Objective To observe the effects of basic fibroblast growth factor (bFGF) on the expression of heat shock protein 70 (HSP70) in rat′s retina after ischemia/reperfusion injury. Methods The rat model of experimental retinal ischemia/ reperfusion injury was made by increasing the intraocular pressure. Tweenty-four Wistar rats were divided into normal (3 rats) and operation group (21 rats) randomly. The latter group was subdivided into group 0 hour, 4, 8, 12, 24, 48 and 72 hours after reperfusion, in which the left eyes of the rats were in the ischemia/reperfusion groups and the right ones were in the treatment groups (bFGF 2 ?g intracameral injection). The expression of HSP70 was observed by strept avidin-biotin complex (SABC) immunohistochemistry. Results No HSP70 positive cells were found in normal group; a few of HSP70 positive cells were found 0 hour after reperfusion [20.8?4.5)cells/mm 2], and increased gradually until reached the peak 24 hours later [(111.2?4.4) cells/mm 2] and then decreased gradually. Few HSP70 positive cells were found 72 hours after reperfusion. The amount of HSP70 positive cells increased in treatment group at all time courses, and the peak time was earlier and longer than that in ischemia group. HSP70 positive cells distributed extensively in retinal ganglion cell layer and inner nucleous layer. The difference of the amount of HSP70 positive cells between the two groups was significant ( P

Aim To study the characteristics of KCNQ2/3 potassium channel expressed in CHO cells and its modulation by M_1 receptor.Methods KCNQ2 and KCNQ3 potassium channels and M_1 receptor were co-expressed in CHO cells.Whole cell patch-clamp techniques was used to observe the characteristics of KCNQ2/3 current,its modulation by the M_1 receptor,and the effects of the common potassium channel blockers.Results KCNQ2/3 current recorded in CHO cells was a slow-activation low-threshold non-inactivating,voltage-dependent outward potassium current.KCNQ2/3 current was elicited at about-60 mV,V_(1/2)(-26.8?1.2) mV and the deactivation current fitted two exponential function,with ?_(fast) of 101ms and ?_(slow) of 309 ms.The channel was not sensitive to common pharmacological blockers such as 4-AP,Ba~(2+) and TEA,but was inhibited significantly by linopirdine,with a IC_(50) of(6.5?0.83) ?mol?L~(-1).Acetylcholine suppressed the KCNQ2/3 current reversibly via M_1 receptor,with a IC_(50) of(0.7?0.05) ?mol?L~(-1).Conclusion KCNQ2 and KCNQ3 channels are the molecular basis of M-current observed in neuronal cells.KCNQ2/Q3 current expressed in CHO cells has similar characteristics as that seen in neuronal M-current.Linopirdine is a powerful blocker of KCNQ2/3 channel and acetylcholine inhibits the current by muscarinic M_1 receptor.This experiment has laid a solid basis for further study of M-current and KCNQ2/3 current,and is important for the study of neurological diseases relating to alteration of M-current,such as convulsion,epilepsy and Alzheimers disease.

Aim To explore the relations between anti-apoptotic role of dipfluzine (DIP) and the death signaling transduction pathway initiated by CD95 molecules, and the transcription factor involved in the transcription regulation of CD95 molecules in the hippocampal CA1 region after transient forebrain ischemia. Methods The rat forebrain transient ischemia model was established through 15 min ischemia followed by 3 days reperfusion by using the four-vessel method. The rats were divided randomly into five groups: sham control group, ischemia / reperfusion (I/R) group, DIP treated groups (20, 40 and 80 mg·kg-1 body weight, ig, separately). Western blotting and RT-PCR were performed to detect the expression changes of Fas, FasL, caspase 10 p20, caspase 8, I-κB-α, and p-I-κB-α molecules in protein and mRNA levels, separately, and immunohistochemistry for molecular localization of Fas and FasL in rat hippocampus. Results The expression of Fas, FasL, and caspase 10 p20 in protein and mRNA levels increased after I/R, which was inhibited significantly after treatment with 20 and 40 mg·kg-1 of DIP (P<0.01). In 80 mg·kg-1 of DIP group, the expression of Fas and FasL protein was not significantly different from that of I/R group (P>0.05). The expression of caspase 8 and I-κB-α showed no significant differences in all groups (P>0.05), and no gene expression was observed for p-I-κB-α protein in the study. DIP significantly affected molecular distribution of Fas and FasL protein in CA1 subregion of hippocampus. Conclusion DIP inhibits the death signaling transduction pathway initiated by CD95 molecules in rat hippocampal CA1 subregion, and NF-κB transcription factor may not be involved in the transcription regulation of CD95 molecules after transient forebrain ischemia.

Objective To evaluate the accuracy of MSCT in the pre-operative N-staging and diagnosis of metastatic lymph nodes in each group of gastric cancer.Methods Pathological and CT data of 91 patients with gastric cancer proved by surgery and pathology were analyzed retrospectively.Three-phase dynamic enhancements were performed before surgery in a unified way of hypotonic oral water,N-stage and grouping of lymph nodes of the preoperative CT imaging were evaluated by using the established diagnostic criteria and then compared with the results of surgery and pathology,the accuracy of staging and grouping was analyzed by using Kappa test.Results The accuracy of MSCT diagnosing the N-staging as a whole was 86.3%.The accuracy for N0,N1,N2 and N3 was 83.5%,89.0%,83.5% and 89.0%,respectively.The sensitivity was 86.5%,83.3%,50% and 47.4%,respectively.The specificity was 79.5%,89.4%,89.6% and 100.0%, respectively.The sensitivity for N0 was statistically different from that for N2, N0 and N3(P≤0.007).The detected accuracy for the group of left side of the cardium (No.2), periphery of the splenic hilum (No.10), posterior of the pancreastic head (No.13) were higher than other groups on MSCT with the accuracyof 100%.The sensitivity for the group of No.2,periphery of the coeliac trunk(No.9),No.10,and No.13 was 100%.The specificity for the group of No.2,No.10,and No.13 was 98.9%.Conclusion Relatively high accuracy in the preoperative N-staging and diagnosis of metastatic lymph nodes in each group of gastric cancer can be obtained by MSCT, which provide reliable information for preoperative assessment and intraoperative lymph node dissections.

Chronic refractory wound refers to the wound with unclear etiology, multiple and complex injury factors, slow healing, and no obvious tendency of healing after treatment for 4 weeks. The formation and evolution process of chronic refractory wounds are very complex, involving re-epithelialization of wound tissue, cell proliferation, tissue remodeling, and angiogenesis and lymphangiogenesis. The abnormal expression of long non-coding RNA may be involved in the formation of chronic refractory wounds, but the specific pathogenesis and related molecular biological changes are still controversial. In this paper, we reviewed the process and role of long non-coding RNA in regulating keratinocyte differentiation, fibroblast proliferation, and regeneration of vascular and lymphatic endothelial cell in chronic refractory wounds.

This paper reviews recent work on biodegradable and bioabsorbable materials, including macromolecular, inorganic, and compound materials which have been used as bone-repaired devices. The properties and uses of poly(lactic acid), chitin and tricalcium phosphate are expounded. At the same time, we discuss the trend in the development of biodegradable and bioabsorbable bone-repaired materials. In our opinion, the development of biodegradable and bioabsorbable bone-repaired materials forcuses on the composite of different materials, especially the composite of BMP and MSCs; on the improvement of the mechanical properties of materials; and on the search for new suitable materials.
Absorbable Implants ; Bone Substitutes ; Calcium Phosphates ; Lactic Acid ; Materials Testing ; Polyesters ; Polymers

Objective:To observe the effect of immunofluorescence double staining for foamy macrophages and Mycobacterium tuberculosis (MTB) in paraffin-embedded tissue of clinical tuberculous wound, in comparison with three routine staining methods. Methods:The experimental method was used. From April 2019 to May 2020, 10 patients with tuberculous wound (5 males and 5 females, aged 28-77 years) meeting the inclusion criteria were treated in the Department of Burns and Plastic & Wound Repair Surgery of Xiang′an Hospital of Xiamen University. The paraffin-embedded wound tissue were collected during extended debridement and preserved in the Department of Pathology of this hospital. Forty paraffin sections were made from the wound tissue of each patient. Hematoxylin-eosin (HE) staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining were performed respectively, with 10 sections in each method. The section rejection rate of four staining methods were calculated. The recognition and detection of wound granuloma tissue in the four staining methods were observed and counted, and the recognition and detection of foamy macrophages in the wound tissue stained with four methods were observed. The MTB detection in the wound granuloma tissue and non-granuloma tissue in the four staining methods were compared. The subtyping and distribution of foamy macrophages and detection rate of MTB in the wound granuloma tissue and non-granuloma tissue, the morphologic clarity of foamy macrophages, as well as the non-specific staining rate and the loss rate of positive reaction of MTB and foamy macrophages by Ziehl-Neelsen and immunohistochemical double staining were compared with those of immunofluorescence double staining. Data were statistically analyzed with Fisher′s exact probability test, one-way analysis of variance, independent sample t test and Wilcoxon signed rank test. Results:The section rejection rate of HE staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, and immunofluorescence double staining were 3% (3/100), 1% (1/100), 6% (6/100), and 2% (2/100), respectively. There was no statistically significant difference among the four groups ( P=0.26). All the four staining methods could identify granuloma tissue, and the number of granuloma structures was similar ( F=1.284, P=0.28). All the four staining methods were able to identify foamy macrophages in the wound tissue, which was detected in each section. No MTB was observed in the wound granuloma tissue or non-granuloma tissue by HE staining or immunohistochemical staining. MTB was observed distributing in the wound granuloma tissue and non-granuloma tissue by Ziehl-Neelsen and immunohistochemical double staining and immunofluorescence double staining, and most MTB distributed in the wound granuloma tissue. Ziehl-Neelsen and immunohistochemical double staining could not distinguish foamy macrophages engulfed MTB from that non-engulfed MTB. Immunofluorescence double staining showed that foamy macrophages engulfed MTB mostly distributed in the wound granuloma tissue, and the foamy macrophages non-engulfed MTB mostly distributed in the wound non-granuloma tissue. The detection rates of MTB in wound granuloma and non-granuloma tissue in immunofluorescence double staining were (89.00±0.08)% and (82.67±0.05)%, respectively, which were significantly higher than (54.56±0.14)% and (44.44±0.13)% in Ziehl-Neelsen and immunohistochemical double staining ( t=-12.495, -7.961, P<0.01). Compared with that of Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining showed better foamy macrophages clarity in wound tissue ( Z=-3.162, P<0.01). The nonspecific staining rate and positive reaction loss rate of MTB and foamy macrophages in wound tissue of immunofluorescence double staining were (9.11±0.07)% and (9.22±0.07)%, respectively, which were significantly lower than (20.67±0.06)% and (44.00±0.12)% of Ziehl-Neelsen and immunohistochemical double staining ( t=4.569, 15.519, P<0.01). Conclusions:Compared with HE staining, immunohistochemical staining, and Ziehl-Neelsen and immunohistochemical double staining, the immunofluorescence double staining is easy to operate, giving clear and intuitive images. It allows accurate imaging co-localization of MTB and foamy macrophages in paraffin-embedded tissue of clinical tuberculous wound.

In this paper, poly(D,L-lactide) (PDLLA), MDI chain-extending poly(D,L-lactide) (PDLLA/MDI) and MDI chain-extending poly(D,L-lactide)/hydroxyapatite composite (PDLLA/HA/MDI) were prepared respectively and the effects of moulding and extruding conditions on their mechanical properties were also investigated. At the optimal conditions, bending strength of PDLLA and PDLLA/MDI is 35.1 MPa and 51.3 MPa, respectively, and their bending modulus is 2413.6 MPa and 1830.9 MPa, respectively. Bending strength of PDLLA/HA and PDLLA/HA/MDI is 31.2 MPa and 55.4 MPa, respectively, and their bending modulus is 1735.0 MPa and 2068.5 MPa, respectively. These results have shown that the mechanical properties of PDLLA/MDI and PDLLA/HA/MDI have enhanced significantly by MDI chain-extending.
Biocompatible Materials ; chemistry ; Durapatite ; chemistry ; Mechanics ; Medical Laboratory Science ; instrumentation ; methods ; Polyesters ; chemistry

Foamy macrophages (FM), also known as foam-like macrophages, refer to lipid-laden monocytes or macrophages. FM are a kind of inflammatory cells that are rich in lipid droplets in cytoplasm. In the diseases caused by Mycobacterium tuberculosis (Mtb), such as granuloma and tuberculous wounds, FM can not only inhibit the immune response, but also affect the prognosis and outcomes. The formation mechanisms of FM caused by Mtb infection have some specificity, which may be an important factor for its long-term survival in cells and influences on disease prognosis and outcomes. Therefore, studying the mechanisms of Mtb-mediated formation of FM is conductive to further reveal the pathological evolution of diseases and provide new ideas for further precise treatment. This article reviewed the mechanisms of Mtb-mediated formation of FM in recent years.

Objective:To explore the effects of Freund′s complete adjuvant on autophagy protein expression in rat tuberculous wound model.Methods:The experimental research method was used. In the first batch, twelve 6-week-old male Sprague-Dawley (SD) rats were sensitized by subcutaneous injection of Freund′s complete adjuvant into the hips. Three weeks later, the rats were infected with attenuated Bacille Calmette-Guérin (BCG) subcutaneously on both sides of the back spine. After establishing the tuberculosis wound rat model, according to the random number table (the same grouping method below), the rats were divided into 8 d infection group, 15 d infection group, 32 d infection group, and 43 d infection group, with 3 rats in each group, with continuous normal feeding to the corresponding days after infection. In the second batch, twenty-three 6-week-old male SD rats were divided into blank control group ( n=3, normal feeding without any treatment), BCG alone group ( n=5), BCG+ rapamycin group ( n=6), BCG+ 3-methyladenine group ( n=6), and BCG+ starvation group ( n=3). The last 4 groups of rats were sensitized as before, and infected as before 1 week later. Rats in BCG alone group were fed normally without any treatment. Rats in BCG+ rapamycin group or BCG+ 3-methyladenine group were intraperitoneally injected with rapamycin or 3-methyladenine once every other day and fed normally. Rats in BCG+ starvation group were fasted for 48 hours after infection and then fed normally. All the rats in the first batch of 4 groups were sacrificed on the corresponding days after infection, and the tissue where the buttocks were injected with Freund′s complete adjuvant was harvested; the tissue of rats in the second batch of BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group were harvested the same as before 7 days after infection, and all the rats in blank control group were taken the same tissue at the same time point. Hematoxylin-eosin staining was performed to observe the structure and morphology of cells in the tissue harvested; immunohistochemistry was used to observe the protein expressions of Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B) in the tissue harvested. Data were statistically analyzed with Kruskal-Wallis test and Bonferroni correction. Results:Inflammatory cell infiltration was observed in the tissue of rats where the Freund′s complete adjuvant was injected in 8 d infection group, granuloma formation was seen in 15 d infection group, part of tissue cell necrosis was seen in 32 d infection group and 43 d infection group, and cell necrosis in 43 d infection group was worse than that in 32 d infection group. Seven days after infection, inflammatory cell infiltration was seen in the tissue of rats where the Freund′s complete adjuvant was injected in BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group, while regular arrangement of cells and no inflammatory cell infiltration were observed in blank control group. There were no statistically significant differences in the protein expressions of Beclin-1 or LC3B in the tissue of rats where the Freund′s complete adjuvant was injected in 8 d infection group, 15 d infection group, 32 d infection group, and 43 d infection group ( H=1.923, 5.821, P>0.05). Seven days after infection, the protein expressions of Beclin-1 and LC3B in the tissue of rats where the Freund′s complete adjuvant was injected in blank control group, BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group were respectively 0.325% (0.250%, 0.360%), 3.225% (1.340%, 3.987%), 4.823% (2.630%, 6.559%), 4.216% (1.790%, 5.969%), 1.765% (0.865%, 2.649%), and 0.301% (0.264%, 0.516%), 2.865% (1.455%, 5.768%), 1.033% (0.398%, 1.873%), 1.168% (0.429%, 1.907%), 0.655% (0.283%, 1.652%). The protein expression of Beclin-1 in the tissue of rats where the Freund′s complete adjuvant was injected in BCG+ rapamycin group was significantly higher than that of blank control group ( Z=4.796, P<0.05). The protein expression of LC3B in the tissue of rats where the Freund′s complete adjuvant was injected in BCG alone group was significantly higher than that of blank control group ( Z=4.953, P<0.05). Conclusions:Freund′s complete adjuvant can enhance the expression levels of local tissue autophagy-related proteins Beclin-1 and LC3B in rat tuberculous wound model.