical condyloma and human papillomavirus infection].

OBJECTIVE: To study the relationship between histopathologic changes of cervical condyloma and different subtypes human papillomavirus (HPV) infection. METHODS: 158 women with abnormal Pap smears diagnosed by computer assisted cytologic technique (CCT) including 71 cases with atypical squamous cells of undetermined significance (ASCUS), 65 cases with low-grade squamous intraepithelial lesions (LSIL), 17 cases with high-grade squamous intraepithelial lesions (HSIL) and 5 cases with squamous cancers, underwent directed biopsies under colposcopy and were simultaneously detected for HPV6/11, HPV16/18 DNA by polymerase chain reaction (PCR). Koilocytotisis in 73 cases with pathologically proven cervical condyloma were grouped into type I and II according to its atypical degree of nuclei. RESULTS: HPV16/18 infection rate among condylomas cases was 86.0%, which was significantly higher than that of type I (16.7%) (P < 0.01). In LSIL with type II koilocytotisis, HPV16/18 infection rate and abnormal mitotic figures (AMFs) occurrence were 85.7%, significantly higher than those in type I koilocytotisis or cervical intraepithelial neoplasia I. CONCLUSIONS: Type II koilocytotisis was correlated with HPV16/18 infection. LSIL with type II koilocytotisises, distinct atypical nuclei, also associated with high HPV16/18 rate and AMFs, therefore treatment and follow-up should be more aggressive.

[Detection and sequence analysis of the p53 gene mutation in epithelial ovarian cancer].

OBJECTIVE: To find the characteristics of p53 gene mutation in epithelial ovarian cancer and to analyze the relationship between p53 mutation and FIGO stage. METHODS: p53 mutations in exon 5 to 7 were detected by single-strand conformational polymorphism (SSCP) and sequencing technique. RESULTS: 8 of 46 tumor tissues demonstrated a SSCP band shift in the region of the gene. All of them have been characterized to represent DNA alterations by sequencing, including 8 point mutations (6 missence, 1 silent mutation and 1 in intron) and a 1-base pair insertion (introducing a stop codon downstream). Overall, 88.9% of mutation were transitions, and most of them are G-->A transitions (7/8, 87.5%). 62.5% of the mutation were found in 175 and 245 codon. The percentage of the mutation in stage I and stage II was 20.0%, and in stage III and stage IV was 16.7% (P > 0.05). CONCLUSION: The arising of p53 mutations in ovarian cancer is due to spontaneous error in DNA synthesis and repair. Codon 175, 245 are the two mutational hot spots. There is no relationship between the mutation of p53 gene and FIGO stage in epithelial ovarian cancer.

Amplifications of proto-oncogenes in ovarian carcinoma.

Thirty-two cases of ovarian carcinoma, two of normal ovaries, four of benign epithelial ovarian tumor, and three of borderline epithelial ovarian tumor were studied using Southern blot hybridization of DNA. In 15 of the 32 cases of ovarian carcinoma, peripheral lymphocytes were also studied. The amplification rate of C-myc, C-N-ras, C-Ki-ras and C-erbB-2 in ovarian carcinoma were 50%, 44%, 31% and 25% respectively. The amplification of C-Ki-ras and C-N-ras took place chiefly in cases of early stage and those of good differentiation. The amplification of C-N-ras was also found in cases of advanced stage. The amplifications of C-myc and C-erbB-2 were chiefly found in cases above stage III and those of poor differentiation. A total of 83% of the patients who died were found to have amplifications of more than 2 proto-oncogenes, with which the amplification of C-erbB-2 was involved.

[Amplification of proto-oncogenes C-myc, C-N-ras, C-Ki-ras, C-erbB2 in ovarian carcinoma].

OBJECTIVE: To observe the amplification rate of C-myc, C-N-ras, C-Ki-ras, C-erbB2 in human ovarian carcinoma. METHODS: Southern blot hybridization of DNA was employed, pathological diagnosis was made from fresh tissues. The beta-globin gene was used as an internal control. RESULTS: The amplification rate of C-myc, C-N-ras, C-Ki-ras and C-erbB2 in ovarian carcinoma was 50%, 44%, 31% and 25% respectively. The amplification rates of C-myc, C-erbB2 in stage III and IV were all significantly greater than that in stage I (P < 0.01). The amplification rate of C-N-ras in stage I was also significantly greater than that in stage III (P < 0.01). The amplification rate of C-Ki-ras in stage I was significantly greater than that in stage III or IV (P < 0.01). CONCLUSIONS: The amplification of C-Ki-ras and C-N-ras took place chiefly in cases of the early stages and in cases with good differentiation. The amplification of C-N-ras was also found in cases of advanced stages. The amplifications of C-myc and C-erbB2 were chiefly found in cases above stage III and in cases with poor differentiation, 83% of the patients who died were found to have amplifications of more than 2 proto-oncogenes, with the amplification of C-erbB2 involved in all of them.

Comparison of four methods to generate immunoreactive fragments of a murine monoclonal antibody OC859 against human ovarian epithelial cancer antigen.

In the present study, four different proteases (pepsin, papain, bromelain and ficin) were screened with a murine monoclonal antibody OC859, in order to verify whether different digestion procedures could improve yield and stability of the F(ab')2 or Fab fragments. The yields of F(ab')2 or Fab fragments from digestion with pepsin, papain, bromelain and ficin were respectively 20.3 +/- 2.0%, 50.5 +/- 5.0%, 74.4 +/- 2.7% and 82.8 +/- 10.2% of the theoretical maximum. Immunoreactivity in a noncompetitive solid-phase radioimmunoassay (SPRIA) of the fragments generated by the four proteases were respectively 10 +/- 5%, 36 +/- 5%, 60 +/- 6% and 75 +/- 6% of the intact OC859 IgG. These results suggested that the fragmentation of OC859 with ficin gave a higher yield of superior immunoreactive fragments.

[Serum lipid-associated sialic acid (LSA) in diagnosing and monitoring ovarian cancer].

Serum from 161 patients with ovarian cancer, 28 patients with benign gynecologic disorders and 22 healthy women, was assayed for levels of tumor marker LSA, which were compared with CA125. The results showed that in the patients with ovarian cancer, the sensitivities of LSA and CA125 for the patients prior to surgery were 83.0% and 92.5%, respectively; the sensitivities for the recurrent patients after surgery were 73.7% and 82.5% respectively. A total sensitivity of 89.5% was obtained by combination of both markers. The positive predictive value of LSA and CA125 for the patients with suspected tumor recurrence were 89.4% and 100%, respectively, and their corroborative rate with the postoperative courses were 94.4% and 100%, respectively. Thus serum assay of LSA, can be used in monitoring patients with ovarian cancer. The technique for determination of serum level of LSA is much more simple and less expensive than the radioimmuno-assay of serum level of CA125.