[Accuracy and clinical efficacy of robot-assisted transiliac-transsacral screw placement for posterior pelvic ring fractures].

OBJECTIVE: To explore accuracy and clinical effect of robot-assisted implantation of sacroiliac penetrating screw in orthopedic surgery for posterior pelvic ring fracture. METHODS: The clinical data of 24 patients with posterior pelvic ring fracture treated with robot-assisted sacroiliac penetration screws from August 2022 to August 2023 were retrospectively analyzed, including 10 males and 14 females; aged from 21 to 73 years old with an average of (49.29±14.48) years old;according to Tile pelvic fractures, 13 patients were type B and 11 were type C. The effect of screw placement was evaluated according to Gras criteria based on postoperative CT scan results. At the final follow-up, fracture healing was evaluated according to Matta score, and functional recovery was evaluated by Majeed score. RESULTS: All patients were followed up for 3 to 13 months with an average of (6.00±3.28) months. Totally 36 sacroiliac penetrating screws, 18 S1 penetrating screws, 18 S2 penetrating screws were inserted, a total of 29 were excellent and 7 good according to Gras standard. Screw adjustment times was 0.00 (0.00, 0.75) times. At the final follow-up, Matta score was excellent in 18 patients, 5 good and 1 moderate, and the maximum displacement distance was 2.55 (0.00, 5.65) mm. Majeed score was 84.37±8.38, 15 patients were excellent, 7 good and 2 moderate. CONCLUSION: Robot could accurately and safely assist in the placement of sacroiliac joint screws for the treatment of posterior pelvic ring fractures, and promote postoperative functional recovery of patients.

Serum chitinase-3-like protein 1 is a biomarker of liver fibrosis in patients with chronic hepatitis B in China.

BACKGROUND: Serum chitinase-3-like protein 1 (CHI3L1) is a potential biomarker for fibrosis assessment. We aimed to evaluate serum CHI3L1 as a noninvasive diagnostic marker for chronic hepatitis B virus-related fibrosis. METHODS: Serum CHI3L1 levels were measured by ELISA in 134 chronic hepatitis B (CHB) patients. Significant fibrosis was defined as a liver stiffness > 9.7 kPa. The performance of CHI3L1 was assessed and compared to that of other noninvasive tests by receiver operating characteristic (ROC) analysis. RESULTS: Serum CHI3L1 levels were significantly higher in CHB patients with significant hepatic fibrosis (≥ F2, 81.9 ng/mL) than in those without significant hepatic fibrosis (< F2, 56.5 ng/mL) (P < 0.001). In CHB patients, the specificity and sensitivity of CHI3L1 for predicting significant fibrosis were 75.6% and 59.1%, respectively, with a cut-off of 76.0 ng/mL and an area under the ROC curve of 0.728 (95% CI: 0.637-0.820). CONCLUSIONS: Serum CHI3L1 levels could be an effective new serological biomarker for the diagnosis of liver. Moreover, CHI3L1 is feasible in monitoring disease progression.

Acellular ostrich corneal stroma used as scaffold for construction of tissue-engineered cornea.

AIM: To assess acellular ostrich corneal matrix used as a scaffold to reconstruct a damaged cornea. METHODS: A hypertonic saline solution combined with a digestion method was used to decellularize the ostrich cornea. The microstructure of the acellular corneal matrix was observed by transmission electron microscopy (TEM) and hematoxylin and eosin (H&E) staining. The mechanical properties were detected by a rheometer and a tension machine. The acellular corneal matrix was also transplanted into a rabbit cornea and cytokeratin 3 was used to check the immune phenotype. RESULTS: The microstructure and mechanical properties of the ostrich cornea were well preserved after the decellularization process. In vitro, the methyl thiazolyl tetrazolium results revealed that extracts of the acellular ostrich corneas (AOCs) had no inhibitory effects on the proliferation of the corneal epithelial or endothelial cells or on the keratocytes. The rabbit lamellar keratoplasty showed that the transplanted AOCs were transparent and completely incorporated into the host cornea while corneal turbidity and graft dissolution occurred in the acellular porcine cornea (APC) transplantation. The phenotype of the reconstructed cornea was similar to a normal rabbit cornea with a high expression of cytokeratin 3 in the superficial epithelial cell layer. CONCLUSION: We first used AOCs as scaffolds to reconstruct damaged corneas. Compared with porcine corneas, the anatomical structures of ostrich corneas are closer to those of human corneas. In accordance with the principle that structure determines function, a xenograft lamellar keratoplasty also confirmed that the AOC transplantation generated a superior outcome compared to that of the APC graft.

[Raman spectra of single human living erythrocyte with the effect of pH and serum albumin].

In the present work, a cell environment which mimicked the real body environment according to the concentration radio between serum albumin and hemoglobin was built, and the cell morphology, the membrane deformation capacity, and the structure of intracellular hemoglobin of single human living erythrocyte under the effect of pH and serum albumin were studied. It was found that at different suspension pH, the magnitude of variations in cell shape and membrane deformation capacity changes with the structural changes of the intracellular hemoglobin. At pH 4. 14, 4. 76 and 10. 18, the loss of helical structure for hemoglobin, exposing of the hydrophobic amino acid in the globin chains, and changing of the combination of heme and globin, would completely destroy the stability of hemoglobin's structure, which seriously changes RBC's morphology and membrane deformation capacity. While at pH 6. 51 and 7. 80, the Raman spectra of erythrocytes are found to have no such changes, indicating that the structure of intracellular hemoglobin was not varied, thus the cell morphology and membrane deformation capacity are quite close to the normal values. At pH 5. 49 and 8. 76, RBC's morphology and membrane deformation capacity have different degrees of variation, but the structure of intracellular hemoglobin has not changed, suggesting that the cell morphology and membrane deformation capacity may be reversible. The results suggest that in the suspension solution containing serum albumin, erythrocytes have better ability to regulate and control the variation of the extracellular pH. In summary, upon building an environment which contains the same concentration radio of serum albumin to hemoglobin in the blood, this work performed systematic studies on the effect of pH on human erythrocytes. It can not only help to solve the problems about the mechanism of the structural and functional changes of erythrocytes induced by environmental pH, but also elucidates the possible variation of the erythrocyte under different pathologic states. It is therefore significant for the prevention and treatment of some diseases.

Pathway and mechanism of pH dependent human hemoglobin tetramer-dimer-monomer dissociations.

Hemoglobin dissociation is of great interest in protein process and clinical medicine as well as in artificial blood research. However, the pathway and mechanisms of pH-dependent human Hb dissociation are not clear, whether Hb would really dissociate into monomers is still a question. Therefore, we have conducted a multi-technique investigation on the structure and function of human Hb versus pH. Here we demonstrate that tetramer hemoglobin can easily dissociate into dimer in abnormal pH and the tetramer → dimer dissociation is reversible if pH returns to normal physiological value. When the environmental pH becomes more acidic (<6.5) or alkaline (>8.0), Hb can further dissociate from dimer to monomer. The proportion of monomers increases while the fraction of dimers decreases as pH declines from 6.2 to 5.4. The dimer → monomer dissociation is accompanied with series changes of protein structure thus it is an irreversible process. The structural changes in the dissociated Hbs result in some loss of their functions. Both the Hb dimer and monomer cannot adequately carry and release oxygen to the tissues in circulation. These findings provide a comprehensive understanding on the pH-dependent protein transitions of human Hb, give guideline to explain complex protein processes and the means to control protein dissociation or re-association reaction. They are also of practical value in clinical medicine, blood preservation and blood substitute development.

[Raman spectra of different kinds of thalassemia erythrocytes with the effect of pH].

Thalassemia is a kind of blood diseases which has high morbidity and large influence. Previous methods for diagnosis are all very cumbersome and time consuming. By comparing Raman spectra of different kinds of thalassemia and normal erythrocytes at acid or alkaline pH, it was found that beta-thalassemia and alpha-thalassemia erythrocytes have dissimilar Raman spectra in the acidic environment, such as the Raman spectra of beta-thalassemia erythrocytes showed higher intensity at the characteristic bands assigned to oxyhemoglobin, and the characteristic bands assigned to deoxyhemoglobin were even completely replaced; beta-thalassemia erythrocytes membrane has a smaller chain interaction between the transverse order parameters than normal erythrocytes, and the S(lat) values are different for different stages of anemia, while the S(lat) values are similar between alpha-thalassemia and normal erythrocytes, indicating that based on the effect of pH it is possible to diagnose thalassemia more quickly by using Raman spectra.

Characteristic variation of α-fetoprotein DNA nanometer-range irradiated by iodine-125.

To obtain the characteristic variation of structure and functional groups of α-fetoprotein (AFP) DNA irradiated by iodine-125((125)I), the AFP antisense oligonucleotide labeled with various radioactivity dose (125)I was mixed with the AFP DNA in a simulated polymerase chain reaction temperature condition. After the mixtures were irradiated by the (125)I from 2 to 72 hours, the mutation of the biogenic conformation and functional groups of the irradiated DNA were investigated using laser Raman spectroscopy. The shifted peak and the decreased intensity of the characteristic Raman spectra were found, which demonstrated that the structure of the phosphodiester linkage was broke, the pyridine and purine bases in DNA emerged and damaged. The model of gene conformation changed from form B to form C spectrum after the nanometer-range irradiation with (125)I from 2 to 24 hours. The damage of local pyridine and purine bases gradually increased along with the accumulation of irradiation, and the bases and ribosome were finally dissociated and stacked.

[Significance of preoperative screening of deep vein thrombosis on the prevention from pulmonary embolism].

OBJECTIVE: To Explore the significance of preoperative screening of deep vein thrombosis (DVT) on the prevention from pulmonary embolism (PE) with color Doppler flow imaging (CDFI) in patients of lower limb fractures. METHODS: A total of 2 000 patients with lower limb fractures were retrospectively analyzed from January to September in 2010. All the patients were scheduled to be operated. There were 1 140 males and 860 females, ranging in age from 18 to 94 years, with a mean of (54.78 +/- 21.45) years. Swollen limbs could be observed in all patients when admitted and traditional Chinese medicine therapies were given by both internal and external administrations. Selective internal or external fixations were scheduled 3 to 14 days after injured and the swell relieved. CDFI screenings for DVT were applied to all the patients one day before operations and clinical data, such as gender, age, fracture site and course of diseases, were analyzed. RESULTS: Of all the 2 000 patients, 128 patients had lower limbs DVTs (6.4%). Among all the DVT patients, 52 patients had DVT within vena iliacas or femoral veins,20 patients had DVT within popliteal veins or vena crualis, 56 patients had DVT within small muscular veins of legs. Thrombolytic therapies or inferior vena cava filters were taken to 72 patients with DVTs proximal to vena cruralises and thrombi disappeared in 20 cases. Inferior vena cava filters were placed in 17 patients. There were 28 patients with either failed thrombolytic or no filters placement. All above 65 patients were treated with fracture reduction, and other 7 patients were treated with conservative methods. Of all the operated patients, one patient had PE during operation and recovered after immediately rescue. Among 56 patients with DVT within small muscular veins of legs, 51 patients were treated with reduction without other special treatment, and other 5 patients were treated with conservative methods. There was no relationship between DVT occurrence and gender. The youngest sufferer was 22 years old. DVT occurrences of patients over 41 years old were significantly higher than that of patients under 40 years old. Higher occurrences were also observed in patients with multiple fractures, fracture of femurs compared with those with tibia or fibula fractures. DVT was found as early as 3 days after injury in one case. The longer the time from injuries to operations, the higher the occurrences of DVTs. CONCLUSION: Routine CDFI screenings should be applied to traumatic fractured patients before operations no matter whether there are DVTs existing or not. That is very important for the prevention of fatal PE during both anesthesias and operations.

[Study of spectrum processing method for Raman microscopy on single living cell].

To study the spectrum processing methods for Raman microscopy on single living cell and develop the pre-process techniques for Raman spectrum of single living cell to enhance the signal to noise ratio, sensitivity, and decrease the fluorescence influence, wiping off the cosmic rays was used to improve the spectrum. The spectra classification, spectra average and filtration were applied to enhance signal to noise ratio. The fluorescence was depressed for quantity analysis or utilized for analysis by comparing the background and the spectra. Results show that (1) comparing the spectra with short exposure time and more scans can wipe off the cosmic rays effectively. (2) the spectra classification, spectra average and filtration can improve the quality of spectra and can show some weak and sensitive bands. (3) sometimes the fluorescence has useful information. It is concluded that the proposed techniques for Raman spectrum of single cell in this paper can show the sensitive and weak intensity peaks and reflect the information of molecules structures very well.

Human red blood cell aging: correlative changes in surface charge and cell properties.

Red blood cells (RBCs) during microcirculation, aging and storage, lose N-acetylneuraminic acid (NANA) and other biomaterials thereby altering cell structures, some properties and functions. Such cell damage very likely underlies the serious adverse effects of blood transfusion. However, a controversy has remained since 1961-1977 as to whether with aging, the RBCs, suffering loss of NANA, do have a decreased charge density. Any correlation between the changes in the cell properties with cell aging is also not clear. Therefore, to remove the ambiguity and uncertainty, we carried out multiparameteric studies on Percoll fractions of blood of 38 volunteers (lightest-young-Y-RBCs, densest-old-O-RBCs, two middle fractions).We found that there were striking differences between the properties of Y-RBCs and O-RBCs. The ζ-potential of Y-RBCs decreased gradually with aging. Studies in parallel on RBC fractions incubated with both positively charged quantum dots and Sambucus Nigra-fluorescein isothiocyanate (FITC) along with their ζ-potentials provide for the first time direct visual evidence about the lesser amount of charge density and NANA on O-RBCs, and a collinear decrease in their respective ζ-potentials. Close correlation was found between the surface charge on an aging RBC and its structure and functions, from the cell morphology, the membrane deformability to the intracellular Hb structure and oxidation ability. This quantitative approach not only clarifies the picture but also has implications in biology and medicine.

Quantum dots as a sensor for quantitative visualization of surface charges on single living cells with nano-scale resolution.

We developed a technique using quantum dot (QD) as a sensor for quantitative visualization of the surface charge on biological cells with nano-scale resolution. The QD system was designed and synthesized using amino modified CdSe/ZnS nanoparticles. In a specially designed buffer solution, they are positively charged and can homogeneously disperse in the aqueous environment to label all the negative charges on the surfaces of living cells. Using a wide-field optical sectioning microscopy to achieve 2D/3D imaging of the QD-labeled cells, we determined the charge densities of different kinds of cells from normal to mutant ones. The information about the surface charge distribution is significant in evaluating the structure, function, biological behavior and even malignant transformation of cells.

[Qualitative analysis of Raman spectra based on pulse coupled neural network].

By studying on pulse coupled neural network (PCNN) and Raman spectra qualitative analysis, a method based on PCNN for Raman spectra qualitative analysis was proposed. After encoding the Raman spectra by using PCNN neurons' characteristics of fatigue and refractory period, the improved Horspool algorithm was used to match the code corresponding to the detected sample with all of the base code in the database one by one, and then their matching similarity was acquired to determine the sample type. Experimental results and analysis of data proved that the method proposed in this paper is accurate and effective for Raman spectra qualitative analysis. Meanwhile, traditional qualitative analysis method based on spectral template has some deficiencies, like that it is difficult to determine the characteristic peak of the detected sample and the matching analysis process has a high degree of redundancy. While our proposed method not only can avoid these deficiencies very well, but also needs a small amount of data storage. The requirement of the storage space was only 5.8% of that used in the traditional qualitative analysis method based on spectral template.

[Analysis of the effects of different medicines on hypercoagulability state variations of femoral shaft fracture and after its operation].

OBJECTIVE: To explore the role of Tong Mai Tang & Lornoxicam on the serum concentrations of tumor necrosis factor alpha (TNF-alpha), interleukin (IL-6) , D-dipolymer( D-Di), Platelet count (PLC) in treatment of femoral shaft fracture among period surgery time. METHODS: We selected 120 cases of traumatic femoral shaft fracture patients according to the inclusion criteria and exclusion criteria, which were randomized dividend into four groups (I, II, III, IV respectively) of the same size based on the random number table method of 30 patients each. Therapeutic methods of four groups following as: Group I, Tpanax Notoginseng Pills PO; Group II, Tpanax Notoginseng Pills PO, Lornoxicam For Injection, 8 mg IM; Group III, Tpanax Notoginseng Pills PO, Tong Mai Decoctions 200 mL PO; Group IV, Tpanax Notoginseng Pills PO, Lornoxicam For Injection 8 mg IM, Tong Mai Decoctions 200 mL PO. The above medications were administered to the four groups after admission to hospital the next day. Peripheral blood samples were taken for immune determination of pro-inflammatory cytokines of TNF-alpha, IL-6, D-Di, PLC in blood serum on the 2nd and 6th day before operation and on the 8th and 13th day after operation in the morning. And all patients received liver and kidney function examination 2nd and 13th day after admission. Analysis of variance and least significant difference-test were done with the help of SPSS 17.0 statistic software. RESULTS: The difference among four groups in TNF-alpha and IL-6 were significant (P < 0.05). And there were also significant statistic difference between group II/III/IV and group I group (P < 0.05). But the difference between group II and group III was insignificant (P > 0.05). However, the group contrast result between group IV and group II/III had statistics difference (P < 0.05). The difference in D-Di PLC at 6th day and 8th day were significant (P < 0.05). The group comparisons in group I/II/IV were also significant. There were non-statistics significance in group II compared 6th day/8th day with 2nd day (P > 0.05). The comparison between the 13th day with the first three time sections had statistics significance. And there were statistics significance at the 13th day between group IV and group II (P < 0.05). CONCLUSION: The serum concentrations of TNF-alpha, IL-6, D-Di and PLC level were significantly increased in peroperative period, These results seem to indicate that the Tong Mai Decoctions & Lornoxicam may play an important role in inhibiting the release of TNF-alpha, IL-6, D-Di and PLC into the blood stream and decreasing the incunabula complication at early traumatic stage. The Tong Mai Decoctions & Lornoxicam was the worth promoting screened China and the West union medication combination.

[Raman spectra quantitative analysis on materials with strong fluorescence background].

Aiming at the difficulty of Raman spectra quantitative analysis on materials with strong fluorescence background, together with baseline correction, a new normalization method was performed for concentration quantitative analysis of two kinds of solutions which exhibit strong fluorescence background, the methanol solutions with different concentrations and the mixed solutions of ethanol and methanol with different ratios of concentration. Meanwhile, the data fluctuation caused by collecting spectra data at different space-time was investigated by using statistical method of randomized blocks analysis of variance to evaluate the data fluctuation among different sample groups, and the function of our method proposed in this paper to eliminate this data fluctuation was discussed. Experimental results demonstrate that our proposed method can not only obtain satisfied accuracy for Raman spectra quantitative analysis on methanol with strong fluorescence background, with the mean relative error being 4.7%, but also effectively eliminate the data fluctuation among different sample groups, and the relative standard deviation of them was only 4.2%, indicating that it is possible to carry out simple, quick and precise quantitative determination of sample content for the materials with strong fluorescence background.

[Effects of Tongmai decoction on the perioperative changes of plasma TNF-alpha and IL-6 in patients with femoral fractures].

OBJECTIVE: To explore the effects of Tongmai decoction on the perioperative changes of serum concentrations of tumor necrosis factor alpha (TNF-alpha) and interleukin (IL)-6 in patients with femoral fractures, and conform the effectiveness of Tongmai decoction on inflammatory factors in patients with femoral fractures, providing the theoretical evidence for the clinical use of Tongmai decoction. METHODS: From October 2007 to May 2009, 60 patients with closed traumatic femoral fractures were selected according to the inclusion criterias and exclusion criterias. All the patients were randomly divided into three groups (group A, group B and group C). Twenty patients in group A (Tpanax Notoginseng pill group), 13 patients were male and 7 patients were female; ranging in age from 20 to 45 years, averaged 32.0 years; the disease course ranged from 2.0 to 26.0 h, with an average of 9.5 h. Twenty patients in group B (Tpanax Notoginseng pills and Lornoxicam injection group),12 patients were male and 8 patients were female; ranging in age from 23 to 42 years, averaged 31.0 years; the disease course ranged from 3.5 to 25.0 h, with an average of 13.6 h. Twenty patients in group C (Tpanax Notoginseng pill, Lornoxicam injection and Tongmai decoction group), 14 patients were male and 6 patients were female; ranging in age from 21 to 44 years, averaged 31.5 years; the disease course ranged from 4.6 to 29.0 h, with an average of 13.3 h. Among all the patients, 42 patients with fractures were fixed with femoral intramedullary nailing, and other 18 patients with femoral locking plate fixation. The patients in group A took Tpanax Notoginseng pills orally, 4 g each time and twice daily; the patients in group B took Tpanax Notoginseng pills orally as group A, and at the same time received intramuscular injection of Lornoxicam, 8 mg each time and once daily; the patients in group C took Tpanax Notoginseng pills orally and received intramuscular injection of Lornoxicam as group B, and at the same time took Tongmai decoction (R ) orally, 200 ml each time and twice daily. The above medications were administered to the three groups on the second day after admission to hospital. Peripheral blood samples were taken for determination of pro-inflammatory cytokines of TNF-alpha and IL-6 in blood serum on the 2nd and 6th days before operation and on the 8th and 13th days after operation. And all the patients were evaluated liver and kidney function at the 2nd and 7th days after admission. Analysis of variance and least significant difference-test were done with the help of SPSS 17.0 statistic software. RESULTS: The differences among three groups of TNF-alpha and IL-6 in blood serum at the 2nd day after admission and 2 days after operation had no statistical significance (P > 0.05). The TNF-alpha and IL-6 levels among 3 groups had statistical differences at the 7th day after admission and at the 7th day after operation (P < 0.05, P < 0.01). There were significant differences of TNF-alpha and IL-6 levels between the 7th day after admission and the 2nd day after admission, the 7th day after operation and the 2nd day after admission (P < 0.01). There were also significant differences of TNF-alpha and IL-6 levels between group C compared with group A and B at the 7th day after admission and the 7th day after operation(P < 0.05, P < 0.01). CONCLUSION: The serum concentrations of TNF-alpha and IL-6 level significantly increased in perioperative period. The results indicate that the Tongmai decoction may play an important role in inhibiting the release of TNF-alpha and IL-6 into the blood stream and decreasing the incunabula complication at early traumatic stage.

[Normalization methods for ethanol Raman spectra quantitative analysis].

By performing different Raman normalization methods for ethanol quantitative analysis, the authors proposed the method which used the highest band intensity of the ethanol Raman spectra in the maximum concentration as the normalization metric for ethanol concentration quantitative analysis. By using this method, the correlation coefficient was 0.999, the mean relative error was only 0.067 8, and the relative standard deviation (RSD) of the data from different experimental groups was 0.046 3. Both the validity and accuracy of this method were much better than the internal standard and ratio methods. Combined with baseline correction, the method can not only effectively resist the data fluctuation between different experimental groups, but also improve the accuracy of ethanol quantitative analysis obviously. The test using the method to measure the ethanol concentration of some wines from market proved that the relative standard deviations were all smaller than 0.012, indicating that the method is excellent for ethanol concentration quantitative analysis in commercial applications.

Effect of pH on molecular constitution and distribution of hemoglobin in living erythrocyte.

The molecular constitution of in situ hemoglobin (Hb) and their distribution in living erythrocyte were investigated versus pH using the technique of confocal Raman microscopy. Both Raman point spectra and line mapping measurements were performed on living erythrocytes in suspensions with pH values from 4.82 to 9.70. It was found that the Hb inside a living erythrocyte would dissociate into monomer/dimer when the cells are in low and high pH environments. In contrast to the homogeneous distribution of the Hbs in the cells in neutral suspension, there are more Hbs distributing around the cell membrane or binding to the membrane as pH increases. While in low pH, as the cell become spherical, most of the Hbs distribute to the central part of the cell. In summary, our investigation suggests that the variation of the external pH not only brings changes in the morphology and membrane structure of an erythrocyte, but also affects the constitution and distribution of its intracellular Hbs, thereby the flexibility of the cell membrane and the oxygenation ability of the Hb.

[Experimental study on the treatment of serious soft tissue injuries with strengthening the spleen and replenishing qi].

OBJECTIVE: To study the effects of Chinese drugs based on strengthening the spleen and replenishing qi treatment rule on neoformative capillaries and fibroblast during the soft tissue repair after serious trauma in rats, so as to explore the biological basis of the TCM theory "the spleen dominate extremities and muscles" applied to the treatment of soft tissue injuries. METHODS: The model rats were established by bleeding from femoral artery and lancing method, and the rats were randomly divided into the control group, strengthening the spleen group and activating blood and resolving stasis group. The samples were got from the tissue of the wounded area at the 5th, 10th and 15th days after oral administration of the traditional Chinese medicine. After fixation and section, the tissues were stained by CD31 and PCNA staining. The amount of the capillaries and fibroblasts in the tissue of the wounded area were observed through multi-purpose microscope (ZEISS Axioskop2). Quantitative analysis was carried out on Image-ProPlus image analyzer. RESULTS: The amount of the capillaries and fibroblasts in the wounded tissue in the strengthening the spleen group were larger than that in the control group at the 5th, 10th and 15th day. And the proliferation speed of capillaries and fibroblasts was faster than those in the control group or the activating blood and resolving stasis group. CONCLUSION: The Chinese drugs according to strengthening the spleen and replenishing qi treatment rule were effective to promote growth of the granulation tissue and facilitate healing of the wounded area. And it has better effect than the treatment of promoting blood circulation and removing stasis.

Confocal Raman microscopy on single living young and old erythrocytes.

Raman confocal microscopy, including the techniques of point Raman spectra, line mapping, 2D mapping, and time-dependent spectrum monitoring performed with 514.5 nm excitation light, was used in a comparative study on the distribution and oxidation states of hemoglobin (Hb) in young and old mature erythrocytes. It is demonstrated that in contrast to the homogeneous distribution of the Hb in young cells, there are more Hb distribution around the cell membrane in old erythrocyte. The proteins exhibit some extent of aggregation and conformational change, present less ability of oxidation, and lower oxygenation speed than the Hb in young erythrocytes. Our results also provide the first direct evidence of some intermediate oxygenated states of Hb between the two fully oxygenated (R) and deoxygenated (T) states in living erythrocyte, and give detail information about the conformational change of the intracellular Hb with time during the reoxygenation process.