[Linggui Formula for asthenospermia with kidney deficiency and blood stasis: A randomized controlled trial].

OBJECTIVE: To observe the effect and safety of the traditional Chinese medicine (TCM) Linggui Formula (LGF) in the treatment of asthenospermia with kidney deficiency and blood stasis. METHODS: This randomized controlled trial included 90 cases of asthenospermia with kidney deficiency and blood stasis treated in our hospital from September 2022 to September 2023, 45 by oral medication with LGF (the trial group) and the other 45 with oral levocarnitine solution (the control group), all for 12 weeks. We followed up the patients for 12 weeks, recorded the semen parameters, TCM syndrome scores, sexual hormone levels, pregnancy rates, and DNA fragmentation index (DFI) of the patients, and compared them between the two groups before and after treatment. RESULTS: Totally, 82 of the patients completed the study, 42 in the trial and 40 in the control group. After treatment, the patients in the trial group showed significant increases in the percentage of progressively motile sperm (PMS) (from ï¼»19.25±3.08ï¼½% to ï¼»38.57±4.99ï¼½%, P< 0.05), total sperm motility (from ï¼»32.29±3.64ï¼½% to ï¼»46.50±4.77ï¼½%, P< 0.05) and sperm concentration (from ï¼»83.9±37.2ï¼½ to ï¼»95.1±34.9ï¼½× 106/ml ï¼½, P< 0.05), and so did the controls in PMS (from ï¼»19.75±4.28ï¼½% to ï¼»34.46±5.07ï¼½%, P< 0.05), total sperm motility (from ï¼»33.02±4.93ï¼½% to ï¼»43.11±4.72ï¼½%, P< 0.05) and sperm concentration (from ï¼»85.2±39.7ï¼½ to ï¼»88.1±35.2ï¼½ × 106/ml , P< 0.05), all even more significant in the trial than in the control group (P< 0.05). No statistically significant difference was observed in the semen volume either in the trial (from ï¼»3.38±0.38ï¼½ to ï¼»3.24±0.45ï¼½ ml, P> 0.05) or in the control group (from ï¼»3.46±0.52ï¼½ to ï¼»3.30±0.37ï¼½ ml, P> 0.05), or between the trial and control groups (P> 0.05), or in the sexual hormone levels, pregnancy rates, and sperm DFI between the two groups before and after treatment (P> 0.05). Both groups of patients had good safety profiles without serious adverse reactions. CONCLUSION: Linggui Formula can improve the percentage of PMS in asthenospermia patients with kidney deficiency and blood stasis, potentially enhancing pregnancy rates and with a good safety.

Overexpression of Forkhead box Q1 correlates with poor prognosis in papillary thyroid carcinoma.

OBJECTIVE: Forkhead box Q1 (FOXQ1), a member of the forkhead transcription factor family, plays important parts in cell cycle, apoptosis, metabolism, immunology and tumour genesis. Its expression has been associated with poor clinical prognosis in various tumours. However, the clinical significance of FOXQ1 in papillary thyroid carcinoma (PTC) has not been fully studied. The purpose of this study was to investigate whether FOXQ1 is correlated with poor prognosis in PTC. DESIGN/METHODS: We performed a retrospective study of 136 PTCs. Immunohistochemistry (IHC) was used to examine the expression of FOXQ1 in 136 PTCs and 47 nodular goitre specimens. Rank-sum test, chi-square test, Kaplan-Meier survival analysis, univariate and multivariate Cox analyses were used to investigate the clinical and prognostic significance of FOXQ1 expression in PTC. RESULTS: The comparison of PTC specimens with nodular goitre with papillary hyperplasia specimens revealed an upregulation of FOXQ1 in PTC. Overexpression of FOXQ1 was observed in 63.24% of PTC and correlated with classic variant, tall variant, distant metastasis, AJCC stage and recurrence. FOXQ1-positive expression was associated with shorter disease-free survival: median disease-free survival of FOXQ1-positive patients was 23 months compared with 128 months for FOXQ1-negative patients (Log-rank χ2  = 12.31, P = 0.00045). Additional independent risk factors in this study were multifocality (recurrence-free survival [RFS]: hazard ratio [HR] = 2.391, P < 0.05), extrathyroidal extension (RFS: HR = 3.906, P < 0.05) and positive expression of FOXQ1 (RFS: HR = 6.385, P < 0.01). CONCLUSIONS: Our results indicated that FOXQ1 may be a useful additional biomarker to evaluate the progression of PTC and to predict likely relapse of disease.

Tanshinone IIA therapeutically reduces LPS-induced acute lung injury by inhibiting inflammation and apoptosis in mice.

AIM: To study the effects of tanshinone IIA (TIIA) on lipopolysaccharide (LPS)-induced acute lung injury in mice and the underlying mechanisms. METHODS: Mice were injected with LPS (10 mg/kg, i.p.), then treated with TIIA (10 mg/kg, i.p.). Seven hours after LPS injection, the lungs were collected for histological study. Protein, LDH, TNF-α and IL-1ß levels in bronchoalveolar lavage fluid (BALF) and myeloperoxidase (MPO) activity in lungs were measured. Cell apoptosis and Bcl-2, caspase-3, NF-κB and HIF-1α expression in lungs were assayed. RESULTS: LPS caused marked histological changes in lungs, accompanied by significantly increased lung W/D ratio, protein content and LDH level in BALF, and Evans blue leakage. LPS markedly increased neutrophil infiltration in lungs and inflammatory cytokines in BALF. Furthermore, LPS induced cell apoptosis in lungs, as evidenced by increased TUNEL-positive cells, decreased Bcl-2 content and increased cleaved caspase-3 content. Moreover, LPS significantly increased the expression of NF-κB and HIF-1α in lungs. Treatment of LPS-injected mice with TIIA significantly alleviated these pathological changes in lungs. CONCLUSION: TIIA alleviates LPS-induced acute lung injury in mice by suppressing inflammatory responses and apoptosis, which is mediated via inhibition of the NF-κB and HIF-1α pathways.

[Protective effect of formula of removing both phlegm and blood stasis on myocardial tissues of Chinese mini-swine with coronary heart disease of phlegm-stasis cementation syndrome].

OBJECTIVE: To study the protective effect of formula of removing both phlegm and blood stasis (TYTZ) on myocardial tissues of Chinese mini-swine with coronary heart disease of phlegm-stasis cementation syndrome. METHOD: Totally 36 Chinese mini-swine were randomly divided to six groups: the normal control group, the model group, the Danlou tablet group, and TYTZ groups with doses of 2.0, 1.0, 0.5 g x kg(-1), with six in each group. Except for the normal control group, all of other groups were fed with high-fat diet for 2 weeks. Interventional balloons are adopted to injure their left anterior descending artery endothelium. After the operation, they were fed with high-fat diet for 8 weeks to prepare the coronary heart disease model of phlegm-stasis cementation syndrome in Chinese mini-swine. After the operation, they were administered with drugs for 8 weeks. The SOD activity and MDA content of each group were observed at the 0th week (before the experiment), the 2nd week after the high-fat diet (before the operation or drug administration) , the 6th week after the high-fat diet (4 weeks after the drug administration) and the 10th week after the high-fat diet (8 weeks after the drug administration). Meanwhile, the myocardial enzymogram test and the HE staining pathological observation were performed at the end of the experiment. The changes in the myocardial cell ultra-structure were observed under transmission electron microscope. RESULT: Compared with the normal control group, the model group showed significant decrease in serum SOD activity and notable increase in MDA content from the 2nd week to the end of experiment (P < 0.05 and P < 0.01). In the 10th week, the CK, LDH and CK-MB levels in serum also significantly increased in the model group (P < 0.05 and P < 0.01), with obvious structural abnormality in myocardial tissue pathologic morphology and ultra-structure. Compared with the model group, TYTZ groups showed specific increase in serum SOD activity and oblivious decrease in the MDA level (P < 0.05 or P < 0.01). Meanwhile, TYTZ could significantly decrease serum CK and LDH levels in the model group (P < 0.05 or P < 0.01), attenuate the ischemia injury of myocardial tissue, and improve the ultra-structure of cardiomyocytes. CONCLUSION: TYTZ shows an obvious protective effect on the myocardial injury in Chinese mini-swine with coronary heart disease of phlegm-stasis cementation syndrome. Its mechanism is related to the resistance against free radical oxidation injury and the inhibition of the lipid per-oxidation.

[3.0 T magnetic resonance imaging of pancreatic cancer in an orthotopic nude murine model].

OBJECTIVE: To assess the methods of establishing an orthotopic nude murine model of human pancreatic cancer and explore the manifestations of MRI (magnetic resonance imaging) and its pathological fundamentals so as to provide research rationales for human pancreatic cancer. METHODS: The BXPC-3 cell of human pancreatic cancer was orthotopically planted in nude mice. And the animals were examined by a clinical 3.0 T magnetic resonance imager. The MRI examinations were analyzed along with their pathological findings. RESULTS: Among 18 mice, 15 of them grew tumors with a success ratio of 83.3%. The pathological findings conformed to those of high differentiation pancreatic parenchyma cancer. Comparing with the neighbor muscles, the tumors were homogeneous (66.7%, 10/15) or heterogeneous (33.3%, 5/15) of hypointense on T1-weighted images while homogeneous (26.7%, 4/15) or heterogeneous (73.3%, 11/15) hyperintense on T2-weighted images with heterogeneous enhancement. The border became obviously enhanced and there was mild central enhancement while the necrotic part showed no enhancement. CONCLUSION: 3.0T MRI can detect pancreatic neoplasms ≥ 2 mm and visualize clearly their locations, shapes, dimensions and internal structures in an orthotopic nude murine model. Thus it provides a visible framework for further studies of human pancreatic cancer.

[Roles of peroxiredoxin 6 in the regulation of oxidative stress to lipopolysaccharide-induced acute lung injury].

OBJECTIVE: To investigate the roles of peroxiredoxin (Prdx) 6 in the regulation of oxidative stress to lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. METHODS: Prdx6 knockout mice were tested for their genotype by PCR and Prdx6 protein expression was measured in lungs by immunohistochemistry. Eighteen male Prdx6 knockout mice were randomised to the Prdx6 knockout control group and the Prdx6 knockout LPS 24 h group, with 9 mice in each group. Eighteen male wild-type C57BL/6J mice were randomised to the wild-type control group and the wild-type LPS 24 h group, with 9 mice in each group. ALI was induced by intratracheal administration of 5mg/kg LPS. Twenty-four hours after stimulation, lung tissue slides were stained with hematoxylin and eosin for histological evaluation. The concentration of protein in the bronchial alveolar lavage fluid (BALF) was measured by using the Micro BCA Protein Assay Kit. Levels of reactive oxygen species (ROS) in the lungs were quantified by measurement of hydrogen peroxide (H(2)O(2)). Lipid and protein peroxidation were measured as levels of malondialdehyde (MDA) and protein carbonylation. H(2)O(2), MDA, protein carbonyl, total superoxide dismutase (SOD) activity, and total antioxidative capacity (TAOC) in lungs were measured by using assay kits from the manufacture. RESULTS: Prdx6 knockout mice presented their genotype and there was no Prdx6 protein expression in the lung. Increased polymorphonuclear cells in alveoli and bronchial wall thickening were observed in the lungs of LPS groups, which were more severe in Prdx6 knockout LPS 24 h group compared with wild-type LPS 24 h group. LPS instillation induced a significant elevated protein concentration in BALF in wild-type LPS 24 h group (441 ± 54) mg/L compared with wild-type control group (168 ± 20) mg/L (t = -4.71, P < 0.01). Significantly increased protein level in BALF was observed in Prdx6 knockout LPS 24 h group (770 ± 66) mg/L compared with wild-type LPS 24 h group (t = -3.69, P < 0.01). LPS instillation induced a significantly decreased SOD activity in wild-type LPS 24 h group (16.0 ± 1.2) U/mg protein compared with wild-type control group (26.5 ± 3.9) U/mg protein (t = 6.22, P < 0.01). SOD activity in Prdx6 knockout LPS 24 h group (14.5 ± 5.3) U/mg protein was not statistical different from wild-type LPS 24 h group (t = 0.56, P = 0.60). LPS instillation induced a significantly increased H(2)O(2) and MDA in wild-type LPS 24 h group [H(2)O(2)(52.3 ± 7.8) nmol/g protein; MDA (3.3 ± 0.5) nmol/mg protein] compared with wild-type control group [H(2)O(2)(29.5 ± 3.2) nmol/g protein, (t = -4.25, P < 0.01); MDA (1.6 ± 0.8) nmol/mg protein, (t = -5.94, P < 0.01)]. Significantly increased H(2)O(2) and MDA [H(2)O(2)(73.5 ± 12.4) nmol/g protein, (t = -3.01, P = 0.02); MDA (5.9 ± 0.9) nmol/mg protein, (t = -6.01, P < 0.01)] were observed in Prdx6 knockout LPS 24 h group compared with wild-type LPS 24 h group. No significant difference of protein carbonylation was observed in wild-type LPS 24 h group (6.9 ± 1.2) nmol/mg protein compared with wild-type control group (6.1 ± 0.9) nmol/mg protein (t = -1.62, P = 0.15). Significantly increased protein carbonylation (8.9 ± 0.9) nmol/g protein was observed in Prdx6 knockout LPS 24 h group compared with wild-type LPS 24 h group (t = -2.76, P = 0.03). LPS instillation induced a significantly decreased TAOC in wild-type LPS 24 h group (4.7 ± 0.6) U/mg protein compared with wild-type control group (6.5 ± 0.4) U/mg protein (t = 3.35, P < 0.01). Significantly decreased TAOC was observed in Prdx6 knockout LPS 24 h group (3.9 ± 0.4)U/mg protein compared with wild-type LPS 24 h group (t = 2.44, P = 0.04). The above parameters were not statistically different between Prdx6 knockout control group and wild-type control group. CONCLUSION: Deletion of peroxiredoxin 6 exaggerated LPS-induced acute lung injury with increased oxidative stress.

Primary alveolar soft part sarcoma of vertebra: a case report and literature review.

Alveolar soft part sarcoma (ASPS) is a rare malignant soft tissue tumor, which rarely occurs in bone. We present a case of ASPS in a 23-year-old man with a 2-month history of back pain. Computed tomography scanning and magnetic resonance images demonstrated a destructive process in the 12th thoracic vertebra associated with a unilateral soft tissue mass. The tumor showed evidence of hypervascularity on MRI; it obviously was enhanced on T1-weighted images after injection of Gd-GDPA, and signal voids were shown on all pulse sequences which may help to differentiate ASPS from other tumors of the vertebra. We believe that this is the first case of ASPS arising in a vertebra.

[Immunoregulatory role of 1, 25-dihydroxyvitamin D(3)-treated dendritic cells in allergic airway inflammation].

OBJECTIVE: To explore the mechanism and immunoregulatory role of 1, 25-dihydroxy vitamin D(3) [1, 25 (OH)(2)D(3)]-treated dendritic cells (DCs) in allergic airway inflammation. METHODS: Mouse bone marrow-derived DCs were treated by 1, 25 (OH)(2)D(3) for 72 h. The expression levels of different Notch ligands: Jagged1, Jagged2, Delta1, Delta3, and Delta4 in these DCs were detected by RT-PCR and Western blotting. Mouse spleen CD4+ T cells were cultured with 1, 25 (OH)(2)D(3)-treated DCs or 1, 25 (OH)(2)D(3)-treated DCs blocked by polyclonal antibody of Jagged1 or Jagged2 (control group) for 48 h. The percentage of CD4+CD25+Foxp3+ T cells in CD4+ T cells was detected by flow cytometry (FCM). Ten mice underwent intraperitoneal injection of ovalbumin (OVA) to be sensitized and then divided into 2 groups to inhale 1, 25 (OH)(2)D(3)-treated DC suspension and PBS-DC suspension respectively for 6 successive days. Then the mice were killed. Bronchoalveolar lavage fluid was obtained to detect the eosinophil count and the levels of interleukin (IL)-4, IL-6, IL-13, and interferon (IFN)-gamma, and pathological examination of lung was conducted. Spleens were taken out to isolate the CD4+ T cells, and immunolabeling and FCM were used to detect the percentage of CD4+CD25+Foxp3+ T cells. RESULTS: The mRNA and protein expression levels of Jagged1 and Jagged2 in the 1, 25 (OH)(2)D(3)-treated DCs were (0.376 +/- 0.029) and (0.786 +/- 0.034), and (0.564 +/- 0.018) and (0.632 +/- 0.026) respectively, all significantly higher than those of the control group [(0.146 +/- 0.032) and (0.124 +/- 0.025), and (0.267 +/- 0.012) and (0.098 +/- 0.012) respectively, all P < 0.01)]. The percentage of CD4+CD25+Foxp3+ T cells in the CD4+ T cells cultured with 1, 25 (OH)(2)D(3)-treated DCs was (22.49% +/- 0.56%), significantly higher than that of the a PBS control group [(6.67% +/- 0.60%), P < 0.01]. The percentage of CD4+CD25+Foxp3+ T cells in CD4+ T cells after cultured with 1, 25 (OH)(2)D(3)-treated DC blocked by polyclonal antibody of Jagged2 was (6.56% +/- 1.89%), significantly lower than that of the un-blocked control group [(20.37% +/- 1.64%), P < 0.01]. The levels of IL-4, IL-5, IL-13, and IFN-gamma, and eosinophil count in the BALF of the 1, 25 (OH)(2)D(3)-treated DC group were (33 +/- 5) pg/ml, (134 +/- 23) pg/ml, (91 +/- 11) pg/ml, and undetectable (< 12.5 pg/ml), and (236 +/- 29) x 10(3)/ml, all significantly lower than those of the PBS-DC group [(55 +/- 7) pg/ml, (332 +/- 49) pg/ml, (152 +/- 19) pg/ml, and (23 +/- 6) pg/ml, and (588 +/- 56) x 10(3)/ml, all P < 0.01]. The percentage of CD4+CD25+Foxp3+ T cells in the spleens of the 1, 25 (OH)(2)D(3)-treated DC group was (14.69% +/- 1.14%), significantly higher than that of the PBS-treated DC group [(2.38% +/- 0.14%, P < 0.01). CONCLUSION: Treatment of the DCs with 1, 25 (OH)(2)D(3) inhibits the allergic inflammation in the airway, maybe via the induction of CD4+CD25+Foxp3+ regulatory T cells by 1, 25 (OH)(2)D(3)-treated DCs through Jagged2-mediated Notch signal pathway.

[Clinicopathological study of 5 cases of proximal-type epithelioid sarcoma].

OBJECTIVE: To investigate the clinicopathological characteristics, diagnosis and differential diagnoses of proximal-type epithelioid sarcoma (PES). METHODS: Five cases of PES were retrieved from pathology files. Clinical, pathologic and immunohistochemical features of the tumors were reviewed. RESULTS: One patient was female and 4 were male. Ages of the patients ranged from 19 to 46 years. The sites of the tumor involvement were vulvar (2 cases), hypogastric zone (1 case), anterosuperior iliac spine (1 case) and buttock (1 case). Clinically, the tumor masses were painless and progressive solitary nodules. Microscopically, the tumor cell growth was infiltrative in nature, nodular in appearance with degenerative and necrotic cells at the central areas. The tumors consisted of relatively uniform epithelioid cells with round or oval nuclei and eosinophilic cytoplasm. Immunohistochemically, the tumor cells were positive for vimentin (5/5), CK (4/5), EMA (4/5), beta-catenin (3/5), CD34 (3/5), and S-100 protein (1/5), but were negative for SMA, MyoD1, Desmin, HMB-45, CK7 and CK20. CONCLUSION: Definitive diagnosis of PES relies on its histopathological characteristics in conjunction with appropriate immunohistochemical findings.

[An epidemiological study on HIV sexual transmission in married spouse in Yunnan province].

OBJECTIVE: To understand the status of HIV sexual transmission among HIV-sero-discordant spouses and HIV-sero-accordant spouses in Yunnan province, to discuss the related factors and to provide evidence for HIV prevention and control strategy. METHODS: Five places with serious epidemic and 3 moderate ones were voluntarily, randomly selected. According to time sequence, 300 spouses (600 people) with stable marriage were interviewed with questionnaire. RESULTS: HIV-sero-accordant spouses occupied for 40.7% of the total spouses under survey, with the others were HIV-sero-discordant ones. Among the ones that had already been diagnosed in the families, sexual transmission was their main mode of transmission, which was accounted for 68.3%, followed by IDU as 19.7%. After disclosed the HIV test outcomes to their spouses, 63.4% HIV-sero-discordant spouses and 47.0% HIV-sero-accordant ones changed their sexual behaviors. The rates of consistent condom use among the HIV-sero-discordant spouses increased from 16.8% to 95.0%, and in HIV-sero-accordant spouses increased from 8.2% to 60.9%. Data were analyzed by multi-factor logistic regression. Factors on influencing the sexual transmission in spouses would include condom use, frequency of sexual contacts and sexual transmission disease (STD) status etc. CONCLUSION: The main transmission mode for the first HIV infected spouse was sexual transmission. Factors influencing sexual transmission in spouses would include condom use, frequency of sexual contacts, STD situation and husband was the first one being infected in the families, etc. Disclosure of the HIV results to the spouses could make a significant changes in the frequencies of sexual contact as well as the rate of condom use.

OsSpo11-4, a rice homologue of the archaeal TopVIA protein, mediates double-strand DNA cleavage and interacts with OsTopVIB.

DNA topoisomerase VI from Archaea, a heterotetrameric complex composed of two TopVIA and two TopVIB subunits, is involved in altering DNA topology during replication, transcription and chromosome segregation by catalyzing DNA strand transfer through transient double-strand breaks. The sequenced yeast and animal genomes encode only one homologue of the archaeal TopVIA subunit, namely Spo11, and no homologue of the archaeal TopVIB subunit. In yeast, Spo11 is essential for initiating meiotic recombination and this function appears conserved among other eukaryotes. In contrast to yeast and animals, studies in Arabidopsis and rice have identified three Spo11/TopVIA homologues and one TopVIB homologue in plants. Here, we further identified two novel Spo11/TopVIA homologues (named OsSpo11-4 and OsSpo11-5, respectively) that exist just in the monocot model plant Oryza sativa, indicating that at least five Spo11/TopVIA homologues are present in the rice genome. To reveal the biochemical function of the two novel Spo11/TopVIA homologues, we first examined the interactions among OsSpo11-1, OsSpo11-4, OsSpo11-5, and OsTopVIB by yeast two-hybrid assay. The results showed that OsSpo11-4 and OsTopVIB can self-interact strongly and among the 3 examined OsSpo11 proteins, only OsSpo11-4 interacted with OsTopVIB. Pull-down assay confirmed the interaction between OsSpo11-4 and OsTopVIB, which indicates that OsSpo11-4 may interact with OsTopVIB in vivo. Further in vitro enzymatic analysis revealed that among the above 4 proteins, only OsSpo11-4 exhibited double-strand DNA cleavage activity and its enzymatic activity appears dependent on Mg(2+) and independent of OsTopVIB, despite its interaction with OsTopVIB. We further analyzed the biological function of OsSpo11-4 by RNA interference and found that down-regulated expression of OsSpo11-4 led to defects in male meiosis, indicating OsSpo11-4 is required for meiosis.