Objective: To investigate the levels of Th9 cells and interleukin-9 (IL-9), and to assess the regulatory activity of Th9/IL-9 to anti-tumor immune response in patients with gastric cancer. Methods: Thirty-four patients with gastric cancer who received operation in the First Affiliated Hospital of Xinxiang Medical University between October 2018 and August 2019 were included. Twenty individuals who received physical examination in the same period were also enrolled. Peripheral blood was collected, and then plasma and peripheral blood mononuclear cells (PBMCs) were isolated. Tumor-infiltrating lymphocytes (TILs) and autologous gastric cancer cells were isolated from resected gastric cancer tissues. CD4(+) T cells, CD8(+) T cells, and CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells were purified from PBMCs and TILs. Plasma IL-9 level was measured by enzyme linked immunosorbent assay (ELISA). The percentage of CD3(+) CD4(+) IL-9(+) Th9 cells in PBMCs and TILSs was assessed by flow cytometry. The mRNA levels of IL-9 and transcriptional factors purine-rich nucleic acid binding protein 1 (PU.1) were semi-quantified by real-time quantitative polymerase chain reaction (RT-qPCR). PBMCs and TILs from gastric cancer patients were stimulated with recombinant human IL-9. Cellular proliferation was measured by cell counting kit-8. The phosphorylation levels of signal transducer and activator of transcription 3 (STAT3) and STAT6 were investigated by western blot. Cytokine production was measured by ELISA. Purified CD8(+) T cells from TILs of gastric cancer patients were stimulated with recombinant human IL-9. CD8(+) T cells and autologous gastric cancer cells were cocultured in direct contact and indirect contact manner. The percentage of target cell death was calculated by measuring the lactate dehydrogenase (LDH) level. These cretion of γ-Interferon (γ-IFN) and tumor necrosis factor-α (TNF-α) was measured by ELISA. CD4(+) CCR4(-)CCR6(-)CXCR3(-)cells, CD8(+) T cells, and autologous gastric cancer cells were directly cocultured, and anti-IL-9 neutralizing antibody was added. The target cell death was measured. Results: The percentages of CD3(+) CD4(+) IL-9(+) Th9 cells in PBMCs of control group and PBMCs of gastric cancer group were (1.21±0.25)% and (1.14±0.19)%, respectively. The difference was not statistically significant (P=0.280). The percentage of CD3(+) CD4(+) IL-9(+) Th9 cells in TILs of gastric cancer group was (2.30±0.55)%, which was higher than those in PBMCs of control group and PBMCs of gastric cancer group (P<0.001). The plasma IL-9 level in control group and gastric cancer group were (5.04±1.51) and (4.93±1.25) ng/ml. The difference was not statistically significant (P=0.787). The relative levels of IL-9 mRNA in PBMCs of control group and PBMCs of gastric cancer group were 1.33±0.39 and 1.36±0.27. The difference was not statistically significant (P=0.691). The relative level of IL-9 mRNA in TILs of gastric cancer group was 2.90±0.75, which was higher than those in PBMCs of control group (P<0.001) and PBMCs of gastric cancer group (P<0.001). The relative levels of PU.1 mRNA in PBMCs of control group and PBMCs of gastric cancer group were 1.21±0.12 and 1.20±0.11. The difference was not statistically significant (t=0.21, P=0.833). PU.1 mRNA relative level in TILs of gastric cancer group was 2.81±0.65, which was higher than those in PBMCs of control group (P<0.001) and PBMCs of gastric cancer group (P<0.001). Recombinant human IL-9 stimulation did not affect the proliferation of PBMCs and TILs of gastric cancer patients (P>0.05), but elevated the phosphorylation level of STAT6 and induced the secretions of γ-IFN, IL-17, and IL-22 by TILs (P<0.05). In direct contact culture system, IL-9 stimulation promoted tumor-infiltrating CD8(+) T cells-induced autologous gastric cancer cell death [(20.62±2.27)% vs. (16.08±2.61)%, P<0.01)]. In indirect contact culture system, IL-9 stimulation did not increase CD8(+) T cell-induced autologous gastric cancer cell death [(5.21±0.70)% vs. (5.31±1.22)%, P=0.998)]. However, the secretion levels of γ-IFN were elevated in response to IL-9 stimulation in both culture systems [direct contact culture system: (100.40±12.05) pg/ml vs. (76.45±8.56) pg/ml; indirect contact culture system: (78.00±9.98) pg/ml vs. (42.09±10.71) pg/ml; P<0.01]. The TNF-α secretion level did not significantly changed (P>0.05). In direct contact culture system, the percentage of target cells was (22.01±3.05) % and γ-IFN secretion level was (104.5±12.84) pg/ml in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells group, which was higher than (16.08±2.61)% and (76.45±8.56) pg/ml in CD8(+) T cells+ gastric cancer cells group (P<0.01). However, the percentage of target cells was (14.47±3.14)% and γ-IFN secretion level was (70.45±19.43) pg/ml in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells+ anti-IL-9 neutralizing antibody group, which were lower than those in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells group (P<0.01). Conclusion: Tumor-infiltrating Th9 cells and the secreting IL-9 promote the activity of CD8(+) T cells in gastric cancer patients, and enhance anti-tumor immune response.
Humans ; CD8-Positive T-Lymphocytes ; Stomach Neoplasms/pathology* ; Tumor Necrosis Factor-alpha/metabolism* ; Lymphocytes, Tumor-Infiltrating/pathology* ; Interferon-gamma/metabolism* ; RNA, Messenger/metabolism* ; Antibodies, Neutralizing/metabolism*

OBJECTIVE: To observe the clinical effect of grain-moxibustion combined with medicine therapy for asthenospermia and oligospermia. METHODS: A tatal of 60 patients were randomized into an observation group (30 cases) and a control group (30 cases) according to 1︰1 ratio. In the control group, vitamin E capsules were taken orally one capsule each time, twice a day, and pills 6 g each time, three times a day for a total of 3 months. In the observation group, grain-moxibustion was applied at Guanyuan (CV 4)，Shenshu (BL 23) and Zusanli (ST 36) based on the control group, once a week for 3 months, with a total of 12 times. The sperm concentration and sperm progressive motility were measured by automatic sperm quality analysis system in the two groups, and the clinical effects were compared. Sperm DNA fragmentation index (DFI) in the observation group was measured by sperm nucleus chromosome structure assay (SCSA). RESULTS: ①The sperm concentrations and sperm progressive motilities after 1-month, 2-month and 3-month of treatment were increased compared with those before treatment in the two groups (<0.01), and they were increased with time. In the two groups, 2-month and 1-month of treatment, 3-month and 2-month of treatment were compared, the sperm concentrations and sperm progressive motilities were significantly increased (<0.01). The sperm concentrations after 1-month, 2-month and 3-month of treatment in the observation group were higher than those in the control group (<0.01), the sperm progressive motility after 3-month of treatment in the observation group was higher than that in the control group (<0.05). ②After 3-month of treatment，the DFI in the observation group was significantly reduced compared with that before treatment (<0.01). ③The total effective rate in the observation group after 3-month of treatment was 86.7% (26/30), which was superior to 63.3% (19/30) in the control group (<0.05). CONCLUSION Grain-moxibustion combined with medicine therapy can improve sperm concentration and sperm progressive motility, enhance the integrity of sperm DNA.
Humans ; Male ; Medicine, Chinese Traditional ; Moxibustion ; Oligospermia ; therapy ; Sperm Count ; Sperm Motility ; Spermatozoa

Objective To evaluate the efficacy and safety of combined use of ticagrelor and cilostazol for patients with acute coronary syndrome (ACS) complicated with upper digestive tract diseases following percutaneous coronary intervention ( PCI).Methods A total of 262 consecutive ACS patients complicated with upper digestive tract diseases followed-up for one-year after PCI were included in this study.The patients were allocated into control group (combined use of ticagrelor and aspirin , n＝184) and cilostazol group ( combined use of ticagrelor and cilostazol , n ＝78) for antiplatelet treatment.The basic characteristics of the patients , change of the treatment regimens , cardiovascular events and hemorrhagic events were compared between two groups .Results After one year of follow-up, 16.8%(31/184)patients in control group and 3.8%(3/78)in cilostazol group changed antiplatelet regimens (χ2＝8.200,P＝0.004).There was no statistical difference in use of statins and ACEI/ARB between two groups(P＞0.05).The rate of proton pump inhibitor use in control group was significantly higher than that in cilostazol group [82.1%(151/184) vs.52.6%(41/78), χ2＝24.35, P＝0.000].However, the dosage of β-blockers in cilostazol group was significantly higher than that in control group [(39.1 ±12.4) mg vs.(28.6 ±10.1) mg, t ＝7.174,P＝0.000].No statistical difference was found in total cardiovascular events between two groups [21.7%(40/184) vs.12.8%(10/78),χ2＝2.822,P＝0.121].The incidence of gastrointestinal hemorrhage in control group was significantly increased compared with cilostazol group [12.0%(22/184) vs.2.6%(2/78),χ2＝5.807,P ＝0.018], however, there was no significant difference in hemorrhagic events concerning the thrombolysis for myocardial infarction between two groups [17.4%(32/184) vs.9.0%(7/78), χ2＝3.063,P＝0.089].Conclusion Combined use of cilostazol and ticagrelor is effective and safe for ACS patients with gastrointestinal hemorrhage or a higher risk of hemorrhage .

Aim To primarily discuss the mechanism of Rhizoma Alismatis decoction on prevention and treatment of hyperlipemia,through investigating hepat-ic expressions of peroxisome proliferator activated re-ceptor-α(PPARα)and acyl CoA oxidase (ACO)in rats with hyperlipemia. Methods Fifty male Sprague-Dawley rats(160 ~ 180 g)were randomly divided into five groups:control group,model group,high-dose of Rhizoma Alismatis decoction group (H-RAD),low-dose of Rhizoma Alismatis decoction group(L-RAD), and Xue-Zhi-Kang group(XZK). Rats in control group were fed with ordinary forage,and the others were with high-fat forage,which lasted for four weeks. At the same time,high and low-dose of Rhizoma Alismatis decoction,as well as Xuezhikang capsule,was admin-istered in respectively designed groups. Then,the TC, TG,HDL and LDL of serum were detected,the mor-phology of liver tissues was observed with HE,and the expressions of PPARα and ACO were detected with RT-PCR and Western blot after four weeks. Results Rhizoma Alismatis decoction could significantly reduce serum concentration of TC,TG and LDL(P < 0. 01), while increasing the concentration of HDL(P < 0. 01) and strengthening the expressions of PPARα and ACO (P < 0. 01). Conclusion Strengthening the expres-sions of PPARα and ACO can be viewed as mecha-nisms of Rhizoma Alismatis decoction in prevention and treatment of hyperlipemia.

OBJECTIVE To observe the effect of electroacupuncture on the expression of VEGF and VEGFR2 in the uterusof the rat model of ovulation induction and to investigate the possible mechanism.METHODS Female SD rats aged 10 weekswere divided into control,model and treat groups randomly,10 in each group.The model group and the treatment group wereperitoneal injected with pregnant horse serum gonadotrophin(PMSG)of 20 U and injected of the same dose of human chorionicgonadotropin(hCG)after 48 h,and then cohabitated with male SD nice until next morning.A week before cohabitation,thetreat group received a consecutive electroacupuncture treatment for 7 d(dilatational wave of 2/15 Hz for 30 min and stimulativeintensity of 1 mA).The animals were sacrificed 3.5 d after the cohabitation.The uterine tissue was taken to observe the morphologicalchange,immunehistochemical method was used to inspect the expression of VEGF and VEGFR2,and qPCR technologywas used to determine the expression of VEGF and VEGFR,with the calculation by 2-△△Ct .RESULTS Macroscopicobservation:the surface in control group was regular and smooth;the model group was intumescentia,dull-red and hyperaemia;the treatment group was slightly swollen,rosy and no congestion.The expression of VEGF and VEGFR2 were observed inthe 3 groups by immunohistochemical method,with no significant difference in average optical density,but differences in theexpression distribution.Compared with the control group,the expression distribution of stromal cells and glands in the modelgroup was more extensive,and the expression of VEGFR2 in myometrium increased significantly(P <0.05).The electroacupuncturetreatment inhibited the the expression of VEGFR2 significantly(P <0.05).The expression of VEGF mRNA in uterinetissues of model group was 0.68 times of control group,with significant declining trend;the treat group was 0.88 times ofmodel group,with significant increasing trend.The expression of VEGF2 mRNA in uterine tissues of model group was 0.46times of control group,with significant declining trend;the treat group was 0.82 times of model group,with significant increasingtrend.CONCLUSION The electroacupuncture can adjust the expression of VEGF and its receptor in the uterine tissue of ovulation induction rat model,thus regulate the synchronization of the endometrial planting window caused by hormone

BACKGROUNDGlobally, the proportion of child deaths that occur in the neonatal period remains a high level of 37-41%. Differences of cause in neonate death exist in different regions as well as in different economic development countries. The specific aim of this study was to investigate the causes, characteristics, and differences of death in neonates during hospitalization in the tertiary Neonatal Intensive Care Unit (NICU) of China.

METHODSAll the dead neonates admitted to 26 NICUs were included between January l, 2011, and December 31, 2011. All the data were collected retrospectively from clinical records by a designed questionnaire. Data collected from each NICU were delivered to the leading institution where the results were analyzed.

RESULTSA total of 744 newborns died during the 1-year survey, accounting for 1.2% of all the neonates admitted to 26 NICUs and 37.6% of all the deaths in children under 5 years of age in these hospitals. Preterm neonate death accounted for 59.3% of all the death. The leading causes of death in preterm and term infants were pulmonary disease and infection, respectively. In early neonate period, pulmonary diseases (56.5%) occupied the largest proportion of preterm deaths while infection (27%) and neurologic diseases (22%) were the two main causes of term deaths. In late neonate period, infection was the leading cause of both preterm and term neonate deaths. About two-thirds of neonate death occurred after medical care withdrawal. Of the cases who might survive if receiving continuing treatment, parents' concern about the long-term outcomes was the main reason of medical care withdrawal.

CONCLUSIONSNeonate death still accounts for a high proportion of all the deaths in children under 5 years of age. Our study showed the majority of neonate death occurred in preterm infants. Cause of death varied with the age of death and gestational age. Accurate and prompt evaluation of the long-term outcomes should be carried out to guide the critical decision.

Cause of Death ; China ; Female ; Hospital Mortality ; Humans ; Infant ; Infant Mortality ; Infant, Newborn ; Infant, Newborn, Diseases ; mortality ; Intensive Care Units, Neonatal ; statistics & numerical data ; Male ; Perinatal Death ; Retrospective Studies

OBJECTIVETo investigate the blood supply of the pedicle fat grafts with the third lumbar segmental artery and its clinical effects on reoperation for lumbar disc herniation.

METHODSTwelve sides of 6 adult cadaver examples were contributed to investigate the courser of lumbar segmental vessels and the distribution of hypodermic capillary net of the dorsal branch of the third lumbar segmental artery. From January 2000 to January 2007,49 patients needed reoperation to treat lumbar disc herniation,including 26 males and 23 females with an average age of 55.6 years (ranged from 39 to 70 years). Duration between two operations ranged from 8 months to 15 years with an average of 6.9 years. Reoperative reasons included recurrent lumbar disc protrusion(30 cases)postoperative epidural scar formation (17 cases), postoperative epidural cyst formation (2 cases). Of them,9 patients underwent posterior lumbar interbody fusion at the second operation. The pedicle fat grafts with the third lumbar segmental artery were covered on the sites of the laminectomy in these patients. After negative pressure drainage tube were pulled out, 2 ml Chitsan were injected to the sites of the laminectomy and around epidural nerve root through epidural catheter. VAS score and the Oswestry Disability Index (ODI) were used to assess clinical outcomes before and after operation.

RESULTSThe courser of third lumbar segmental vessels were invariant at the lateral face of the lumbar vertebral body. The dorsal branch of the third lumbar segmental artery penetrated thoracolumbar fascia and formed rich hypodermic capillary net in the region. All patients were followed up from 5 to 8 years with an average of 5.6 years. VAS score of low back pain and leg pain decreased respectively from preoperative 7.6 +/- 1.2, 8.9 +/- 0.9 to 3.6 +/- 0.5, 3.0 +/- 0.4 at final follow-up (P < 0.01); and ODI score decreased from preoperative 44.1 +/- 6.2 to 13.9 +/- 3.6 at final follow-up (P < 0.01). According to ODI score to evaluate the clinical outcomes, 29 cases got excellent results, 11 good, 7 fair, 2 poor.

CONCLUSIONThe pedicle fat grafts with the third lumbar segmental artery and Chitsan can reduce epidural scar formation and prevent peridural fibrosis and adhesion and improve clinical effects of reoperation for lumbar disc herniation.

Adipose Tissue ; pathology ; Adult ; Aged ; Arteries ; pathology ; physiopathology ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Displacement ; pathology ; physiopathology ; surgery ; Lumbar Vertebrae ; blood supply ; pathology ; surgery ; Male ; Middle Aged ; Reoperation ; Transplantation ; Treatment Outcome

OBJECTIVETo screen genes related to peritoneal metastasis of colorectal cancer.

METHODSSpecimens of primary cancer and normal mucosa tissues were collected from 3 patients with peritoneal metastasis of colorectal cancer. The total RNA were extracted and inversely transcribed into cDNA to synthesize aRNA using in vitro RNA synthesis. The synthesized aRNA, after labeling with Cy3, were hybridized with the whole human genome oligo microarray. The Empirical Bayes method was used to screen the differentially expressed genes, followed by confirmation of the selected genes by semi-quantitative RT-PCR.

RESULTSWith a threshold of P≤0.05, a total of 105 differentially expressed genes were identified in primary cancer lesions, including 42 up-regulated and 63 down-regulated genes. Three of the up-regulated genes (S100P, PRDX1 and SLPI) were selected and confirmed by RT-PCR, which yielded results consistent with those from gene microarray.

CONCLUSIONGene microarray technique can provide valuable clues for locating the tumor markers of peritoneal metastasis in colorectal cancer patients.

Adenocarcinoma ; genetics ; secondary ; Adenocarcinoma, Mucinous ; genetics ; secondary ; Adult ; Aged ; Calcium-Binding Proteins ; genetics ; metabolism ; Colorectal Neoplasms ; genetics ; pathology ; Female ; Gene Expression Profiling ; Genome-Wide Association Study ; Humans ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; metabolism ; Oligonucleotide Array Sequence Analysis ; methods ; Peritoneal Neoplasms ; genetics ; secondary ; Peroxiredoxins ; genetics ; metabolism ; Secretory Leukocyte Peptidase Inhibitor ; genetics ; metabolism

Clinical characteristics,including primary and secondary sexual characteristics,basal endocrine profiles,and imaging results were reviewed.Follow-up data were recorded.16 patients had normal karyotypes,manifest amastia,infantile genitalia,amenorrhea,and delayed epiphyseal fusion at the knee and wrist joints.Serum gonadotropic hormone levels were significantly below normal values.15 patients underwent a gonadotropin-releasing hormone (GnRH) stimulation test and 6 patients had a prolonged GnRH stimulation test.16 patients underwent pituitary or brain magnetic resonance imaging (MRI),which showed small pituitaries in three patients,wing tips of suspicious nodules in 2 patients,an empty sella turcica in 1 patient,and a missing right olfactory bulb and tract in 1 patient.1 patient had no detectable uterus or accessory organs,while the other patients had primordial uteri.1 patient was diagnosed as a case of severe osteoporosis.1 patient suffered from pituitary stalk interruption syndrome.An artificial menstrual cycle due to hormone replacement therapy was not sustained after discontinuation of hormone therapy.As disease severity and the date of initiating hormone replacement varied,the results of treatment were quite different.For patients of reproductive age,it was rare to see a reversal of idiopathic hypogonadotropic hypogonadism after discontinuation of hormone therapy.

BACKGROUNDPoly (ADP-ribose) polymerase (PARP) plays an important role in cell survival and death. However, the mechanisms involved are not fully understood. Therefore, we investigated the effect of inhibition of PARP on acute myocardial infarction (AMI) at different time points in rats.

METHODSAMI was induced in rats by ligating the left anterior descending coronary artery. One group received 3-aminobenzamide (3-AB, a kind of PARP inhibitor) (30 mg/kg) by intraperitoneal injection. The changes of ultramicrostructure of cardiocytes in infarction region were noted, PARP cleavage was measured by Western blotting, and expressions of protein of PARP and apoptosis inducing factor (AIF) were measured by immunohistochemical staining after treatment with 3-AB for 2 hours, 4 hours, 6 hours, 1 week, 4 weeks and 8 weeks.

RESULTSFew damages to the ultramicrostructure of cardiocytes were observed after treatment with 3-AB. PARP cleavage was detected as early as 4 hours and markedly increased by 6 hours following AMI without 3-AB, but was not found until 6 hours following AMI treated with 3-AB. There were significant differences between 3-AB and AMI groups at the same time points. The expression of PARP was observed gradually increased, but that of AIF was suppressed for 6 hours after treatment of 3-AB, compared with AMI groups in positive cells at the same time points. There was significantly less cleavage of PARP and more PARP expression in 3-AB treated group compared with AMI and control groups at all matched time points.

CONCLUSIONSOur results suggest that 3-AB inhibits degradation of PARP, increases the expression of PARP protein, and suppresses the expression of AIF protein. Inhibition of PARP activity may protect cardiocytes in rats with AMI and reduce apoptosis.

Animals ; Apoptosis Inducing Factor ; metabolism ; Benzamides ; pharmacokinetics ; Blotting, Western ; Enzyme Inhibitors ; pharmacology ; Immunohistochemistry ; Male ; Myocardial Infarction ; metabolism ; prevention & control ; Myocytes, Cardiac ; drug effects ; metabolism ; Poly(ADP-ribose) Polymerases ; metabolism ; Rats ; Rats, Wistar